CN111053716B

CN111053716B - Application of Fuzhuan tea extract in preparation of skin conditioning product

Info

Publication number: CN111053716B
Application number: CN201911306311.XA
Authority: CN
Inventors: 傅文燕; 丁敏; 胡适
Original assignee: Fengchao Medical Technology Shanghai Co ltd
Current assignee: Fengchao Medical Technology Shanghai Co ltd
Priority date: 2019-11-28
Filing date: 2019-12-18
Publication date: 2021-12-03
Anticipated expiration: 2039-12-18
Also published as: CN113694115A; WO2021104276A1; CN111053716A

Abstract

The invention discloses application of a Fuzhuan tea extract in preparing a skin conditioning product, wherein the skin conditioning product comprises a skin care product, a cosmetic, a nutriceutical and a dermatological application product for resisting acne, resisting oxidation, resisting inflammation, resisting stimulation, resisting aging or enhancing skin elasticity. The Fuzhuan tea extract of the present invention is intended to inhibit inflammation by specific action against Propionibacterium acnes and therefore can be used for preparing skin conditioning products.

Description

Application of Fuzhuan tea extract in preparation of skin conditioning product

Technical Field

The invention belongs to the technical field of medicines and health-care foods, and particularly relates to application of a Fuzhuan tea extract in preparation of a skin conditioning product.

Background

The Fu tea is a post-fermented tea which is prepared by using raw dark green tea or crude old green tea as a raw material and is reprocessed, belongs to one of dark green tea in six basic tea categories of China, and is also a fully fermented tea. Fu tea has a long history, and is called Fu tea because it is processed and prepared in summer overnight and has fragrance and action similar to that of Poria. The main form of the Fuzhuan tea is compressed tea, namely, in the manufacturing process, in order to facilitate transportation and storage, a producer compresses the Fuzhuan tea into a brick shape, so that the Fuzhuan tea looks like a brick, and is also named as the Fuzhuan tea. At present, the Fu tea is also sold in the forms of loose tea, cake tea and the like. Fu tea is mainly sold in areas of frontier regions such as Xinjiang, Gansu, Tibet and the like, is commonly called as 'side tea' or 'side selling tea', and is known as 'mystery tea on ancient silk roads' and 'black and gold on silk roads' in more than forty countries such as Zhongya, Western Asia and the like along the 'silk roads'.

Modern pharmacological research shows that the Fuzhuan tea has obvious health effects of promoting digestion, reducing blood fat, nourishing the stomach, losing weight, regulating carbohydrate metabolism, inhibiting tumor cells, enhancing immunity and the like. The tuckahoe tea can promote metabolism of human bodies, improve immunity, strengthen physique and delay senility after being drunk for a long time, and has obvious pharmacological health care and pathological prevention effects.

Research shows that the pharmacological activity of the Fuzhuan tea is the result of the combined action of the active ingredients of the tea and a dominant bacterium, namely Eurotium cristatum, which is naturally formed in the fermentation process. The eurotium cristatum plays an important role in the quality and the pharmacological effect of the Fu tea. Eurotium cristatum called as golden flower fungus in Fuzhuan tea, belongs to Eurotium cristatum of Tricholomataceae of Ascomycetales, and is suitable for growing in soil, Fuzhuan tea, Cordyceps, Chinese medicinal tablet and sawdust. Eurotium cristatum consists of ascocarp and hyphae. The eurotium cristatum can produce rich natural active ingredients, thereby endowing the Fuzhuan tea with unique color, fragrance and taste, and being the material basis of the Fuzhuan tea with the health-care effects of promoting digestion, reducing fat and losing weight, inhibiting tumor cells, treating cardiovascular diseases and the like. The eurotium cristatum also generates a large amount of special bioactive substances in the Fu tea in the process of co-fermentation of the Fu tea and the tea.

At present, there are many reports on the research of Fu tea. For example, the process of preparing instant black tea by liquid fermentation was reported by the lu courser (the master thesis of university of agriculture in Anhui, Eurotium cristatum, functional research thereof, and the director Yuepeng Xiang). Yuan Yong (thesis of Master academic thesis of Hunan agriculture university, the preliminary research on pharmacological efficacy and chemical components of Fuzhuan tea golden flower fungus extract, and the guiding teacher Liu Zhong Hua) reported the research on the extraction and separation of Eurotium cristatum spore powder. Eichhornia et al (Eichhornia, Liusu pure, Yuanqian, Zhao le, research on wall breaking method of Ascomyces guanfaciei ascospores, food and machinery 2011,27 (3): 137-. Lu et al (Advance Journal of Food Science and Technology 2016, 10 (8): 591-596.) reported the study of the active ingredients in Eurotium cristatum fermented black tea. Zou et al (Molecules 2014,19,17839-17847) reported alkaloid compounds produced by fermentation of Eurotium cristatum. Fu et al (Food Research International 2011, 44: 2999-3005.) report the mechanism of hypolipidemic activity of Fu tea. Yan et al (Food and Agricultural Immunology 2017, 28 (3): 388-402) report the effect and mechanism of Eurotium cristatum in Fuzhuan tea for activating immunity.

According to 26.09.2013, the original national quality inspection and quarantine bureau approves the geographical sign product protection of the Jingyang Fuzhuan tea, and the national quality supervision inspection and quarantine bureau announces that the quality inspection bureau approves the geographical sign product protection of the salt pond salt (river east salt), the salt pond black mud, the Jingyang Fuzhuan tea, the Yijun walnut and the Hongcuan wine (No. 137 in 2013) clearly requires the quality technology of the Fuzhuan tea, so that the subsequent research on the Fuzhuan tea is facilitated.

Disclosure of Invention

The invention aims to provide application of a Fuzhuan tea extract in preparing a skin conditioning product.

In order to achieve the purpose, the technical scheme adopted by the invention is as follows:

in a first aspect of the invention, the invention provides a use of an extract of Fuzhuan tea in the preparation of a skin conditioning product.

The skin conditioning products include anti-acne, anti-oxidant, anti-inflammatory, anti-irritant, anti-aging, or skin elasticity enhancing skin care products, cosmetics, nutraceuticals, dermatological use products that are anti-acne or hair follicle enhancing, anti-alopecia, skin elasticity enhancing, anti-aging, or wrinkle removing dermatological use products.

The Fuzhuan tea extract refers to a water-soluble and/or fat-soluble extract. In other embodiments of the present invention, the fu tea extract refers to supercritical CO₂Extract, organic solvent extract, aqueous extract or mixture thereof.

The organic solvent is ethanol.

The preparation method of the organic solvent extract comprises the following steps: grinding Fuzhuan tea into powder, wherein the diameter of the powder is not more than 0.5cm, adding 85% -95% ethanol into the Fuzhuan tea powder according to the solid-to-liquid ratio of 1:20(m/v), ultrasonically breaking the wall until no tea powder and thalli can be seen by naked eyes, then carrying out cold immersion for 18-28 hours, collecting filtrate, repeating for 2-5 times, carrying out reduced pressure distillation on the filtrate, concentrating the filtrate, drying at low temperature to obtain a crude extract, adsorbing and decoloring the crude extract by macroporous resin (AB-8 macroporous adsorption resin), eluting by 85% -95% ethanol, concentrating and drying by distillation, and carrying out low temperature freeze drying to obtain the organic solvent extract of the Fuzhuan tea.

The preparation method of the water extract comprises the following steps: grinding Fuzhuan tea into powder, wherein the diameter of the powder is not more than 0.5cm, adding water into the Fuzhuan tea powder according to the solid-to-liquid ratio of 1:10(m/v), uniformly stirring, ultrasonically breaking the wall until no tea powder and bacteria can be seen, standing for 2 hours at room temperature, and boiling for 2 hours in a boiling water bath to obtain a mixed solution; taking out the mixed solution, cooling, centrifuging at an ultra high speed, and taking the supernatant; filtering and sterilizing the supernatant with 0.22 μm filter membrane to obtain water extract of Fuzhuan tea. Concentrating and evaporating to dryness if necessary, and freeze-drying at low temperature.

The supercritical CO₂The preparation method of the extract comprises the following steps: grinding Fuzhuan tea into powder with diameter not more than 0.5cm, completely crushing tea powder and thallus, adding water vapor for pre-soaking until water content is increased to 40%, placing the pre-soaked Fuzhuan tea powder into an extraction tank, and continuously feeding CO into the tank₂Gradually extracting the crude extract with CO of the crude extract₂Sending to a cleaning tank, allowing the Fuzhuan tea extract to enter water phase, and passing through supercritical CO₂Extracting to obtain crude extract, hydrolyzing starch and fiber (cellulose, hemicellulose, etc.) with mixture of cellulase and alpha-amylase (concentration of 0.5% each), heating at 65 deg.C for 30 min to denature the enzyme, and centrifuging to remove impurities; removing residual protein (exudate) with 15kDa ultrafiltration column, bleaching extract with activated carbon and filtering and recovering, sterilizing the product with 0.2 μm filter membrane, and lyophilizing or packaging to obtain Fuzhuan tea supercritical CO₂And (3) extracting.

The Fuzhuan tea extract of the invention is intended to inhibit inflammation through specific action against Propionibacterium acnes, so as to be used as a skin conditioning product, in particular as an anti-inflammatory drug or cosmetic for resisting inflammation of the skin.

The present inventors have found that the fu tea extract has cosmetic and dermatological uses that have not been described previously.

The fu tea extract of the present invention is a cosmetic, nutraceutical, or dermatological composition for treating or preventing acne comprising the fu tea extract in combination with a suitable excipient. The composition may also comprise at least one anti-acne substance selected from sebum regulators, antibacterial and/or antifungal agents, keratolytic and/or keratolytic agents, astringents, anti-inflammatory and/or anti-irritant agents, antioxidant and/or free radical scavengers, cicatrizants, anti-ageing agents or moisturizers. The present invention also relates to cosmetic methods of applying the compositions of the present invention to an area of skin affected by the condition, or orally administering the compositions of the present invention to an individual with the condition.

The skin conditioning product may be selected from the group consisting of products that inhibit inflammatory mediators in keratinocytes, products that inhibit gene expression of matrix proteases, products that inhibit VEGF synthesis and release in keratinocytes, products that promote cellular collagen expression, products that increase cellular fibrillin synthesis, products that maintain stem cells dry, products that stimulate stem cell proliferation, products that promote ATP synthesis or enhance mitochondrial metabolic activity in hair follicle papilla fibroblasts, products that increase cell viability of hair follicles or reduce damage to hair follicle membranes, products that increase expression of the skin antimicrobial peptide hBD3 by removing and eliminating toxins, products that inhibit active oxygen production (products that prevent skin problems caused by dust, such as inflammation, etc.).

The skin conditioning product takes the Fuzhuan tea extract as the only active component, or a pharmaceutical composition containing the Fuzhuan tea extract.

The pharmaceutical composition containing the Fuzhuan tea extract is a pharmaceutical composition consisting of the Fuzhuan tea extract and one or more pharmaceutically-allowable auxiliary materials.

The content of the Fuzhuan tea extract in the skin conditioning product is 0.1-99 wt%.

The content of the Fuzhuan tea extract in the skin conditioning product is 0.5-90 wt%.

The auxiliary material is at least one of diluent, excipient, adhesive, filler, disintegrating agent, flavoring agent and sweetener.

The Fuzhuan tea extract can be prepared into a medicinal preparation with a conventional pharmaceutic adjuvant in pharmaceutics.

The pharmaceutical preparation is at least one of a capsule, a suspension, a tablet, a powder, an emulsion, a solution, a syrup or an injection.

The administration mode of the pharmaceutical preparation is oral administration and injection.

In the description of the present invention, the fu tea is preferably jing yang fu tea which meets the quality and technical requirements for fu tea as described in the bulletin of the quality inspection and inspection bureau of the national quality supervision, inspection and quarantine bureau of the ministry of health and safety on the geographic marking product protection of salt pond (large salt in river east), black mud in salt pond in city, jing yang fu brick tea, lijun walnut and hong chuan wine (No. 137 in 2013).

The invention also comprises a composition containing the Fuzhuan tea extract as an active ingredient according to the type of application. The composition comprises at least one pharmaceutically acceptable excipient, in particular a dermatologically acceptable excipient. If the composition is a cosmetic or dermatological composition, excipients suitable for topical application in vitro are used.

The composition of the invention may also comprise at least one pharmaceutical adjuvant known to the person skilled in the art, selected from thickeners, preservatives, fragrances, colorants, chemical or mineral sunscreens, humectants, thermal spring water and the like.

The composition of the present invention may further comprise at least one anti-acne compound selected from sebum regulators, antibacterial agents, antifungal agents, keratolytic agents, cuticle regulators, astringent agents, anti-inflammatory/anti-irritant agents, antioxidant/free radical scavengers, cicatrizants, anti-ageing agents and/or moisturizers.

The term "sebum regulator" for example means an inhibitor of 5-alpha-reductaseSuch as active substances

Zinc and its gluconate, salicylate and pyroglutamic acid salts also have sebum suppressive activity. Mention may also be made of spironolactone, an antiandrogen and an aldosterone antagonist, which significantly reduces the rate of sebum secretion after 12 weeks of application. Other extract molecules, such as seeds from Cucurbita pepo, pumpkin seed oil, and palmetto (palm cabbagage), limit sebum production by inhibiting 5-alpha-reductase transcription and activity. Other sebum regulators that affect sebum properties, such as linoleic acid, may also be used.

The terms "antibacterial agent" and "antifungal agent" refer to molecules that limit the growth or destroy pathogenic microorganisms, such as certain bacteria, such as propionibacterium acnes, or certain fungi (Malassezia furfur), the most traditional being preservatives commonly used in cosmetics or nutraceuticals, i.e. molecules with antibacterial activity (pseudopreservatives) such as caprylic acid derivatives (caprylylglycine, caprylin, etc.), such as hexylene glycol and sodium levulinate, zinc and copper derivatives (gluconate and PCA), phytosphingosine and its derivatives, benzoyl peroxide, piroctone olamine, zinc pyrithione, selenium disulfide, econazole, ketoconazole, or topical antibiotics such as erythromycin and clindamycin, etc.

The terms "keratolytic agent" and "keratolytic agent" refer to a substance that modulates or assists in the elimination of dead cells of the stratum corneum of the epidermis. Commonly used keratolytic/keratolytic agents include: the most commonly used keratolytic/keratolytic agents include: alpha-hydroxy acids (AHAS) of fruits (citric acid, glycolic acid, malic acid, lactic acid, etc.), AHA esters, combinations of AHAS with other molecules such as malic acid and almond protein

A combination of glycolic or lactic acid and arginine or a hydroxy acid and a lipid molecule such as

(lipid-hydroxy acids), amphoteric hydroxy acid complexes (AHCare), willow bark (white willow bark extract), azelaic acid and its salts and esters, salicylic acid and its derivatives such as octanoylsalicylic acid or in combination with other molecules such as salicylic acid and polysaccharides (beta-hydroxy acids or BHA), tazarotene, adapalene, and molecules of retinoids such as tretinoin, retinal, isotretinoin, and retinol.

The term "astringent" refers to a substance that helps narrow pores, most commonly used are polyphenols, zinc derivatives, and hamamelis virginiana.

The term "anti-inflammatory/anti-irritant" refers to a substance that limits the inflammatory response caused by cytokines or arachidonic acid metabolic mediators and has smooth and anti-irritant properties. Most conventional are glycyrrhetinic acid (glycyrrhiza derivative) and its salts and esters, alpha-bisabolol, Ginkgo biloba (Ginkgo biloba), marigold (calenula), lipoic acid, beta-carotene, vitamin B3 (niacinamide), vitamin E, vitamin C, vitamin B12, flavonoids (green tea, quercetin, etc.), lycopene or lutein, avocado sugar, avocado oil essence (avocado oleodestillate), arabinogalactan, lupin peptide, lupin total extract, quinoa peptide extract, cyclic ceramide

(oxazoline derivatives), anti-glycation agents such as carnosine, N-acetylcysteine, isoflavones such as genistein/genistin, daidzein/daidzin, spring water or spa water (eau d 'Avene), beauty care spring (eau de la Roche Posay), Saint Fang Wei (eau de Saint Gervais), You d' Uriage, Kerd (eau de Gamarde), Lycium barbarum extract, vegetarian amino acid peptides or compounds, topical dapsone, or anti-inflammatory agents.

The term "antioxidant/radical scavenger" refers to a molecule that reduces or prevents the oxidation of other chemicals. The antioxidant/radical scavenger that may be used in combination is preferably selected from the group consisting of: mercaptan and phenol, Glycyrrhrizae radix derivatives such as glycyrrhetinic acid and its salt and ester, alpha-bisabolol, semen Ginkgo extract, and herba Sidae Rhombifoliae (Calendu)la) extract, cyclic ceramide

(oxazoline derivatives), avocado peptides, trace elements such as copper, zinc and selenium, lipoic acid, vitamin B12, vitamin B3 (niacinamide, nigulamide), vitamin C, vitamin E, coenzyme Q10, krill oil (krill), glutathione, Butylated Hydroxytoluene (BHT), Butylated Hydroxyanisole (BHA), lycopene or lutein, beta-carotene, polyphenol families such as tannins, phenolic acids, anthocyanins, flavonoids such as green tea, red berries, cocoa, grapes, passion fruit (pasiflora incarnata) or Citrus (Citrus) extracts, or isoflavones such as genistein/genistin and daidzein/daidzin. The antioxidant group also includes anti-glycation agents such as carnosine or certain peptides, N-acetylcysteine, and antioxidant or free radical scavenging enzymes such as superoxide dismutase (SOD), catalase, glutathione peroxidase, thioredoxin reductase, and antagonists thereof.

Substances which can be used in combination for scar formation/repair barrier function are preferably vitamin a, panthenol (vitamin B5),

Lupeol, maca peptide extract, quinoa peptide extract, arabinogalactan, zinc oxide, magnesium, silicon, madecassic or asiatic acid, dextran sulfate, coenzyme Q10, glucosamine and derivatives thereof, chondroitin sulfate and total glycosaminoglycans (GAGs), dextran sulfate, ceramides, cholesterol, squalane, phospholipids, fermented or unfermented soybean peptides, plant peptides, marine polysaccharides, plant polysaccharides or biotechnological polysaccharides, such as algae extracts or fern extracts, trace elements, extracts of tannin-rich plants such as tannins derived from gallic acid, known as indigestible tannins or hydrolysable tannins, originally found in oak galls, and catechin tannins resulting from the polymerization of flavan units, models of which are provided by the catechu (Acacia catechu). The trace elements which may be used are preferably selected from the group consisting of copper, magnesium, manganese, chromium, selenium, silicon, zinc and mixtures thereofThe group (2).

Anti-aging agents which can be used in combination to treat acne in mature subjects are antioxidants, in particular vitamin C, vitamin A, retinol, retinal, hyaluronic acid of any molecular weight, alpha-linolenic acid, beta-linolenic acid, and beta-linolenic acid,

Lupin peptide and maca peptide extracts.

The most commonly used humectants/emollients are glycerin or its derivatives, urea, pyrrolidone carboxylic acid and its derivatives, hyaluronic acid of any molecular weight, glycosaminoglycans and any other polysaccharides of marine, vegetable or biotechnological origin, such as xanthan gum, and mixtures thereof,

Certain fatty acids such as lauric, myristic, monounsaturated and polyunsaturated omega-3, -6, -7 and-9 fatty acids (linoleic, palmitoleic, etc.), sunflower oil essence (sunflower oleodestillate), avocado peptide and cupuacu butter (cupuacu butter).

In some preferred embodiments of the invention, the extract of hoechuan tea of the invention may be used as a skin conditioning product in combination with a plant or animal unsaponifiable composition, for example, avocado and soybean unsaponifiables and unsaponifiable vegetable oils or animal oil concentrates, for example, sunflower oil or palm oil concentrates, or unsaponifiable-containing vegetable oils, for example, soybean oil and rapeseed oil, and unsaponifiable derivatives such as avocado furan, sterol esters and vitamin derivatives.

In some preferred embodiments of the present invention, the extract of Fuzhuan tea of the present invention may be used as a skin conditioning product in combination with avocado sugar (see application WO2005/115421), said composition being particularly suitable for the treatment of skin barrier repair and inflammation.

In some preferred embodiments of the invention, the Fuzhuan tea extract of the invention may be used as a skin conditioning product in combination with avocado peptides (see International application WO2005/105123), said composition being particularly suitable for the treatment of irritation and inflammation.

In some preferred embodiments of the present invention, the Fuzhuan tea extract of the present invention may be used as a skin conditioning product in a composition with avocado oil (see International applications WO2004/012496, WO2004/012752, WO2004/016106, WO 2007/057439).

In some preferred embodiments of the present invention, the extract of Fuzhuan tea of the present invention can be combined with

(avocado furan, which can be obtained by the process described in international application WO 01/21605) the composition is used as a skin conditioning product. The compositions are particularly suitable for the treatment of inflammation, the promotion of scarring and for their anti-ageing properties.

In some preferred embodiments of the present invention, the fu tea extract of the present invention may be used as a skin conditioning product in combination with avocado or soy unsaponifiables. The avocado and soybean unsaponifiables that may be used in combination are preferably mixtures of avocado furan unsaponifiables and soybean unsaponifiables in a ratio of about 1:3 to 2:3, respectively.

In some preferred embodiments of the present invention, the fu tea extract of the present invention can be used as a skin conditioning product in a composition with lupeol (FR2822821, FR 2857596). The compositions are particularly suitable for supporting scarring.

In some preferred embodiments of the present invention, the extract of Fuzhuan tea of the present invention may be used in combination with lupin peptides as a skin conditioning product, such as lupin peptides obtained according to the method described in application WO 2005/102259. The compositions are particularly suitable for the treatment of inflammation and are used for their anti-aging properties.

In some preferred embodiments of the present invention, the extract of Fuzhuan tea of the present invention may be combined with a total lupin extract (see International application WO2005/102259) for use as a skin conditioning product, said composition being particularly suitable for treating irritation.

In some preferred embodiments of the present invention, the fu tea extract of the present invention may be used in combination with lupin oil as a skin conditioning product, preferably sweet white lupin oil, such as sweet white lupin oil described in international application WO 98/47479.

In some preferred embodiments of the present invention, the extract of Fuzhuan tea of the present invention may be combined with an extract of maca peptides (see International application WO2004/112742) for use as a skin conditioning product. Said compositions are particularly preferred for their cicatrizing and anti-ageing properties.

In some preferred embodiments of the present invention, the Fuzhuan tea extract of the present invention may be used as a skin conditioning product in combination with rice peptide (see International application 2008/009709). The compositions are particularly preferred for their properties associated with stimulating melanogenesis and with melanin transfer.

In some preferred embodiments of the present invention, the extract of Fuzhuan tea of the present invention can be combined with circulating ceramide

(oxazoline derivatives) make up compositions useful as skin conditioning products, such as the cyclic ceramides described in International applications WO2004/050052, WO2004/050079 and WO 2004/112741. The compositions are particularly suitable for treating inflammatory reactions.

In some preferred embodiments of the present invention, the extract of fu tea of the present invention may be used in combination with an extract of quinoa as a skin conditioning product, in particular a peptide extract (see international application WO 2008/080974). The compositions are particularly suitable for the treatment of inflammatory diseases and skin barrier repair.

In some preferred embodiments of the present invention, the fu tea extract of the present invention may be used as a skin conditioning product in a composition with cupuacu butter (cupuacu butter). The compositions are particularly preferred for their wetting properties.

In some preferred embodiments of the present invention, the fu tea extract of the present invention may be used as a skin conditioning product in combination with rapeseed oil serum (rapeseed oleodesidate).

In some preferred embodiments of the present invention, the fu tea extract of the present invention may be used as a skin conditioning product in combination with corn oil serum (corn oleostillate).

All of these combinations, except the fu tea extract, comprise at least one other active compound, and may comprise two, three, four or more active compounds as described above.

In addition to these actives described above, alone or in combination with the actives mentioned above, the fu tea extract of the present invention may be used in combination with sunscreen actives known to those skilled in the art, such as UVB and/or UVA filters or shades or any inorganic and/or organic shades or filters, which will adjust their selection and their concentration according to the degree of protection sought.

As examples of sunscreen actives, mention may be made in particular of titanium dioxide, zinc oxide, methylenebis-benzotriazolyl tetramethylbutylphenol (brand name: TINOSORB M) and bis-ethylhexyloxyphenol methoxyphenyl triazine (brand name: TINOSORB S), octyl-salicylic acid, butylmethoxydibenzoylmethane, p-xylylene dicamphor sulfonic acid, 4-methylbenzylidene camphor, benzophenone, ethylhexyl methoxycinnamate, ethylhexyl dimethyl PABA and diethyl hexyl butamido triazone.

The fu tea extract of the present invention can be prepared with pharmaceutically or cosmetically, nutraceutically suitable carriers in a variety of formulations suitable for topical or oral administration.

According to a first variant, the various formulations are suitable for topical application and comprise creams, gels, emulsifiers, emulsions, balms, lotions, oils, aqueous or hydro-alcoholic or glycolic acid solutions, powders, patches, sprays or any other product for external application.

According to a second variation, a plurality of formulations are suitable for oral administration, wherein the fu tea extract may be included in a dietary supplement or a dietary composition.

In the context of the present invention, the dietary supplement may be provided in the form of hard or soft gelatin or vegetable capsules. The dietary supplement may thus comprise 10% to 100% by weight of fu tea extract.

Without limitation, the fu tea extract of the present invention may also be incorporated in dietary compositions such as foods, beverages, and nutraceuticals, including the following products:

1) dairy products such as cheese, butter, milk and other milk-like beverages, mixes and spreads containing milk products, ice cream and yogurt;

2) fat-based products such as margarines, spreads, mayonnaises, cooking fats, frying oils and vinaigrette;

3) cereal-based products consisting of cereals, such as bread and pasta products, whether or not these foods are cooked, baked or processed;

4) confections such as chocolate, confectionery, chewing gum, sweets, toppings (toppings), sorbet, cake coatings (icing) and other garnishes (garnishe);

5) alcoholic or nonalcoholic beverages, including soda water and other soft drinks, juices, dietary supplements, meal substitutes in the form of beverages, e.g. under the brand name Boost^TMAnd Ensure^TMThose meal substitutes sold, and;

6) various products such as eggs, processed foods such as soups, ready-to-use pasta sauces, prepared dishes and other products of the same type.

The fu tea extract can be incorporated into foods, nutraceuticals, dietary products, especially high protein products, or beverages directly and without further modification by means of techniques such as mixing, pouring, injecting, blending, adsorbing, kneading, and spraying.

The mode of administration and the dosing regimen and the optimal galenical form of the compounds and compositions of the invention can be determined on the basis of the criteria generally considered when establishing a pharmacotherapy, in particular a dermatological therapy, suitable for the patient, such as the age or weight of the patient, the severity of the overall condition of the patient, the tolerance to said therapy, the apparent side effects and the skin type.

The invention also relates to a method for the cosmetic treatment of acne, in particular the application of the cosmetic composition of the invention to the affected skin area.

The invention also relates to a method for the cosmetic treatment of acne by the oral route, in particular the oral administration of the nutraceutical composition of the invention to the affected individuals.

The invention also relates to the use of an extract of Fuzhuan tea for the preparation of a cosmetic, nutraceutical or dermatological composition for the treatment or prevention of acne.

In the context of cosmetic or dermatological use, the composition will preferably be formulated in a formulation suitable for topical application.

In the context of use in food, the composition will preferably be formulated in a formulation suitable for oral administration for nutritional or cosmetic purposes (beauty foods).

Preferably, the Fuzhuan tea extract also has specific effects of stimulating proliferation and maintaining the activity of stem cells, resisting aging and promoting healing. The Fuzhuan extract had the effect of increasing the expression of the ratio of ALDH positive cells, Ki67, on human epidermal stem cells as described in the examples below.

Preferably, the extract of Fuzhuan tea and the extract of Fuzhuan tea according to the present invention are intended to inhibit inflammation by specific action against Propionibacterium acnes. The fu tea extract specifically locks in the acne inflammatory process that plays a central role in lesion development and exacerbation. Thus, the examples presented below show that the fu tea extract has several effects on keratinocytes:

anti-inflammatory action: inhibiting inflammatory early mediators (IL-1 alpha, IL-1 beta) and late mediators (IL-8); inhibition of keratinocyte-induced MMPs (MMP-2 and MMP-9); inhibition of VEGF release.

The Fuzhuan tea extract also has the following effects on Propionibacterium acnes: inhibiting propionibacterium acnes from inducing IL-8; inhibiting Propionibacterium acnes induces MMP-9.

In some aspects of the invention, the invention also comprises the application of the Fuzhuan tea extract for maintaining and/or increasing the skin vitality and elasticity, reducing the skin injury, resisting aging and the like by local or oral administration. Such as maintenance/augmentation of skin cell type I collagen, resistance to light-induced skin damage and aging, etc., as described in the examples.

In another aspect of the present invention, the present invention also includes the use of said fu tea extract for topical or oral administration for enhancing hair follicles and reducing loss of skin appendages, preferably hair loss. For example, the examples describe maintaining and/or increasing collagen type V and/or fibrillin-1 expression and/or for increasing proliferation of skin fibroblasts, hair follicle fibroblasts and/or for increasing cell viability and/or ATP synthesis and/or mitochondrial activity and/or reducing cell damage and/or reducing cell senescence.

In one embodiment of the present invention, a cosmetic treatment method comprises topically applying the present fu tea extract or a composition comprising the same to all or part of the body selected from: legs, feet, axilla, hands, thighs, abdomen, neck line, neck, arms, torso, back, face, including skin appendages, preferably hair, and/or scalp, more advantageously scalp.

The term "aging" and the term "skin aging" in the context of the present invention are used to describe visible changes in the appearance of the skin and those detectable by touch, such as, for example, wrinkles, fine lines, roughness, expression lines, stretch marks, discontinuities, deep wrinkles, sagging of the skin (e.g., cheek sagging, under-eye puffiness, chin), increased pore size, loss of elasticity, loss of resiliency, loss of firmness, elastosis, abnormal differentiation, hyperkeratosis, keratosis, skin color changes (e.g., blotches, redness, or under-eye puffiness), formation of hyperpigmented regions (e.g., age spots, yellow spots, or freckles), loss of suppleness, cellulite skin, loss of collagen structure and formation of the stratum corneum, dermis, epidermis, vascular system (e.g., spider veins or telangiectasia), in a non-limiting sense, Or other histological changes of these tissues close to the skin. Skin aging is a process with two major components: chronological aging, which is attributed to the passage of time; and photo-induced aging, which is attributed to the level of exposure to ultraviolet radiation and is referred to as photo-aging. The sum of several environmental factors, such as exposure to tobacco smoke, exposure to pollution, and climatic conditions (e.g., cold and/or wind), also contribute to skin aging. In the context of the present invention, a "skin anti-aging treatment" is a treatment for preventing, delaying, and/or reducing aging of human skin.

Due to the adoption of the technical scheme, the invention has the following advantages and beneficial effects:

the fu tea extract of the present invention is intended to inhibit inflammation by a specific action against propionibacterium acnes, and has several actions against keratinocytes: the Fuzhuan tea extract achieves the pharmaceutical function of resisting acne by inhibiting the gene expression of inflammatory mediators or matrix protease in keratinocytes and inhibiting the synthesis and release of VEGF in the keratinocytes; the anti-aging function is realized by promoting the expression of cell collagen, increasing the synthesis of cell fibrillin, maintaining the dryness of stem cells and stimulating the proliferation of stem cells; the dermatological use of hair follicle enhancement and anti-hair loss is achieved by promoting ATP synthesis or enhancing mitochondrial metabolic activity in hair follicle papilla fibroblasts, increasing cell viability of hair microfollicles or reducing microfollicle membrane damage; the skin care product can prevent skin problems caused by Asian dust by removing and eliminating toxins, increasing the expression of the skin antimicrobial peptide hBD3 and inhibiting the generation of active oxygen, and provides a new path for skin care such as anti-aging skin care, acne resistance, alopecia prevention, dust influence prevention and the like.

Detailed Description

In order to more clearly illustrate the invention, the invention is further described below in connection with preferred embodiments. It is to be understood by persons skilled in the art that the following detailed description is illustrative and not restrictive, and is not to be taken as limiting the scope of the invention.

Example 1

The preparation method of the Fuzhuan tea extract comprises the following steps:

grinding Fuzhuan tea into powder with diameter not more than 0.5cm, completely crushing the Fuzhuan tea powder with a dry wall breaking machine, adding water vapor for pre-soaking until the water content is increased to 40%, putting the pre-soaked Fuzhuan tea powder into an extraction tank, and continuously extractingFeeding CO into the tank₂And the crude extract is gradually extracted. CO with crude extract₂Sending to a cleaning tank, allowing the Fuzhuan tea extract to enter water phase, and passing through supercritical CO₂Extracting to obtain crude extract, hydrolyzing starch and fiber (cellulose, hemicellulose, etc.) with mixture of cellulase and alpha-amylase (concentration of 0.5% each), heating at 65 deg.C for 30 min to denature the enzyme, and centrifuging to remove impurities; removing residual protein (collecting exudate) with 15kDa ultrafiltration column, bleaching extract with activated carbon and filtering and recovering, sterilizing the product with 0.2 μm filter membrane, and lyophilizing or packaging to obtain Fuzhuan tea supercritical CO₂And (3) extracting.

The Pu' er tea extract is prepared according to the same method and is used as a reference substance of subsequent experiments.

Example 2

Fuzhuan tea extract for enhancing type I collagen

The fuzhuan tea extract prepared in example 1 was added to a culture solution (DMEM culture solution) of human skin fibroblasts (ATCC) (final concentration of the fuzhuan tea extract in the culture solution was 0.5% by mass to volume), to confirm that the fuzhuan tea extract promotes type I collagen synthesis at a cellular level.

The collagen was measured by using PICP kit (procollagen type I C peptidase immunoassay kit, R)&D Systems) were used. Human skin fibroblasts were cultured in DMEM at 2X 10⁵The individual cells/well were aliquoted into 6-well plates. After cell adhesion was confirmed, the fu tea extract as an active ingredient was dissolved in a culture solution (the final concentration of the fu tea extract in the culture solution was 0.5% by mass/volume) to prepare a treatment group, the pu' er tea extract at the same concentration was prepared as a control group, and the culture solution was used alone in the blank group. At 37 deg.C, 5% CO₂The cells are cultured in an incubator under the conditions for 24 hours, the content of the type I collagen in the cell lysate is measured, the results are shown in table 1, and the results show that the poria cocos tea extract can effectively promote the expression of the collagen.

TABLE 1 collagen concentration

	Treatment group	Control group	Blank group
				Type I collagen concentration [ mu ] g/ml	0.030	0.002	0.002
Increase ratio (%)	1500	-	-

Example 3

Effect of Fuzhuan tea extract on skin model

This example was performed using the fu tea extract prepared in example 1.

Skin aging test by ultraviolet irradiation was carried out on the skin, and the collagen fiber network at the epidermal-dermal junction in the skin was damaged by ultraviolet irradiation. Thus, the amount and morphology of collagen fibrils serves as an indicator of skin aging.

The fu tea extract prepared in example 1 was dissolved in DMEM medium to give a final concentration of 0.5% by mass/volume as a treatment group; pu' er tea extract of the same concentration was used as a control group, and the same DMEM medium without any tea extract was used as a blank group. Before each irradiationThe skin surface of hairless mice was coated with 100. mu.l of Fuzhuan tea extract, and the ultraviolet irradiation test was carried out after 30 minutes. The method of ultraviolet irradiation is as follows: two 40W long-wave ultraviolet lamps and two 40W medium-wave ultraviolet lamps (from Philips, 320-400nm for long wave and 320-320 nm for medium wave) are adopted, and the distance between the lamps and the skin is 35 cm. Irradiation was carried out for 2 hours per day for 6 weeks. Detecting with ultraviolet intensity tester (photoelectric instrument factory of Beijing university of teachers and universities), and accumulating irradiation intensity to long wave 165J/cm²Medium wave ultraviolet 4.667J/cm². The animals were sacrificed at 6 weeks, the skin tissue of hairless mice receiving uv irradiation was stained by Masson's Trichrome, the stained area was observed by an optical microscope, the stained area of the control group was taken as a standard area of 100%, and the results of the stained area of the treatment group and the stained area of the control group are shown in table 2, and it can be seen from table 2 that the fu tea extract, i.e., the treated group, can significantly enhance the network area of collagen fibers compared to the control group, suggesting that the fu tea extract can be used for preparing anti-photoaging products.

TABLE 2

	Mean value of	SD	p value
				Blank group	100	4.5
Control group	102.61	7.11	p＞0.05
				Treatment group	164.51	14.7	p＜0.05

Artificial skin (EpiDermFT from Mattek, Ashland, USA)^TM) Stabilization was carried out in a special medium (EFT-400) for 18 hours. As a treatment group, 100 μ l of the fu tea extract prepared in example 1 was coated on the surface of artificial skin at a concentration of 0.5% (mass to volume, DMEM medium) as an effective ingredient; pu' er tea extract with the same concentration is used as a control group, and a blank DMEM culture medium is used as a blank group. After 30 minutes, the artificial skin was irradiated by a 40W medium-wave ultraviolet light source (from Philips) at a position 10cm below the source (irradiation time: 5min, cumulative dose: 30 mJ/cm)²) Then, conventional cultivation is performed. After 24 hours, the artificial skin tissue was fixed with 10% formaldehyde and embedded in paraffin, by Masson's Trichrome: M&T) and cut into 5 μm tissues, thereby observing and calculating the dyeing area of collagen fibers (collagen fibers), the results are shown in table 3, and the results show that the fu tea extract prepared in example 1 has an anti-aging effect under light conditions.

TABLE 3

	Mean value of	SD	p value
				Blank group	100	7.92
Control group	96.51	5.55	p＞0.05
				Treatment group	144.51	12.73	p＜0.05

Example 4

The effect experiment of preventing and treating acne was carried out on the fu tea extract prepared in example 1.

A keratinocyte monolayer model; research on expression of L-1 alpha, IL-1 beta, IL-8, MMP-2 and MMP-9, keratinocyte/Propionibacterium acnes co-culture model; study of MMP-9 expression, keratinocyte monolayer model; study of the effects of IL-1 α and VEGF, keratinocyte/Propionibacterium acnes co-culture model; model building, experimental methods, real-time RT-PCR methods and results calculation for the study of the effects of IL-8 production reference (CN 102481245B). The results are as follows:

fu tea extract for initial inflammatory mediators: effects of IL-1 α and IL-1 β genes encoding IL-1 α and IL-1 β were analyzed for expression.

After 48 hours of treatment, 0.5% and 1% fu tea extract were tested to significantly inhibit IL-1 α gene expression (as shown in table 4) and IL-1 β gene expression (as shown in table 5) in keratinocytes, with 1% pu' er tea extract as a control and the blank containing only culture medium.

Table 4: IL-1 alpha gene expression in keratinocytes

IL-1 alpha assay	RQ	Inhibition%	p value
				Blank space	1
1% Pu' er tea extract	0.95	-5	p＞0.05
				0.5% Fuzhuan tea extract	0.32	-68	p＜0.05
1% Fuzhuan tea extract	0.15	-85	p＜0.05

Table 5: IL-1 beta gene expression in keratinocytes

The results in tables 4 and 5 show that the Fuzhuan tea extract can effectively inhibit the expression of IL-1 alpha and IL-1 beta of keratinocytes and effectively resist inflammation.

Effect of fu tea extract on IL-8 expression: secondary inflammatory mediators

1. Analysis of expression of genes encoding IL-8

After 48 hours of treatment, 0.5% and 1% fu tea extract were tested to significantly inhibit IL-8 gene expression in keratinocytes (as shown in table 6).

Table 6: IL-8 gene expression in keratinocytes

IL-8 assay	RQ	Inhibition%	p value
				Blank space	1
1% Pu' er tea extract	0.96	4	p＞0.05
				0.5% Fuzhuan tea extract	0.21	-79	p＜0.05
1% Fuzhuan tea extract	0.12	-88	p＜0.05

As can be seen from the data in Table 6, the Fuzhuan tea extract can inhibit the expression of IL-8 gene in keratinocytes, and effectively inhibit the generation of inflammation.

2. Effects on IL-8 production in keratinocytes stimulated by Propionibacterium acnes.

Treatment of keratinocytes with a Propionibacterium acnes suspension strongly stimulated IL-8 release, a secondary marker of inflammation (as shown in Table 7). Pretreatment with the Fuzhuan extract for 60 minutes inhibited IL-8 release by keratinocytes, with inhibition of IL-8 production beginning at 0.5% and reaching completion at 5%.

Table 7: analysis of IL-8 in keratinocyte/Propionibacterium acnes cocultures

As can be seen from the data in Table 7, the Fuzhuan tea extract can effectively reduce the release of IL-8 mediated by Propionibacterium acnes, and has the effect of remarkably inhibiting inflammation mediated by the Propionibacterium acnes.

Effect of Fuzhuan tea extract on matriptase MMP-2 and MMP-9

Analysis of MMP-2 expression

Treatment of keratinocytes with 0.5% fu tea extract for 48 hours resulted in significant inhibition (-75% inhibition) of MMP-2 gene expression (as shown in table 8).

Table 8: MMP-2 gene expression in keratinocytes

MMP-2	RQ	Inhibition%	p value
				Blank space	1
0.5% Pu her tea extract	1.05		p＞0.05
				0.5% Fuzhuan tea extract	0.255	-75	p＜0.01

As can be seen from the data in Table 8, the Fuzhuan tea extract can effectively reduce the gene expression of keratinocyte MMP-2 and has the effect of remarkably inhibiting inflammation diffusion.

Analysis of MMP-9 expression

Among the matrix proteases involved in the pathology of acne, MMP-9 plays a particularly important role. In fact, this protease is up-regulated by proinflammatory cytokines, and also by propionibacterium acnes.

As shown in Table 9, analysis of the Q-PCR results showed: MMP-9 gene expression was inhibited in keratinocytes after 48 hours of treatment with the fu tea extract (at 0.5% -45% inhibition and at 1% -69% inhibition).

Table 9: MMP-9 gene expression in keratinocytes

MMP-9	RQ	Inhibition%	p value
				Blank space	1
1% Pu' er tea extract	1.05		p＞0.05
				0.5% Fuzhuan tea extract	0.55	-45	p＜0.01
1% Fuzhuan tea extract	0.31	-69	p＜0.01

As can be seen from the data in Table 9, the Fuzhuan tea extract can effectively reduce the gene expression of keratinocyte MMP-9, and has the effect of remarkably inhibiting inflammation diffusion.

Similarly, modulation of MMP-9 expression was also assessed in a keratinocyte/Propionibacterium acnes co-culture model (as shown in Table 10). Using this model, the stimulatory effect of Propionibacterium acnes bacterial suspension on MMP-9 gene expression (+ 55%) was demonstrated. In contrast, pretreatment of keratinocytes with fu tea extract resisted and also inhibited this stimulatory effect against propionibacterium acnes (51% and 67% relative to keratinocytes treated with bacterial suspension).

Table 10: MMP-9 gene expression in keratinocyte/Propionibacterium acnes coculture

As can be seen from the data in Table 10, the Fuzhuan tea extract can effectively reduce the gene expression of MMP-9 mediated by Propionibacterium acnes, and has the effect of remarkably inhibiting inflammation mediated by Propionibacterium acnes.

Effect of Fuzhuan tea extract on angiogenesis (VEGF)

The potential inhibitory effect of the fu tea extract on the synthesis and release of VEGF by keratinocytes was studied and the results are presented in table 11. PMA stimulation of keratinocytes for 24 hours induced a large and significant release of VEGF (+ 266%). In contrast, pretreatment of keratinocytes with fu tea extract for 24 hours was on this effect and significantly inhibited this effect.

Table 11: analysis of VEGF in keratinocytes

As can be seen from the data in table 11, the fu tea extract can effectively reduce the secretion of VEGF, and has the effect of significantly inhibiting local vascular inflammation.

Example 5

Example 1 Fuzhuan tea extract prepared enhances collagen V expression in hair follicle fibroblasts

Normal human fibroblasts were cultured in FGM medium (Lonza) supplemented with Fuzhuan tea extract at final concentration of 0.5% and 1% by mass/volume for 48 hours at 37 deg.C under 5% CO₂And then the cell culture medium is removed. The FGM medium of Pu' er tea extract at a mass-to-volume ratio of 1% was used as a control, and the same FGM medium without any tea extract was used as a blank control. The cell layer was lysed with ammonium hydroxide lysate (manufactured by MIBio corporation) at 4 ℃ for 10 minutes, and then collagen type V was measured with an anti-collagen V antibody diluted to 1/4000 in a buffer solution (PBS). After 60 minutes, the secondary antibody diluted to 1/25000 was applied for 60 minutes. After washing, ECL revealing solution (manufactured by seimerfin) was added and the fluorescence was measured (ENVision, PerkinElmer). The fluorescence results were normalized to the fluorescence obtained in the control medium (control) and compared to the fluorescence obtained in each conditionThe results obtained for the correlation of the amounts of collagen obtained were the average (n: 3) (SD: standard deviation) of 3 measurements, and the results are shown in table 12.

TABLE 12 Hair follicle fibroblast type V collagen expression

And (4) conclusion: in the analyzed hair follicle-derived fibroblasts, the Fuzhuan extract increases the collagen V expression by at least 34% and 104%, and shows that the Fuzhuan extract can remarkably enhance the property of the collagen expression, and can be used in medicines, cosmetics and nutraceuticals for enhancing hair follicles and reducing alopecia.

Example 6

Fuzhuan tea extract for increasing synthesis of fibrillin-1

Culturing normal human fibroblast cells in FGM medium (Lonza) at 37 deg.C under 5% CO for 48 hr₂. The fu tea extract prepared in example 1 as an active ingredient was dissolved in the culture solution (final concentration of fu tea extract in the culture solution was 0.5% and 1% by mass/volume as a treatment group, 1% by mass/volume of pu' er tea extract as a control group, and the blank group had only the culture solution). At 37 deg.C, 5% CO₂The culture was continued in the incubator under the conditions for 48 hours, and fibroblasts were collected and then lysed with a lysis buffer (produced by Biyuntian Co.) at 4 ℃ for 30 minutes to perform immunoblotting (Western Blots), and the protein concentration was measured by the BCA method. All samples were normalized to the total amount of Protein (n ═ 3), and quantitative studies were performed using a chemiluminescent substrate using an anti-fibrillin-1 antibody (LS-BIO) and a secondary antibody that was a peroxidase-conjugated antibody (Protein Simple), with the results shown in table 13.

TABLE 13 fibrillin-1 expression

Group of	Mean value of	SD	p value
				Blank [ Medium Only ]]	100	6
Pu' er tea extract 1% (w/v culture medium)	102.5	11.1	p＞0.05
				Fuzhuan tea extract 0.5% (w/v culture medium)	165.6	15.5	p＜0.05
Fuzhuan tea extract 1% (w/v culture medium)	205.4	25.2	p＜0.05

And (4) conclusion: the fu tea extract prepared in example 1 of the present invention showed increased synthesis of fibrillin-1 in fibroblasts compared to the control.

Example 7

Fuzhuan tea extract for promoting ATP synthesis increase in hair follicle papilla fibroblasts

Non-pathological human fibroblasts from hair follicle papilla were cultured in DMEM medium for 24 hours, and then the fu tea extract prepared in example 1 as an effective ingredient was dissolved in the culture solution (final concentration of fu tea extract in the culture solution was 0.5% and 1% by mass/volume ratio as a treatment group, 1% by mass/volume ratio of pu' er tea extract as a control group, and the blank group had only the culture solution). At 37 deg.C, 5% CO₂The cultivation is continued for 6 days in the incubator with the condition, and the final concentration of the Fuzhuan tea extract is 0.5 percent and 1 percent by mass-volume ratio. ATP levels were measured by enzymatic methods (luciferin/luciferase complex; ATP Bioluminescence kit Roche Diagnostics) after 6 days (as shown in Table 14).

TABLE 14 ATP content

And (4) conclusion: the fu tea extract showed increased ATP synthesis compared to the control, confirming its use for enhancing the activity of hair follicles.

Example 8

Example 1 Fuzhuan tea extract prepared to enhance mitochondrial metabolic activity of hair follicle papilla fibroblasts

Culturing non-pathological human fibroblast of hair follicle papilla in DMEM medium at 37 deg.C and 5% CO for 24 hr₂. The fu tea extract prepared in example 1 as an active ingredient was dissolved in the culture solution (final concentration of fu tea extract in the culture solution was 0.5% and 1% by mass/volume as a treatment group, 1% by mass/volume of pu' er tea extract as a control group, and the blank group had only the culture solution). At 37 deg.C, 5% CO₂The culture was continued for 6 days in the conditioned incubator. By MTT (3- [4, 5-dimethylthiazol-2-yl) in the presence of succinic dehydrogenase]-2, 5-diphenyltetrazolium bromide) was measured for mitochondrial activity of fibroblasts. The resulting precipitate was extracted with DMSO, and the optical density of the DMSO solution was measured at 540nm (as shown in table 15).

TABLE 15 relative mitochondrial Activity

	Mean value of	SD	p value
				Blank [ Medium Only ]]	100	14.3
Pu' er tea extract 1% (w/v culture medium)	91.3	11.3	p＞0.05
				Fuzhuan tea extract 0.5% (w/v culture medium)	138.4	20.5	p＜0.05
Fuzhuan tea extract 1% (w/v culture medium)	176.5	25.5	p＜0.05

And (4) conclusion: the fu tea extract prepared in example 1 of the present invention showed an increase in mitochondrial activity of hair follicle papilla fibroblasts, which is involved in the enhancement of hair follicles, compared to the control.

Example 9

Example 1 Effect of Fuzhuan tea extract on micro hair follicles

The method for reconstructing the Fuzhuan tea extract comprises the following steps: the micro-follicle model involves co-culturing papillary fibroblasts, keratinocytes from the outer sheath of the hair follicle, and melanocytes in three dimensions, which constitute a reconstructed organ model closest to the hair follicle due to the ability to integrate the neuro-epidermal-mesenchymal interactions between the various cell types.

Non-pathological papilla fibroblasts were cultured in DMEM medium for 3 days to form monolayer epidermal papilla. Then, melanocytes and keratinocytes of the outer skin of hair follicles were added to the above-mentioned single-layered epidermal papilla to form micro hair follicles; after 24 hours of culture, the Fuzhuan tea extract prepared in example 1 as an effective component was dissolved in a culture solution (the final concentration of the Fuzhuan tea extract in the culture solution is 0.5% by mass/volume as a treatment group, the Pu' er tea extract in the final concentration of 0.5% by mass/volume as a control group, and the blank group is only the culture solution.) at 37 ℃ and 5% CO₂The incubation was continued in the conditioned incubator for 48 hours. The treatment was carried out for 48 hours, and the medium and the microcapillaries were sampled for analysis.

The viability of the microcapillaries was measured by the PrestoBlue method (Thermo Fisher Scientific) and determined 48 hours after treatment, with the values in table 16 expressed as the mean% of the controls normalized.

TABLE 16 relative cell viability

	Mean value of	SD	p value
				Blank [ Medium Only ]]	100	9.5
Pu' er tea extract 0.5% (w/v culture medium)	97.2	3.3	p＞0.05
				Fuzhuan tea extract 0.5% (w/v culture medium)	164.5	21.5	p＜0.05

And (4) conclusion: the fu tea extract prepared in the embodiment 1 of the invention has the capability of increasing the cell viability of the micro hair follicle, so that the fu tea extract has activity on enhancing hair follicle, and therefore, the fu tea extract can be used for preparing the medicine or the cosmetic for preventing alopecia.

Assessment of reduction of Membrane Damage at the level of micro Hair follicles

Measuring cell damage by colorimetry using lactate dehydrogenase assay, enablingCell damage can be quantified based on the measurement of lactate dehydrogenase activity in damaged cells in the culture medium. The increase in cell membrane damage and cell lysis leads to an increase in lactate dehydrogenase activity, which is proportional to the number of lysed cells. Non-pathological papilla fibroblasts were cultured in DMEM medium for 3 days to form monolayer epidermal papilla. Then, melanocytes and keratinocytes of the outer skin of hair follicles are added to the above-mentioned single-layer epidermal papilla to form micro hair follicles; after 24 hours of culture, the Fuzhuan tea extract prepared in example 1 as an effective component was dissolved in a culture solution (the final concentration of the Fuzhuan tea extract in the culture solution is 0.5% by mass and volume as a treatment group, 0.5% by mass and volume of Pu' er tea extract as a control group, and the blank group is only the culture solution.) at 37 ℃ and 5% CO₂The incubation was continued for 48 hours in the incubator under the conditions, and then the lactate dehydrogenase activity in the medium was measured using a lactate dehydrogenase release assay kit (produced in Biyun sky) (as shown in Table 17).

TABLE 17 relative cellular injury

	Mean value of	SD	p value
				Blank [ Medium Only ]]	100	12.5
Pu' er tea extract 0.5% (w/v culture medium)	93	6.1	p＞0.05
				Fuzhuan tea extract 0.5% (w/v culture medium)	35	10.5	p＜0.05

And (4) conclusion: the fu tea extract prepared in the embodiment 1 of the invention has the capability of relieving cell membrane damage, so that the fu tea extract has activity on strengthening hair follicles, and strengthening hair follicles and promoting the reduction of alopecia, namely, the fu tea extract prepared in the embodiment 1 of the invention can be used as a medicine or a cosmetic for preventing alopecia.

Example 10 Fuzhuan tea extract maintains Stem cell desiccating and proliferative Activity

This example was performed using the fu tea extract prepared in example 1.

Human epidermal stem cells (ESC, ATCC) at 2X 10⁵The individual cells/well were aliquoted into 6-well plates in DMEM medium. After confirming cell adhesion, the fu tea extract prepared in example 1 as an active ingredient was dissolved in a culture medium (final concentration of fu tea extract in the culture medium was 0.5% by mass/volume as a treatment group, 0.5% by mass/volume of pu' er tea extract as a control group, and only culture medium was used in a blank group) at 37 ℃ with 5% CO₂The incubation was continued for 24 hours in the conditioned incubator. Flow cytometry was used to determine the ALDH-and Ki 67-positivity of ESC cells, and the results are shown in Table 18.

TABLE 18 ALDH positivity and Ki67 positivity

	Treatment group	Control group	Blank group	p-value (treatment vs. control)
					ALDH positive rate	58.5％	29.1％	26.7％	p＜0.05
Ki67 positive rate	45.5％	15.5％	18.3％	p＜0.05

The results show that the Fuzhuan tea extract has very strong effects of maintaining dryness of the epidermal stem cells and stimulating the proliferation of the epidermal stem cells.

Example 11

The Fuzhuan tea extract prepared in example 1 copes with active oxygen production in human keratinocytes caused by Asian dust

Asian dust is a climatic phenomenon originating in deserts of China, Mongolia, China, Russia and the like. In these places, sand or yellow sand dust blows east with the wind and falls to the ground. In the north, such as Beijing, is affected by Asian dust almost every year. Asian dust contains organic and inorganic substances, and its physical properties and composition are different, even containing metals that may have biological effects. Large particles contained in Asian dust remain in place or around it, while particles having a diameter of 10 μm or less (particulate matter 10; PM10), and PM2.5, which has been a continuous outbreak in recent years, is also a part of this type of climate pollution.

It has been reported that the particles, when inhaled, can reach deep into the gas exchange region of the sub-segmental bronchi and lungs and may damage the respiratory system (Yanagisawa et al, Exp Biol Med 232: 1109-. A number of reports have been made of the health effects of asian dust on human respiratory disease. In experimental studies in mice, an increased oxidative stress-related index was found in the lungs of mice perfused with Asian dust (Naota et al, Toxicol Pathol.2010 Sep30. clinically, if the skin is directly exposed to Asian dust, it may cause contact dermatitis by allergic reactions. furthermore, dry and intense Asian dust storms are known to cause dry skin, keratinization, itching, acne, atopy by depriving the skin of moisture.

Culturing human epidermal new keratinocyte cells for 24 hr in DMEM at 37 deg.C and 5% CO₂. The Fuzhuan tea extract prepared in example 1 as an effective component was dissolved in a culture solution (final concentration of the Fuzhuan tea extract in the culture solution was 0.5% and 1% by mass/volume as a treatment group, 1% by mass/volume of Pu' er tea extract was used as a control group, and only the culture solution was used in a blank group), and then 30. mu.g/mL Asian dust samples were added respectively, and 5% CO was added at 37 ℃₂The incubation was continued for 24 hours in the incubator under the conditions. Asian dust samples were collected during the asian dust season on the balcony roof of a national trade third generation a building (beijing city). Human epidermal neonatal keratinocytes were washed twice with 10mL Phosphate Buffered Saline (PBS) and 1 min after addition of 1mM luminol and 20 units/mL horseradish peroxidase (HRP) using a Glomax20/20 luminometerChemiluminescence was measured in 2mL PBS and the results are shown in table 19.

TABLE 19 relative active oxygen levels

The results show that the Fuzhuan tea extract prepared in example 1 of the present invention can inhibit the generation of active oxygen in normal human keratinocytes mediated by Asian dust.

Example 12

Fuzhuan tea extract prepared in example 1 has effects of toxin detoxification and elimination factor and antimicrobial peptide expression

In the same manner as in example 11, human epidermal neonatal keratinocyte cells were cultured in DMEM medium at 37 ℃ and 5% CO for 24 hours₂. The Fuzhuan tea extract prepared in example 1 as an effective component was dissolved in a culture solution (final concentration of the Fuzhuan tea extract in the culture solution was 0.5% and 1% by mass/volume as a treatment group, 1% by mass/volume of Pu' er tea extract was a control group, and a blank group had only the culture solution), and then 30. mu.g/mL Asian dust samples were added respectively, and 5% CO was added at 37 ℃₂The incubation was continued for 24 hours in the conditioned incubator. Asian dust samples were collected during the asian dust season on the balcony roof of a state trade third generation a building (beijing city). Experiment each group of cells was washed twice with 10mL of phosphate buffer and total rna (total rna) was isolated from the cells using trizol (invitrogen). The isolated RNA was again purified using the RNA kit (Qiagen) and the quality of the RNA was analyzed using the Bioanalyzer2100 (Agilent). cDNA was synthesized from the isolated RNA using Superscript Reverse Transcriptase (RT) II kit, and quantitative analysis of the cDNA was performed by real-time reverse transcriptase polymerase chain reaction (qPCR). Use ofThe TaqMan gene expression assay kit (TaqMan gene expression assay kit, Applied Biosystems) detects the expression levels of GSTT1(Hs00184475_ m1), NQO1(Hs01045994_ m1) and hBD3(Hs00218678_ m1), and the results are shown in tables 20 to 22.

TABLE 20 NQO1 relative expression levels

	Average (%)	SD	p value
				No Asian dust was added	140.56	18.56
Blank control	100	11.27
				Pu' er tea extract 1% (w/v culture medium)	106.12	8.67	p＞0.05
Fuzhuan tea extract 0.5% (w/v culture medium)	266.36	30.15	p＜0.05
				Fuzhuan tea extract 1% (w/v culture medium)	487.36	45.22	p＜0.05

TABLE 21 GSTT1 relative expression levels

TABLE 22 hBD3 relative expression levels

	Average (%)	SD	p value
				No Asian dust was added	256.75	30.11
Blank control	100.00	12.33
				Pu' er tea extract 1% (w/v culture medium)	93.35	10.17	p＞0.05
Fuzhuan tea extract 0.5% (w/v culture medium)	197.69	15.98	p＜0.05
				Fuzhuan tea extract 1% (w/v culture medium)	499.26	50.76	p＜0.05

The results show that the Fuzhuan tea extract prepared in example 1 of the invention can increase the expression of the detoxin protein NQO1, the skin antimicrobial peptide hBD3 of natural immune factors and the GSTT1 of toxin-removing enzyme in normal human keratinocytes.

Example 13

Example 1 Fuzhuan tea extract prepared in example 1 has effect on anti-oxidized protein gene expression

In the same manner as in example 11, human epidermal neonatal keratinocyte cells were cultured in DMEM medium at 37 ℃ and 5% CO for 24 hours₂. The fu tea extract prepared in example 1 as an active ingredient was dissolved in a culture solution (final concentration of fu tea extract in the culture solution)The mass ratio of the extract to the extract was 0.5% and 1% by volume, the mass ratio of the extract to the extract. ) Then, 30. mu.g/mL Asian dust samples were added separately at 37 ℃ with 5% CO₂The incubation was continued for 24 hours in the conditioned incubator. Asian dust samples were collected during the asian dust season on the balcony roof of a state trade third generation a building (beijing city). Experiment each group of cells was washed twice with 10mL of phosphate buffer and total rna (total rna) was isolated from the cells using trizol (invitrogen). The isolated RNA was again purified using the RNA kit (Qiagen) and the quality of the RNA was analyzed using the Bioanalyzer2100 (Agilent). cDNA was synthesized from the isolated RNA using Superscript Reverse Transcriptase (RT) II kit, and quantitative analysis of the cDNA was performed by real-time reverse transcriptase polymerase chain reaction (qPCR). The results of measuring the expression levels of GPX1(Hs00829989_ gH), catalase (Hs 0015638 _ m1), TRX1(Hs01555212_ g1) and PRDX1(Hs00602020_ mH) using TaqMan gene expression assay kit (TaqMan gene expression assay kit, A pplied Biosystems) are shown in tables 23 to 26.

TABLE 23 relative expression levels of GPX1

	Average (%)	SD	p value
				No Asian dust was added	165.66	18.66
Blank control	100.00	7.15
				Pu' er tea extract 1% (w/v culture medium)	109.12	6.18	p＞0.05
Fuzhuan tea extract 0.5% (w/v culture medium)	198.22	210.35	p＜0.05
				Fuzhuan tea extract 1% (w/v culture medium)	455.33	50.62	p＜0.05

TABLE 24 relative catalase expression levels

	Average (%)	SD	p value
				No Asian dust was added	256.56	30.11
Blank control	100.00	16.67
				Pu' er tea extract 1% (w/v culture medium)	93.31	10.81	p＞0.05
Fuzhuan tea extract 0.5% (w/v culture medium)	194.13	20.74	p＜0.05
				Fuzhuan tea extract 1% (w/v culture medium)	398.44	41.94	p＜0.05

TABLE 25 TRX1 relative expression levels

	Average (%)	SD	p value
				No Asian dust was added	164.18	19.65
Blank control	100.00	18.66
				Pu' er tea extract 1% (w/v culture medium)	115.91	8.94	p＞0.05
Fuzhuan tea extract 0.5% (w/v culture medium)	208.81	22.84	p＜0.05
				Fuzhuan tea extract 1% (w/v culture medium)	299.32	35.31	p＜0.05

TABLE 26 relative expression levels of PRDX1

	Average (%)	SD	p value
				No Asian dust was added	301.57	32.66
Blank control	100.00	9.91
				Pu' er tea extract 1% (w/v culture medium)	123.31	5.57	p＞0.05
Fuzhuan tea extract 0.5% (w/v culture medium)	291.54	18.85	p＜0.05
				Fuzhuan tea extract 1% (w/v culture medium)	466.69	50.02	p＜0.05

The results show that the Fuzhuan tea extract prepared in example 1 of the invention can increase the relative expression levels of antioxidant enzymes GPX1, catalase, TRX1 and PRDX1 for scavenging active oxygen, and can effectively resist Asian dust mediated oxidative stress reaction.

Example 14

Example 1 anti-aging skin care assay of Fuzhuan tea extract

According to the mass percentage, 4% of the Fuzhuan tea extract prepared in the embodiment 1 of the invention, 5% of butanediol, 0.2% of nipagin ester and 90.8% of water are respectively weighed, and the raw material components are mixed to obtain the anti-aging skin lotion.

The prepared anti-aging skin care lotion is further detected, and the result is as follows: the anti-aging skin care lotion is a brownish red transparent solution, has fragrant and pleasant smell, and has very fresh and cool feeling and good skin feeling after being sprayed on the surface of the skin.

After the anti-aging skin lotion is stored for 3 years on a shelf at normal temperature, the anti-aging skin lotion does not deteriorate, and all sensory indexes, physical and chemical indexes and sanitary indexes of the product meet the corresponding national standards; the prepared anti-aging skin lotion is subjected to a heat resistance test for 24 hours at the temperature of 40 +/-1 ℃, then the skin lotion is recovered to the room temperature, the test shows that the skin lotion is not turbid after the skin lotion is recovered to the room temperature, and all sensory indexes, physical and chemical indexes and sanitary indexes of the skin lotion after the skin lotion is recovered to the room temperature meet the corresponding national standards. The prepared anti-aging skin care water is subjected to a cold resistance test for 24 hours at the temperature of minus 15 +/-1 ℃, then the skin care water is recovered to the room temperature, the detection shows that the skin care water is not turbid after the skin care water is recovered to the room temperature, and all sensory indexes, physical and chemical indexes and sanitary indexes of the skin care water after the skin care water is recovered to the room temperature meet the corresponding national standards.

24 volunteers were selected and randomized for gender. The anti-aging skin lotion prepared in the example was randomly applied to one of the left cheek and the right cheek and one of the left corner and the right corner of the eye of each volunteer, respectively, and used as an experimental group; one cheek and canthus of volunteers who did not use the anti-aging skin lotion prepared according to the present invention were used as a control group.

The volunteers respectively apply the anti-aging skin lotion prepared by the embodiment once in the morning and at night every day, the service cycle is 6 weeks, and other cosmetics cannot be used in the test cycle. Then, the anti-aging skin lotion prepared in this example was comparatively analyzed for the improvement of skin elasticity and wrinkles of volunteers using a skin elasticity tester (Bareiss) and a facial wrinkle imager (Visia), and the results are shown in table 27:

watch 27

Detecting items	Anti-aging skin care lotion
		Wrinkle (wrinkle)	Compared with the control group, the wrinkle depth of the experimental group becomes shallow, and the effect is very obvious
Elasticity of skin	Compared with the control group, the skin elasticity of the experimental group is obviously improved

The result shows that the anti-aging skin lotion prepared by the embodiment obviously has the effects of resisting wrinkles and enhancing skin elasticity.

Example 15

Anti-aging skin care lotion prepared from Fuzhuan tea extract prepared in example 1

According to the mass percentage, the camellia sinensis extract is 3% of the poria cocos tea extract prepared in the embodiment 1 of the invention, wherein the caprylic/capric triglyceride is 10%, the white oil is 5%, the stearyl alcohol is 2%, the glycerin is 3%, the xanthan gum is 0.5%, the sorbitan stearate is 2.5%, the sucrose cocoate is 2.5%, the citric acid monohydrate is 1%, the paraben is 0.5%, the citronellol is 0.1%, and the water is 69.9%. Respectively weighing the raw materials of the components, and then preparing the anti-aging skin care emulsion according to the following steps:

1) adding xanthan gum, glycerol, sorbitan stearate and sucrose cocoate into a water phase pot, uniformly stirring, and heating to 80-85 ℃ to obtain a water phase;

2) adding caprylic/capric triglyceride, white oil and stearyl alcohol into an oil phase pretreatment pot, mixing, stirring, and heating to 80-85 deg.C to obtain oil phase;

3) and (4) completely sucking the oil phase into a vacuum emulsifying kettle through filtration, and starting a scraper blade to stir. Then filtering and sucking water phase, after finishing the water phase, sequentially starting rapid stirring and vacuum emulsification, after finishing the emulsification, keeping the temperature and stirring for a few minutes, and then starting cooling water to cool and stir;

4) when the temperature is reduced to about 45 ℃, adding citric acid monohydrate and water, fully and uniformly stirring, and adjusting the pH value of a mixed system to be neutral;

5) sequentially adding nipagin ester, the Fuzhuan tea extract prepared in the embodiment 1 of the invention and citronellol into a mixed system, vacuumizing again, starting a scraper, stirring uniformly, and continuously cooling to below 38 ℃ with cooling water;

6) stopping cooling water, breaking vacuum, carrying out physical and chemical index inspection on the mixed system, filtering and discharging after the mixed system is qualified, and obtaining the anti-aging skin care emulsion.

30 volunteers were selected and randomized for gender. The anti-aging skin care lotion prepared in the example was randomly applied to one of the left cheek or the right cheek and one of the left corner of the eye and the right corner of the eye of each volunteer, respectively, and used as an experimental group; one cheek and canthus of volunteers who did not use the anti-aging skin care emulsion prepared according to the present invention were used as a control group.

The volunteers applied the anti-aging skin care emulsion prepared in the embodiment once in the morning and at night every day, the service cycle was 4 weeks, and no other cosmetics were used in the test cycle. Then, the anti-aging skin care lotion prepared in this example was comparatively analyzed for skin elasticity and wrinkle improvement of volunteers using a skin elasticity tester and a facial wrinkle imager, and the results are shown in table 28:

watch 28

The results show that the anti-aging skin care lotion prepared by the embodiment has very good effects of resisting wrinkles and improving skin elasticity.

Example 16

Examples of cosmetic preparations

The present invention provides several compositions for topical application, particularly for acne.

(1) The formulation of the cleansing cream is shown in table 29:

watch 29

Raw material/brand name	％
		Pure water	QSP 100％
Allan with En (Arlatone)	10-30％
		Cococo glucoside	5-20％
Hydroxypropyl guar gum	1-5％
		Octanoyl glycine	0-2％
Nipagin ester	0-2％
		Citronellol	0-1％
Citric acid	0-1％
		Zinc PCA	0-1％
Fuzhuan tea extract	0.01-5％

(2) The proportions of sebum-regulating anti-acne emulsions are shown in table 30:

watch 30

(3) The formulation of the anti-inflammatory anti-acne emulsion is shown in table 31:

watch 31

(4) The formulation of the restorative anti-acne emulsion is shown in table 32:

watch 32

Raw material/brand name	％
		PEG40 stearate	1％-5％
PEG5 Glycerol stearate	1％-5％
		Earth wax	1％-5％
Glyceryl monostearate	1％-5％
		Sorbitan stearate	0％-2％
Cetyl alcohol	0％-2％
		Malic acid hydrogen ester alcohol	5％-20％
Vitamin E	0％-1％
		Panthenol	0％-5％
Maca peptide extract	1％-5％
		Fuzhuan tea extract	0.01％-5％
Butanediol	1％-5％
		Piroctone olamine	0％-1％
Nipagin ester	0％-1％
		Glycerol	1％-10％
Xanthan gum	0％-1％
		Zinc PCA	0％-2％
Rice starch	1％-5％
		Nylon 6	0％-2％
Polyacrylamide gel	1％-5％
		Vitamin B6	0％-1％
Citronellol	0％-1％
		Pure water	QSP100％

(5) The formulation of the anti-acne cream for regulating the stratum corneum is shown in table 33:

watch 33

(6) The anti-aging and anti-acne emulsion is shown in Table 34:

watch 34

(7) The formulation of the antibacterial anti-acne roll-on stick (stick roll-on) is shown in table 35:

watch 35

Raw material/brand name	％
		Castor oil	QSP 100％
Oleyl alcohol	10％-20％
		Palm oil	10％-20％
Polyglycerol-3-beeswax	10％-20％
		Candelilla wax	10％-20％
Hectorite (hectorite)	10％-20％
		Titanium dioxide	0％-5％
Fuzhuan tea extract	0.01％-5％
		Asiatic fat of snow	0％-5％
Vitamin E	0％-1％

(8) The formula of SPF 50+ anti-acne sunscreen is shown in table 36:

watch 36

(9) The formula of the SPF 50+ anti-acne sunscreen spray is shown in table 37:

watch 37

(10) The compounding ratio of liniment 1 is shown in table 38:

watch 38

Raw material/brand name	％
		Arlatone duo (A La Tong Enduo)	5％-20％
Malic acid	1％-10％
		Sclerotium rolfsii gum	1％-10％
Nipagin ester	0％-1％
		Octanoyl glycine	0％-1％
Soda ash	0％-1％
		Fuzhuan tea extract	0.01％-5％
Acid sodium pyrophosphate	0％-1％
		Citric acid	0％-1％
Pure water	QSP 100％
		Citronellol	0％-1％

(11) The following is the formulation of a specific formula of the liniment, as shown in table 39:

watch 39

Raw material/brand name	％
		Arlatone duo (A La Tong Enduo)	30％
Chinese gum	0.2％
		Glycerol (99%)	2％
Nipagin ester	0.2％
		Sunflower oil	2％
Citronellol	0.2％
		Fuzhuan tea extract	3.4％
Deionized water	62％

The preparation method of the liniment comprises the following steps:

1) heating deionized water to 80 deg.C, dispersing xanthan gum in water phase, and stirring.

2) Slowly stirring and adding Arlatone duo; the temperature was maintained at 75 ℃ with constant stirring, so that the starting material was uniformly dispersed.

3) Adding glycerol, stirring, and cooling.

4) When the temperature is reduced to 35 ℃, the tuckahoe tea extract, the nipagin ester, the sunflower oil and the citronellol are added to complete the preparation of the anti-aging liniment.

40 volunteers were selected and randomized for gender. The anti-aging liniment prepared in the embodiment is respectively and randomly applied to one of the left cheek or the right cheek and one of the left canthus and the right canthus of each volunteer, and is used as an experimental group; one cheek and canthus of the volunteer who did not use the anti-aging liniment prepared by the invention served as a control group.

The anti-aging liniment prepared by the embodiment is applied to volunteers in the morning and at night every day, the use period is 8 weeks, and other cosmetics cannot be used in the test period. Then, the liniment prepared in this example was comparatively analyzed for the skin elasticity and wrinkle improvement of volunteers by using a skin elasticity tester and a facial wrinkle imager, and the results are shown in table 40:

watch 40

Detecting items	Anti-aging liniment
		Wrinkle (wrinkle)	Compared with the control group, the wrinkle depth of the experimental group becomes shallow, and the effect is very obvious
Elasticity of skin	Compared with the control group, the skin elasticity of the experimental group is obviously improved

The result shows that the anti-aging liniment prepared by the embodiment has very good effects of resisting wrinkles and improving the skin elasticity.

Example 17

Preliminary study of Fuzhuan tea extract components: the Jingyang Fuzhuan tea 16KG was pulverized, and 300g of the Fuzhuan tea extract was extracted by the method described in example 1.

Dispersing the Fuzhuan tea extract in 1000mL of distilled water, fully extracting by using petroleum ether, ethyl acetate and n-butyl alcohol in sequence, and carrying out column chromatography on the total extract to obtain a segmented product: petroleum ether extract (2g), ethyl acetate extract (70g), n-butanol extract (81g), and water extract (40g) with small polarity to large polarity.

Example 18

The preparation method of the fu tea organic solvent extract comprises the following steps: grinding Fuzhuan tea into powder, wherein the diameter of tea powder is not more than 0.5cm, adding 85% -95% ethanol into the Fuzhuan tea powder according to the solid-to-liquid ratio of 1:20(m/v), and performing ultrasonic wall breaking until the tea powder and thalli are not visible to naked eyes. Then cold soaking for 18-28 hours, collecting filtrate, repeating for 2-5 times, distilling the filtrate under reduced pressure, concentrating the filtrate, drying at low temperature to obtain crude extract, adsorbing and decolorizing the crude extract by macroporous resin (AB-8 macroporous adsorption resin, Donghong chemical industry), eluting by 85% -95% ethanol, concentrating and evaporating to dryness, and freeze-drying at low temperature to obtain the fu tea organic solvent extract.

Example 19

The preparation method of the Fuzhuan tea water extract comprises the following steps: grinding Fuzhuan tea into powder, wherein the diameter of the tea powder is not more than 0.5cm, adding water into the Fuzhuan tea powder according to the solid-to-liquid ratio of 1:10(m/v), stirring uniformly, and performing ultrasonic wall breaking until the tea powder and the bacteria are not visible to naked eyes. Then standing for 2 hours at room temperature, and boiling for 2 hours in a boiling water bath to obtain a mixed solution; taking out the mixed solution, cooling, centrifuging at an ultra-high speed, and taking a supernatant; filtering and sterilizing the supernatant with 0.22 μm filter membrane to obtain water extract of Fuzhuan tea. Concentrating and evaporating to dryness if necessary, and freeze-drying at low temperature.

Although the present invention has been described with reference to a preferred embodiment, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.

Claims

1. Use of an extract of Fuzhuan tea in the preparation of a skin conditioning product, wherein the skin conditioning product comprises a cosmetic, nutraceutical, dermatological use product against acne;

the Fuzhuan tea extract is supercritical CO₂Extracting the extract;

the supercritical CO₂The preparation method of the extract comprises the following steps: grinding Fuzhuan tea into powder with diameter not more than 0.5cm, completely crushing tea powder and thallus, adding water vapor for pre-soaking until water content is increased to 40%, placing the pre-soaked Fuzhuan tea powder into an extraction tank, and continuously feeding CO into the tank₂Gradually extracting the crude extract with CO of the crude extract₂Sending to a cleaning tank, allowing the Fuzhuan tea extract to enter water phase, and passing through supercritical CO₂Extracting to produce a crude extract, hydrolyzing the starch and fiber with a mixture of cellulase and alpha-amylase, treating the mixture at 65 deg.C for 30 minutes to denature the enzyme, and centrifuging to remove impurities; removing residual proteins by using a 15kDa ultrafiltration column, bleaching the extract with activated carbon and subsequently filtering and returningCollecting, filtering the product with 0.2 μm filter membrane for sterilization, and lyophilizing or packaging to obtain Fuzhuan tea supercritical CO₂And (3) extracting.

2. Use of a fu tea extract according to claim 1 for the preparation of a skin conditioning product, characterized in that the dermatological use product is a dermatological use product against acne.

3. Use of a fu tea extract in the manufacture of a skin conditioning product according to claim 2, wherein the fu tea extract inhibits inflammation by a specific effect against propionibacterium acnes for use as a skin conditioning product.

4. The use of a Fuzhuan extract in the preparation of a skin conditioning product according to claim 1, wherein the skin conditioning product comprises the Fuzhuan extract as the only active ingredient or a pharmaceutical composition comprising the Fuzhuan extract.