CN109010112A - The application of Dermatologic preparation composition containing the ginsenoside Y as principle active component - Google Patents

Abstract

The present invention relates to the applications of the Dermatologic preparation composition containing the ginsenoside Y as principle active component, more specifically, it is related to that there is excellent oxidation resistance containing ginsenoside Y, is capable of providing the effect for improving acne, antibacterial, inhibiting the skin problems such as lipid generation and allergy.

Description

Dermatologic preparation composition containing the ginsenoside Y as principle active component Using

It is on May 2nd, 2014 that the application, which is application No. is the 201480025030.6, applying date, entitled " containing ginseng soap The divisional application of the Chinese patent application of the Dermatologic preparation composition of glycosides Y ".

Technical field

The present invention relates to the Dermatologic preparation compositions for containing ginsenoside Y (ginsenoside Y), more specifically, relating to And containing ginsenoside Y and have excellent oxidation resistance, and can not only provide improve skin moisture-keeping ability effect, Antiphlogistic effects, the effect for improving the skin problems such as acne and allergy, whitening effect, effect, the effect of pore refining for adjusting sebum Fruit improves the effect of the improvement skin integrality such as face by improving blood circulation, but also is capable of providing anti-antidandruff Effect, the effect of the improvement scalp and hair state of educating capillary effect fruit, the effect for preventing white hair etc..

Background technique

First defence film of the skin of people as human body has in the variation of temperature and humidity, ultraviolet light, public hazards substance Etc. the function of intracorporeal organ is protected in the stimulation of external environments, with advancing age, skin is because of a variety of inherent, external factors And undergo variation.That is, inherent is the various hormone secretions reduction for adjusting metabolism, the function and cell activity of immunocyte Decline, so that the biosynthesis of immune protein needed for organism and organism structure protein is reduced, external is because smelly Oxygen layer is destroyed, and the ultraviolet light content that earth's surface is reached in sunlight increases, and as environmental pollution is more serious, free radical and work Property nocuousness oxygen etc. increase, so that skin can cause a variety of variations, that is, not only thickness is thinning, and wrinkle increases, and elasticity reduces, and Skin complexion is dimmed, frequent occurrence skin problem, and spot, freckle and senile plaque can also increase, and complexion is deteriorated and the colour of skin is dimmed.

This skin is because of inherent and external factor skin condition variation in order to prevent, and maintains the skin shape of health State, the effort done always are will to be derived from being added to of physiological activator of conventionally known various animals, plant, microorganism etc. It uses in cosmetic, so as to improve skin condition, has especially done much in cosmetics using ginseng (korean ginseng, Panax Ginseng C.A.Meyer) ingredient research and development.Ginseng is certified the function of skin already, to be widely used In cosmetic composition, the root of ginseng or extract, the ginsenoside (ginsenoside), panaxcoside aglucon of leaf (aglycon) it is often used with panaxan's body etc..But contain the makeup of ginseng extract, essence, polysaccharide body etc. at these In product, contained by active material be denier, to have the drawback that, compared with other raw materials, effect is not very Obviously.

Summary of the invention

Technical task

For this purpose, the inventors discovered that the ginsenoside Y contained by ginseng can not only provide whitening, moisturizing improves effect Fruit, but also it is capable of providing the effect for improving the improvement skin condition such as effect, the effect for adjusting sebum, pore refining of complexion, In addition it is possible to the effect of the improvement scalp and hair state that anti-antidandruff are provided, hair is educated and prevents white hair etc., so as to complete The present invention.

Therefore, the purpose of the present invention is to provide a kind of skins that can improve skin integrality containing ginsenoside Y Skin preparation composition for external use.

Project solution

In order to achieve the above object, the present invention provides a kind of external preparation for skin containing the ginsenoside Y as effective component Agent composition.

In addition, the present invention provide it is a kind of containing as effective component ginsenoside Y be used for whitening skin preparations for extenal use group Close object.

In addition, the present invention provide it is a kind of containing as effective component ginsenoside Y be used for moisturizing skin preparations for extenal use group Close object.

It is used to improve the external preparation for skin of acne in addition, the present invention provides a kind of ginsenoside Y containing as effective component Agent composition.

It is used to improve the external preparation for skin of allergy in addition, the present invention provides a kind of ginsenoside Y containing as effective component Agent composition.

It is used to improve the skin of complexion and the colour of skin in addition, the present invention provides a kind of ginsenoside Y containing as effective component Skin preparation composition for external use.

In addition, the present invention provide it is a kind of containing as effective component ginsenoside Y be used for pore refining external preparation for skin Agent composition.

It is used to adjust the external preparation for skin of sebum in addition, the present invention provides a kind of ginsenoside Y containing as effective component Agent composition.

In addition, the present invention provide it is a kind of containing as effective component ginsenoside Y be used for anti-antidandruff external preparation for skin Agent composition.

It is used to educate the skin preparations for extenal use group of hair in addition, the present invention provides a kind of ginsenoside Y containing as effective component Close object.

It is used to prevent the external preparation for skin of white hair in addition, the present invention provides a kind of ginsenoside Y containing as effective component Agent composition.

In addition, the present invention provides a kind of Dermatologic preparation composition for using ginsenoside Y for natural antiseptic agent.

Beneficial effect

Composition of the invention has excellent oxidation resistance due to containing ginsenoside Y, and can not only provide Improve effect, antiphlogistic effects, the effect for improving the skin problems such as acne and allergy, whitening effect, adjusting of skin moisture-keeping ability The effect of sebum, the effect of pore refining, improve because improving blood circulation face etc. improvement skin integrality effect, But also what is be capable of providing the effect of anti-antidandruff, educate capillary effect fruit and prevent the effect of white hair etc. improves scalp and hair state Effect.

Specific embodiment

Dermatologic preparation composition according to the present invention contains the ginsenoside Y as effective component, particularly the people Ginseng saponin(e Y can be extracted from ginseng.

There is the structure of following chemical formula 1 for ginsenoside Y of the invention

Chemical formula 1

Ginseng belongs to the Panax of Araliaceae (Araliace), is the dicotyledonous herbaceous plant of Su Xinggen.South Korea, China, day There are production in multiple countries such as this and the U.S., and Main Cultivation area is: it is east longitude 85-140 degree, north latitude 22-49 degree in East Asia, North America is west longitude 70-97 degree, north latitude 34-47 degree.The ginseng so produced all over the world difference on ingredient, display Most importantly it is known as the ginseng soap of ginsenoside (ginsenoside) when the various physiology of ginseng and effect pharmacologically Plain (saponin), the content that South Korea produces ginseng saponin in ginseng are most.Up to the present, the ginsenoside isolated is about 34 Kind, according to be incorporated on aglycone sugar type, in conjunction with sugar quantity or binding site, pharmacological effect is not Together.According to the feature in structure, ginsenoside is divided into protopanoxadiol (Protopanaxadiol) type, protopanaxatriol (Protopanaxatriol) type and oleanane (oleanane) type.In addition, also containing as the intrinsic fragrance component of ginseng Panacene (panacen), polyacetylene compound, polyphenolic substance, flavonoids and vitamin etc..

The Chinese medicine effect of ginseng is just recognized from ancient times, for treating a variety of diseases.It is known in the Chinese medicine effect of ginseng Be enhancing physical strength, improve metabolism, relieve pressure, treatment diabetes, improve respiratory disease, improving digestive organs and anti- Cancer effect, disclose in a recent study ginseng with anti-complement activity, anti-ulcer effect, enhancing be immune, antitumaous effect and Anti- blood glucose decline effect etc..Studying effect relevant to skin in delivering includes antiinflammation (Korean J.Dermatol.18 (1): 39-42 (1980), Korean J.Dermatol.14 (4): 335-339 (1976)), hyperkeratosis (Korean is prevented J.Dermatol.28 (4): 434-440 (1990)) acne (Korean J.Dermatol.30 (1): 27-33, is prevented and treated (1992), (4) Korean J.Dermatol.28: 434-440 (1990)), antioxidant effect (Proceedings of the 2nd international ginseng symposium 13-17 (1978)), enhancing elasticity and hydrability (Fitoterapia 57 (1): 15-28 (1986), Fitoterapia 57 (4) 217-222 (1986)), treat wound (Brit.J.Pharmacol.125:255-262 (1998), Arch.pHarm.res.25 (1): 71-76 (2002)), inhibit glue Former protein breakdown (Mol.Cells 9 (5): 476-483 (1999)), whitening effect (Journal of the society of (2001) cosmetic scientists of korea 27 (2): 45-56) etc..

Ginsenoside Y of the invention can be extracted from plants, and can also synthesize and make by methods known in the art With commercial product can also be used.Additionally, it is contemplated that when the technical level of this field, those skilled in the art it is clear that, pass through To the ginsenoside Y carry out the substituent group usually implemented in this field substitution and displacement reaction and obtain inductor in, Display improves skin, the effect of hair and the inductor of anti-corrosion effect and is also included in the scope of the present invention.

Ginsenoside Y used in the present invention can especially extract from ginseng extract.Wherein, the ginseng kind used Class is not particularly limited, and water ginseng, red ginseng, white ginseng, Tai Ji ginseng, tail ginseng etc. can be used.In addition, the ginseng extract includes The leachate for leaching, producing and obtaining from ginseng, leachate is partly or entirely concentrated again and the concentrate that obtains, Or the concentrate is dried again and being contained in for manufacturing be stagnant, decoction, tincture, sends out in fluid essence and ginseng The chemical substance of main effect is waved, certainly, may also comprise plant itself, the ginsengs such as stem, root, leaf, flower, fruit can be used The extract of all sites is not limited to the extract of some privileged site.In addition, extracting ginsenoside from ginseng extract The method of Y can use well known method.

Specifically, by method commonly used in the art, ginseng is produced from ginseng using water or organic solvent and is extracted After object, and from wherein isolating the ginsenoside Y.Organic solvent used in the present invention can be from by ethyl alcohol, methanol, fourth It is selected in the group that the mixed solution of alcohol, ether, ethyl acetate, chloroform and these organic solvents and water is constituted, it is preferred to use 80% Ethyl alcohol.At this point, Extracting temperature is preferably 10~80 DEG C, 3~24 hours can be extracted.When beyond the Extracting temperature and mentioning When taking the time, extraction efficiency may be reduced or be changed at branch.

The content of ginsenoside Y contained by composition according to the present invention accounts for 0.001 in the total weight of composition~ 50 weight %, preferably 0.01~30 weight %, more preferably 0.1~10 weight %.When the ginsenoside Y content less than , can be faint according to the effect of the ingredient, effect when 0.001 weight %, when more than 50 weight %, there are cutaneous safety or Problem in dosage form.

Ginsenoside Y is the ingredient with excellent oxidation resistance, therefore, the group of the invention containing ginsenoside Y It closes object and excellent oxidation resistance is provided to skin.

Composition of the invention can be used as the composition of whitening, because it is able to suppress tyrosinase activity and inhibits black The generation of element, therefore it is capable of providing excellent whitening effect.

Composition of the invention can be used as the Dermatologic preparation composition of moisturizing, can strengthen skin barrier function, And it is capable of the differentiation of induced skin keratinocyte.Therefore, can effectively with for prevent or improve because of epidermal differentiation not The Dermatologic preparation composition of the xerodermia, allergic dermatitis, contact atopic dermatitis or the psoriasis that generate completely etc..

Composition of the invention can be used as the Dermatologic preparation composition of acne improvement, provide antibacterial effect, especially It is excellent to the antibacterial effect of acne pathogenic bacteria, in addition, also providing antiphlogistic effects.

Composition of the invention can be used as improving the Dermatologic preparation composition of complexion and face, when being used on skin Capillary can be expanded and promoted blood circulation, so that nutritional ingredient is swimmingly provided to skin and inhibits skin aging, because This, improves the significant effect of complexion and the colour of skin.

The skin preparations for extenal use combination that composition of the invention can be used as pore refining, adjust sebum and improve skin problem Object when being used on skin, inhibits Excess sebum production, and by promoting the removal of active oxygen and the synthesis of collagen, from And pore refining, since the expression of pro-inflammatory cytokine is reduced, to inhibit the excellent effect of skin problem.In addition, because excellent resists Oxidability can prevent the generation of skin irritatin.

Group of the invention is combined into the Dermatologic preparation composition that can be used as anti-antidandruff, can be effectively discharged out accumulation Toxin on hair and scalp and purify scalp, the proliferation and growth of dandruff bacterium are able to suppress, so that it is anti-to prevent scalp inflammation It answers, in addition, being capable of providing stable scalp and reinforcing since the antioxidation of generation and the effect of inhibitory activity oxygen is significant Scalp, and the effect for strengthening original defence capability is provided.

Composition of the invention can be used as educating the Dermatologic preparation composition of hair, by promoting from hair week dormant period Phase is transferred to growth period hair cycles, to promote the growth of hair, and provides the effect prevented hair loss.

Composition of the invention can be used as preventing the Dermatologic preparation composition of white hair, by make melanocyte to The expression of MITF increases significantly, to inhibit white hair and induce long dark hair.

In addition, being capable of providing the effect of natural antiseptic agent for the ginsenoside Y in Dermatologic preparation composition of the invention Fruit.

The Dermatologic preparation composition of aforementioned present invention as change the outfit product composition when can dosage form, may include makeup Allow in conduct and learning or Dermatology using medium or base and dosage form.It can be using all doses be suitble to locally Type, for example, can be provided as solution, gel, solid, the anhydrous product of stirring, in the liquid phase dispersed oil phase and obtain lotion, The form of the vesica dispersing agent of suspension, microemulsion, microcapsules, microparticle ball or ionic (liposome) and nonionic, Or it also can be provided as the forms such as face cream, toner, skin care milk, foundation cream, ointment, spraying or concealer.Further, it is also possible to make With the form of the aerosol composition of the propellant for foam (foam) shape or further containing compression.These compositions can It is manufactured by usual way in this field.

It particularly, can agent when Dermatologic preparation composition of the invention is used for anti-antidandruff, educates hair or prevent white hair Type chemical conversion is used for the composition of scalp and hair, and dosage form is not by particular determination, for example, hair restorer, hair battalion can be turned to dosage form Support toner, scalp nursing solution, hair shield liquid reason, shampoo, hair conditioner, hair cream or scalp and hair-care dual-purpose conditioning liquid etc..

In addition, composition according to the present invention can contain fatty material, organic solvent, lytic agent, concentrating agents, gelation Agent, softening agent, antioxidant, suspending agent, stabilizer, foaming agent (foaming agent), aromatic, surfactant, water, Ionic or non-ionic softening agents, metal ion block agent, chelating agent, antistaling agent, vitamin, blocking agent, moisten filler Agent, essential oil, dyestuff, pigment, hydrophily or lipophilic activating agent, lipid vesicle or with it is usually used arbitrary in cosmetics Usually used adjuvant in the identical cosmeceutical of other compositions or Dermatology field.The amount of the adjuvant being added is The amount generally used in cosmeceutical or Dermatology field.

In addition, composition of the invention in order to increase skin improvement effects, can contain the substance for promoting skin to absorb.

Hereinafter, more particularly illustrating structure and effect of the invention by experimental example and dosage form example.But these are tested Example and dosage form example are only used for helping to understand that the present invention, scope of the invention and range are not limited by following examples as illustration.

Reference example 1

The ginsenoside Y of effect for testing composition of the invention is to buy and use from AMBO research institute.

Test example 1

The effect that inhibitory activity oxygen (ROS:reactive oxygen species) generates

The Mei Gekong into 24 orifice plates is added in the keratinocyte (keratinocyte) of removing from the epidermal tissue of people 5 × 10 are added in plate⁴It is a, carry out 24 hours of attachment.After 16 hours, ginsenoside Y is processed into 1%.At this point, in order to do Compare, without processing ginsenoside Y in control group.After two hours, culture solution is removed, adds 100 μ l in each hole later Phosphoric acid buffer saline solution (PBS).Using Ultraviolet B (UVB) lamp (Model:K5T8, UV B 15W, Sankyo Dennki company, Japan), 30mJ/cm is irradiated to the keratinocyte²Ultraviolet light after, isolate PBS, and 200 μ l are added in each hole Keratinocyte culture solution.Here, ginsenoside Y is handled again, according to certain time interval measurement because ultraviolet light pierces The content of sharp and increased active oxygen.When measuring the content of ROS with reference to the method for Tan (Tan et al., 1998, J.Cell Biol.Vol.141, pp1423-1432), that is, by measure the dichlorodiethyl acid esters aoxidized by ROS (DCFDA, Dichlorofluorescin diacetate) fluorescence spectrum measure the content of ROS, calculate for only handling excipient Control group ROS ratio, result is see the following table 1.

Table 1

It is found that ginsenoside Y according to the present invention can effectively inhibit the generation of ROS, ROS from the result of upper table 1 It is the well known substance for causing Skin Cell to damage, it can be more when the amount of the post-stimulatory ROS of ultraviolet light is not than irradiating ultraviolet light Inhibit the generation of ROS, to know that antioxidant effect is very prominent.Therefore confirm, ginsenoside Y according to the present invention can Inhibit oxidation, prevent aging, broaden so as to prevent pore, and the generation by preventing skin irritatin improves skin and asks Topic.

Test example 2

Tyrosinase inhibitory effect

Tyrosinase is extracted from mushroom class (Mushroom), and the product of Sigma (Sigma) company is used.It is first First, the tyrosine as matrix is dissolved in distilled water, to be configured to the solution of 0.3mg/ml, into each developmental tube The above-mentioned solution of 1.0ml is added, adds potassium-phosphate buffer (0.1 molar concentration, pH 6.8) of 1.0ml thereto later With the distilled water of 0.7ml.

Ginsenoside Y of the invention is mixed in ethanol solution with debita spissitudo, and it is ready each to add 0.2ml Sample liquid is reacted 10 minutes in 37 DEG C of thermostat later.At this point, control group is prepared to only add each sample liquid of substitution The solvent of 0.2ml, positive controls use ascorbic acid.2500 units/ml junket ammonia of 0.1ml is added in the reaction solution Sour enzyme solutions react 10 minutes again in 37 DEG C of thermostat.Developmental tube equipped with the reaction solution is placed in ice water and is carried out It is quickly cooled down to terminate reaction, and the absorbance in 475nm wavelength is measured by photoelectricity spectrum analysis instrument, result is see following table 2.Each tyrosinase inhibitory action is calculated by following mathematical expression 1.

Mathematical expression 1

Table 2

Substances	Tyrosinase inhibition rate (%)
		Control group (is not added with)	0
Ascorbic acid	52
		Ginsenoside Y	59

In the result of upper table 2, the tyrosinase inhibition rate ratio of ginsenoside Y according to the present invention is known as junket ammonia The ascorbic acid of sour enzyme inhibitor is much higher, to know that whitening effect is very excellent.

Test example 3

Inhibit the effect of melanin production by using B16/F10 melanoma cells

Substances are added to using the sample of ginsenoside Y and kojic acid respectively containing 0.1 weight % with a certain concentration In the culture solution of B16/F10 melanoma cells (Korea Cell strain bank), after culture 3 day, culture solution is removed, is washed with PBS, Cell is dissolved with the NaOH of 1N, absorbance is measured in 405nm.As a control group by the substances for being not added with cell, pass through It is compared with the melanin content in control group, to measure the degree of the inhibition melanin production of each substances.Root Melanin is calculated according to mathematical expression 2 and generates inhibiting rate, and result is see the following table 3.

Mathematical expression 2

Table 3

Substances	Melanin generating suppression (%)
		Control group (is not added with)	0
Kojic acid	53
		Ginsenoside Y	65

In the result of upper table 3, the melanin generating suppression ratio of ginsenoside Y according to the present invention is known as black The kojic acid of pigment formation inhibitor is higher, to know that whitening effect is very excellent.

Dosage form example 1 and compare dosage form example 1

According to the composition of the following table 4, nourishing cream (unit: weight %) is manufactured using usual method.

Table 4

Test example 4

The effect of measurement enhancing skin moisture-keeping power

In order to measure ginsenoside Y to the effect of enhancing skin moisture-keeping power, using above-mentioned dosage form example 1 and compare dosage form example 1, And make following evaluation.

20 people of adult men and women that would be classified as 40-50 years old of drying property skin, is divided into two groups, every group of 10 people, and needle respectively Nourishing cream is being smeared to dosage form example 1 and two groups for comparing dosage form example 1 on the face, is being smeared daily twice, totally 4 weeks.Start in smearing Before, smear after by 1 week, 2 weeks, 4 weeks and after termination smearing after 2 weeks (in total by 6 weeks), in constant temperature, constant humidity condition Under (24 DEG C, relative humidity 40%), using skin moisture content measuring instrument (Corneometer CM825, C+K Electronic Co., Germany) measurement skin moisture content.It is in table 5 the result is that just to start to try the result is shown in the following table 5 The measured value of the skin moisture content measured before testing is as benchmark, with the percentage of the incrementss by measured value after a certain period of time Rate is indicated.

Table 5

The result for seeing the above table 5 is confirmed, when smearing compares dosage form example 1, shows that about 30% moisture increases when smearing 4 weeks Add rate, but after terminating smearing, skin moisture content is sharply reduced, in contrast to this, when dosage form of the smearing containing ginsenoside Y It when example 1, terminates after smearing, major part still shows 30% or more moisture of skin increment rate.It follows that containing ginseng The skin moisturization of the composition of the invention of saponin(e Y is excellent.

Test example 5

Measurement promotes the effect of Keratinocyte differentiation

In order to understand the effect for promoting Keratinocyte differentiation of ginsenoside Y, angle is measured using absorbance below Matter forms the content of the hornification coating (CE, Cornified Envelop) generated when cell differentiation.

Firstly, the keratinocyte for the people that will be isolated from neonatal epidermis and be once cultivated is added to culture bottle In, and it is attached to keratinocyte in bottom of bottle, ginsenoside Y is processed into after the concentration of 5ppm in culture solution, is trained Support 5 days until cells grown to flat area 70~80% degree.At this point, by low calcium (0.03mM) processing group and high calcium (1.2mM) processing group is used separately as negative control group and positive controls.Later, the above-mentioned cell turned out is extracted, using phosphorus Hydrochlorate buffer saline (PBS, PhopHate buffered saline) washed, and that adds 1ml later contains 2% Lauryl sodium sulfate (SDS, sodium dodecyl sulfate) and 20mM concentration dithiothreitol (DTT) (DTT, Dithiothreitol the Tris- hydrochloride buffer (Tris-HCl, pH 7.4) of 10mM concentration), and carry out ultrasonic wave (sonication), (boiling), centrifugation are boiled, sediment is precipitated in the PBS of 1ml again, to measure Absorbance in 340nm.Individually with aforesaid operations, the part solution after carrying out the ultrasonic wave is taken, protein content is measured, And benchmark when being evaluated as cell differentiation.The result is shown in the following table 6.

Table 6

Substances	Differentiation capability (%) in keratinocyte
		Low calcium (0.03mM) solution (negative control group)	100
High calcium (1.2mM) solution (positive controls)	210
		Ginsenoside Y	290

Shown in table 6 as above, it is capable of determining that when handling ginsenoside Y, promotes the effect of Keratinocyte differentiation excellent It is different.

Test example 6

Measurement restores the effect of skin barrier

In order to measure the effect that ginsenoside Y restores the skin barrier function of damaging by skin injury, carry out as follows Experiment.The upper arm of adult 10 people of men and women is given skin barrier and is damaged using adhesive tape removing (Tape Stripping) method, And it smears respectively and the dosage form example 2 manufactured and two groups for comparing dosage form example 2 is formed by table 7, and in 7 days, use The recovery extent of a skin water loss amount (TWEL) is compared in Vapometer (Delfin, Finland) measurement daily.Wherein, than Being used as negative control group compared with dosage form example 2 is excipient (vehicle).Experimental result is shown in the following table 8.In the result of table 8, damaged with barrier Wound before and barrier damage after processing before difference as 100% benchmark and compare.

Table 7

Gradation composition	Dosage form example 2	Compare dosage form example 2
			Pure water	69	70
Propylene glycol	30	30
			Ginsenoside Y	1	-

Table 8

It is capable of determining that from upper table 8, when processing does not contain the comparison dosage form example 2 of ginsenoside Y, with the stream of time It dying, skin water loss amount is increasing, in contrast to this, and when handling the dosage form example 2 containing ginsenoside Y, moisture of skin Loss amount can fast quick-recovery it is normal, barrier damage can restore.

Test example 7

Improve the effect of complexion

In order to evaluate the effect that cosmetic composition according to the present invention promotes skin blood circulation, using laser-Doppler Blood flowmeter (LDPI, Laser Doppler Perfusion Imager) measures the blood circulation degree in skin.LDPI is public The sanguimotor equipment in measurement skin known, and as currently used device, it is that one kind can not only determine skin The flow velocity and flow of blood in skin capillary, but also the very clever of flowing in parteriole and veinlet can be determined Quick equipment.

It in thermostatic constant wet chamber, is adapted to 30 minutes after being washed one's face with perfumed soap, initial value is measured using LDPI.Firstly, using LDPI Measure the initial blood flow of the forehead lower part of 30 ice-cold women of usually trick.Later, subject is allowed to use above-mentioned dose respectively Type example 1 and two groups for comparing dosage form example 1 will measure blood flow using 1 week later, the ratio of the blood flow and the initial value Relatively result (skin blood flow variation) see the table below 9.

Table 9

Use LDPI result-skin blood flow before and after cosmetics

Substances	Use the skin blood flow change rate (%) after 1 week
		Dosage form example 1	11
Compare dosage form example 1	5

It is capable of determining that in the result of upper table 9, cosmetic composition according to the present invention and the ratio for being free of ginsenoside Y It is compared compared with dosage form example 1, skin blood flow dramatically increases, therefore this can improve complexion by promoting blood circulation.This final table The bright cosmetic composition containing ginsenoside Y of the invention can effectively transmit the nutriment in skin, and can Inhibit and postpones skin aging.

Test example 8

Improve the effect of the colour of skin

In order to understand the effect of above-mentioned dosage form example 1 and the improvement colour of skin for comparing dosage form example 1,30 subjects is allowed to make respectively After (smearing at night, 1 times/day, totally 1 week), improved using Facial Stage DM-3 (Moritex, Japan) evaluation colour of skin Degree.Colour of skin improvement rate uses the brightness of skin and the brightness of the measured value of color and skin and color change value to be judged, The result is shown in the following table 10.Brightness and color change value show that more greatly the colour of skin is improved.

Table 10

It is determined in the result of table 10, the comparison dosage form example 1 without ginsenoside Y of the invention is not shown obviously Colour of skin improvement, in contrast to this, containing in the dosage form example 1 as the ginsenoside Y of effective component, than using after use The preceding colour of skin improves very much.

Test example 9

The effect of pore refining

1. the effect of the pore refining by promoting collagen synthesis

By comparing ginsenoside Y according to the present invention and TGF-β, to measure the effect for promoting collagen synthesis. Firstly, sowing (seeding) 10 in each hole of 24 holes (well)⁵A fibroblast (fibroblast), and cultivate extremely The degree that cells grown is 90%.After plasma-free DMEM medium culture 24 hours, it will be dissolved in serum free medium Ginsenoside Y of the invention and TGF-β are processed into 10 μ g/ml respectively, and in CO₂24 hours are cultivated in incubator.Take out this The upper liquid of a little solution observes precollagen using I procollagen type ELISA kit (procollagen type (I)) (procollagen) whether increase and decrease.The result is shown in table 11, untreated fish group is set as 100, the synthesis performance of collagen with 100 untreated fish groups compare.

Table 11

Substances	Collagen synthesis performance (%)
		Untreated fish group	100
TGF-β	183.5
		Ginsenoside Y	189.9

It is capable of determining that in the result of upper table 11, ginsenoside Y according to the present invention is more aobvious than the TGF-β of positive controls Higher and excellent collagen synthesis performance is shown.Therefore, it is capable of determining that ginsenoside Y according to the present invention can pass through Increase the collagen production quantity around pore and shrinks the pore to become larger.

2. the effect of pore refining

In order to understand the effect of dosage form example 1 with the pore refining for comparing dosage form example 1, following evaluation has been carried out.Selected pore Big 20 people of subject men and women, is divided into two groups, every group of 10 people, and is smearing dosage form example 1 daily according to each group on the face and comparing The nourishing cream of dosage form example 1 is smeared 4 weeks altogether.It takes a picture before experiment and respectively after 4 weeks, by the evaluation with the naked eye of expert, to judge to receive The effect of contracting pore.The result is shown in the following table 12, (opinion rating: 0- is absolutely not shunk；5- shrinks very much).

Table 12

Substances	Opinion rating
		Dosage form example 1	4
Compare dosage form example 1	0

It is found that comparing does not have the effect of pore refining in dosage form example 1 from upper table 12, however then shown in dosage form example 1 The pore contractive effect for the degree that can with the naked eye determine, therefore, ginsenoside Y according to the present invention reduce the effect of pore size Fruit is excellent.

Test example 10

Inhibit the effect of sebum secretion

1. by inhibiting 5α-reductase activity to inhibit the effect of skin excessive secretion

Inhibit the active effect of 5α-reductase to determine, measures in HEK293-5 α R2 cell, [¹⁴C] testosterone turn Become [¹⁴C] protona ratio.P3 × FLAG-CMV-5 α R2 is transfected in HEK293 cell, and is added into 24 orifice plates It is cultivated, each hole addition 2.5 × 10⁵A cell (Park et al., 2003, JDS.Vol.31, pp.191-98).Second It, is replaced with the new culture medium added with enzyme matrix and inhibitor.The matrix of culture medium use 0.05 μ Ci [¹⁴C] testosterone (Amersham Pharmacia biotech,UK)。

Inhibit the active degree of 5α-reductase to determine, ginsenoside Y is added, and in 37 DEG C, 5%CO₂Incubator 2 hours of middle culture.At this point, the culture solution for being not added with ginsenoside Y is used as negative control group, Finasteride will be added with (finasteride) culture solution is as positive controls.Later, culture solution is extracted, class is extracted using the ethyl acetate of 800 μ l Sterol separates the organic solvent layer on top and drying later, dissolves remaining residual using the ethyl acetate of 50 μ l again later Object, with acetic acid on silica plastic plate silica gel 60F254 (Silica plastic sheet kieselgel 60F254) Ethyl ester-nucleic acid (1:1) is unfolded as solvent.

Plastic sample is dried in air, in order to measure isotopic content, using Bath system, wherein will do Dry plastic sample and X-ray film is added together in Bath box, and measurement remains in testosterone and double hydrogen on film after a week The isotopic content of testosterone calculates separately out conversion ratio and inhibiting rate, result according to following mathematical expressions 3 and mathematical expression 4 later See the following table 13.

Mathematical expression 3

Mathematical expression 4

Table 13

Substances	Conversion ratio (%)	Inhibiting rate (%)
			Negative control group	48.0	-
Positive controls	27.6	42.5
			Ginsenoside Y	13.9	62.8

It is able to confirm that out in the result of above-mentioned table 13,5α-reductase makes testosterone be converted into protona and and cell Receptor protein in matter combines, into nucleus in and activate sebocyte cell and promote to break up, so that 5α-reductase has Play the role of playing excessive secretion sebum in sebaceous glands, and ginsenoside Y can effectively inhibit the activity of 5α-reductase, So as to block testosterone to be converted into protona, show than the well known active Finasteride for inhibiting 5α-reductase With superior inhibitory effect.Therefore, ginsenoside Y is able to suppress sebum by effectively inhibiting the activity of 5α-reductase Secretion is excessive.

2. inhibiting the effect of sebum secretion

In order to understand above-mentioned dosage form example 1 and compare the effect for inhibiting sebum secretion of dosage form example 1, following evaluation has been carried out. Selected 30 people of subject men and women thought more than sebum secretion, and smear dosage form example 1 daily on specified position and compare dosage form The nourishing cream of example 1 is smeared 4 weeks.Using sebum flowmeter (Sebumeter SM810, C+K Electronic Co., Germany) The average sebum slip (%) after 2 weeks and after 4 weeks is measured, respectively to judge the effect of sebum reduction, result It see the table below 14.

Table 14

It is found that the dosage form example 1 containing the ginsenoside Y according to the present invention as effective component from the result of upper table 14 Sebum excessive secretion can more effectively be inhibited than the comparison dosage form example 1 without ginsenoside Y.

Dosage form example 3 and compare dosage form example 3-4

The ingredient according to shown in the following table 15 and content (weight %) have prepared dosage form example 3, have compared dosage form example 3 and compare Dosage form example 4.If illustrating, it is combined with ginsenoside Y in dosage form example 3, compares dosage form example 3 absolutely not comprising improving acne The effective component of skin compares dosage form example 4 and contains the acne therapeutic agent that is commonly used for as the standard substance that judges anti-microbial property Erythromycin (erythromycin).

Dosage form example 3, the manufacturing method for comparing dosage form example 3 and comparing dosage form example 4 are as follows.It is completely dissolved in the A of the following table 15 Ingredient is completely dissolved after the ingredient in B in additional dissolving tank, and B is added in A and carries out mixing solubilization solution.At this point, Ingredient in C is added according to documented configuration proportion in table 15, is filtered after being uniformly mixed, to make This composition.

Table 15

Test example 11

For the antibacterial ability test of acne bacterium

It uses with dosage form example 3, compare dosage form example 3 and compare each cosmetic composition prepared that forms of dosage form example 4, it is right Propionibacterium acnes (ATCC 6919: culture medium-BHI fluid nutrient medium (broth)) as acne pathogenic bacteria have carried out antibacterial Ability test.

It is as follows for the test method of the antibacterial ability of acne bacterium.

(1) prepare test bacteria liquid

Propionibacterium acnes use the culture solution for being inoculated with and carrying out Anaerobic culturel in BH1 fluid nutrient medium.

(2) prepare dilute solution

The addition 0.15ml in the BH1 fluid nutrient medium (pH 6.8) or LB liquid medium (pH 4.5) of 15ml Uniformly mixed solution is used as dilute solution by above-mentioned test bacteria liquid.

(3) prepare sample

Sample will be used as by dosage form example 3, the original solution for comparing dosage form example 3 and comparing the cosmetic composition that dosage form example 4 manufactures Product.

(4) antibacterial ability is tested

1) sample is added with meeting initial concentration in the first row of 96 hole cell culture tubes (96well plate), and Dilute solution is added until total amount is 200 μ l.

2) it uniformly takes 100 μ l to be added in the second row and uniformly mix after the mixed liquor of mixing the first row, takes again later 100 μ l are added in the third line, in the above way progress doubling dilution (double dilution).

3) stationary culture 24 hours and 48 hours at 32 DEG C, judge whether bacterium is proliferated into suspended degree later, and will Bacterium is determined as MIC (minimum inhibitory concentration, Minimum Inhibitory Concentration) value without the Cmin of proliferation. When being difficult to judge whether bacterium is proliferated if mixed liquor is opaque, confirmed using micro- sem observation.

For acne bacterium antibacterial ability test result see the following table 16.MIC be converted into be contained in dosage form it is effective at The concentration divided is marked.

Table 16

Project	pH	Propionibacterium acnes
			Dosage form example 3	5.7	>50ppm
Compare dosage form example 3	5.7	Maximum concentration (does not have antibacterial ability)
			Compare dosage form example 4	5.7	>100ppm

In MIC, the smaller more effective substance of antibacterial ability being shown to be for acne bacterium of concentration in dosage form example 3 and is used The comparison dosage form example 4 for being known as the erythromycin of acne therapeutic agent is compared, and ppm concentration may be significantly smaller, and therefore, be capable of determining that Composition containing ginsenoside Y has superior antibacterial ability to test organisms.

Test example 12

Inhibit the test of lipid synthesis (Lipogenesis)

By as the 3T3-L1 cell of the fibroblast (fibroblast cell line) of mouse living with 1 × 10⁵'s Amount is attached on 6 well culture plates (culture plate), is added on 6 well culture plate and is contained fetal calf serum (FBS, fetal Bovine serum) DMEM (Dulbecco's modified eagle's medium, GIBCO BRL, Life Science are public Department) culture medium.After 2 days, changes new DMEM (FBS containing 10%) culture medium into, be further cultured for 2 days.Later, by containing 1 μ The insulin (insulin) of g/mL, the DMEM of IBMX and 0.25 μM of the dexamethasone (dexamethasone) of 0.5mM (contain 10% FBS), induce the cell of above-mentioned culture to be broken up again, the ginsenoside Y and caffeine of 50 μM of processing, by 2 days Afterwards, it changes the DMEM containing insulin into again, cultivates 5 days.After 5 days, normal incubation medium (DMEM, containing 10% is changed into again FBS), observation culture to above-mentioned cell becomes fat cell in shape.

Inhibit the effect that fat is accumulated in fat cell to evaluate ginsenoside Y, using above-mentioned differentiated 3T3-L1 Fat cell has carried out soudan III dyeing (S4136, sigma-aldrich) to it.At normal temperature, by fat cell poly Formaldehyde (pH 7.2) is fixed in phosphate buffer, is washed later with PBS (phosphate buffered saline), it It is dyed with soudan III, is taken a picture later afterwards, and be compared by naked eyes.Control group, which only used, is not added with substances or ratio 50 μM of caffeine has been handled as other comparative groups compared with the culture medium of substance.Inhibit the degree of lipopexia by that will dye Degree is divided into +++, ++ ,+,-impart grade.Wherein, more past +++ it walks, indicates that dye levels are bigger.The result is shown in the following table 17.

Table 17

Sample	Inhibiting rate (%)
		Control group	+++
Comparative group	+
		Ginsenoside Y	-

It is found that the ginsenoside Y used in the present invention not only makes the fat accumulated in fat cell shown in the upper table 17 Content is few, and the caffeine than being known as lipid synthesis inhibiting substances has the superior effect for inhibiting lipid synthesis Fruit.Therefore, by inhibiting lipid synthesis, sebum can be reduced, so as to inhibit the generation of acne.

Test example 13

Improve acne, reduce sebum secretion and have non-stimulated test

30 people for having acne are divided into 3 groups, every group of 10 people use corresponding subject respectively above-mentioned by agent Type example 3, the cosmetic composition for comparing dosage form example 3 and comparing the manufacture of dosage form example 4, use 1 month.The scale setting that acne improves It is 1 point to 5 points, 1 point of expression " not having ", 3 points of expressions " general ", 5 points of expressions " very good ".Experimental result is shown in the following table 18, wherein It is marked with the average mark of 10 people.

Acne disappears period to judge the date to disappear as benchmark, and acne recurrence uses " whether there is or not " with the knot after 1 month Fruit is as benchmark.Sebum secretion reduction is set as 1 point to 5 points, 1 point of expression " not having ", 3 points of expressions " general ", and 5 points of expressions are " non- It is often good ".Experimental result is see the following table 22, wherein being marked with the average mark of 10 people.Whether there is or not skin irritatin pass through it is (irritating anti- The number answered)/(overall test person number) observe.

Table 18

It is found that the dosage form example 3 compared with of dosage form example 3 is compared shown in the upper table 18, acne does not recur, on the whole to improvement Acne has the effect of excellent.In addition, showing that acne improves effect in the comparison dosage form example 4 containing antibacterial ability standard substance Fruit, but stimulation is big when using, therefore is not suitable for being used for a long time, and composition according to the present invention is shown without stimulation and fits It closes and is used for a long time.

Test example 14

The effect for inhibiting IL-8 to generate

Before experiment one day, by Keratoderma epithelial cell (NHEK, Normal human skin keratinocyte；Purchase Buy place-Lonza) on 96 orifice plates plant division at 5 × 10⁴Cell number/hole, later in 37 DEG C, 5%CO₂Incubator in culture it is 24 small When.It after 24 hours, is washed cell 2 times with PBS, and changes serum-free KBM (serum free keratinocyte basement into media).Ginsenoside Y is handled according to the concentration of the following table 19 respectively in each hole, and is reacted 30 minutes, is handled respectively later PGSA (10 μ g/ml), PGSA (50 μ g/ml) or PGSA (50 μ g/ml)+LPS (1 μ g/ml).Wherein, PGSA (peptidoglycan from S.aureus) is the peptide glycan (peptidoglycan) extracted from staphylococcus, well known Be PGSA be Gram-positive (+) bacterium cell wall primary structure, and the cell membrane component of bacterium can cause inflammation, especially Be allergic patients 90% can because staphylococcus causes superinfection, this is published content.In addition it is well known that rouge is more Sugared (LPS, lypopolysaccaride) is the main composition of the cell membrane of Gram-negative (-) bacterium, is to cause inflammation Main cause.

In 37 DEG C, 5%CO₂Incubator in cultivate 24 hours after, culture solution is taken, for interleukin 8 (Interleukin-8, IL-8) carries out ELISA, and the result is shown in the following table 19.ELISA uses manufacturing company (BD science) Experimental method.

Table 19

Component	1L-8 secretes (pg/ml)
		Untreated control group (Control)	935.12
PGSA(10μg/ml)	4812.60
		PGSA(50μg/ml)	5895.08
PGSA(50μg/ml)+LPS(1μg/ml)	6814.91
		Ginsenoside Y (5ppm)	1436.43
Ginsenoside Y (25ppm)	1204.19
		Ginsenoside Y (50ppm)	1012.73

It is capable of determining that from upper table 19, ginsenoside Y can be reduced significantly and be inhibited because PGSA and LPS are increased The secretion of IL-8.Therefore, Dermatologic preparation composition of the invention can be reduced significantly through the increased IL-8 of PGSA and LPS Secretion, so as to provide excellent anti-inflammatory effect.

Test example 15

Alleviate the evaluation of itch

Before experiment one day, by keratinocyte (cell strain title: HaCaT, at purchase: the ATCC) plant division on 96 orifice plates At 4 × 10⁴Cell number/hole, later in 37 DEG C, 5%CO₂Incubator in cultivate 24 hours.After 24 hours, balanced with Hank ' s Salting liquid (HBSS, Hank's Balanced Salt solution) washs 96 orifice plates twice, later by (2 μM of reaction buffer Fluo-4-AM, 20% pfpe acid (plurionic acid), 2.5mM 4- (dipropyl sulfamoyl) benzoic acid (probenecid)) it is added in cell.In 37 DEG C, 5%CO₂Incubator in react 30 minutes, at normal temperature react 30 points Zhong Hou is washed twice with HBSS buffer, and the ginsenoside Y of the concentration (%) in following table 20 handles cell.

Reaction after ten minutes, handles the trypsase (Trypsin) of 2U/ml or 5 μM of PAR-2 active peptide (SLIGKV), Intracellular Ca in measurement 80 seconds²⁺The variation of concentration.Intracellular Ca²⁺The measurement of concentration variation uses Flexstation3 (Flexstation3: multi-function microplate reader (Molecular Device), the U.S.).Handle 2U/ml trypsase or 5 μM PAR-2 active peptide calculates the difference of the contraction (flex) determined in 80 seconds and the maximum value and minimum value that obtain, by this later Difference is compared with the difference of minimum value and maximum value when the trypsase or 5 μM of PAR-2 active peptide of processing 2U/ml, Enter intracellular inhibiting rate (%) see the following table 20 for calcium ion.

Table 20

Be capable of determining that from upper table 20, the calcium ion of trypsase or PAR-2 active peptide enter intracellular amount according to The processing of ginsenoside Y and reduce, and with increase ginsenoside Y concentration, reduced significantly into intracellular calcium ion.

Therefore, the Dermatologic preparation composition of the invention containing ginsenoside Y, which can effectively inhibit, induces itch PAR-2 activity, so as to provide excellent antipruritic effect.

Dosage form example 4 and compare dosage form example 5

The composition of following table 21 has manufactured shampoo.Specifically, surfactant and ethylene glycol two are added in pure water Stearate, and 80 DEG C are heated to after equably dissolving, 40 DEG C are slowly cooled to by stirring, is thrown in said mixture Enter effective component according to the present invention and preservative, viscosity modifier, fragrance and hair conditioner and mixed, passes through stirring It is cooled to room temperature, to complete to manufacture.

Table 21

Test example 16

Reduce the effect test of dandruff

19~35 years old more 24 people of male of selected dandruff, is divided into two groups, every group of 12 people, and uses each dosage form respectively Example 4 and the shampoo for comparing dosage form example 5 use one month by the following method, measure dandruff slip later.

It before on-test, is had one's hair wash with common common shampoo, two days dandruffs, and comparative evaluation are accumulated in acquisition after hair washing The dandruff weight of above-mentioned acquisition and using each dosage form example 4 and compare washing primary for shampoo every two days and terminating to try for dosage form example 5 The weight of 2 days dandruffs is accumulated after testing.At this point, the dandruff of accumulation is directly acquired from scalp using vacuum absorption device, thus root Dandruff slip is found out according to following mathematical expressions 5, the result is shown in the following table 22.

Mathematical expression 5

Table 22

From upper table 22 it is found that the dosage form example 4 containing ginsenoside Y shows the effect of excellent anti-antidandruff.

Test example 17

Prevent the effect test of pruritus of scalp

Selected 24 people of more serious 25 years old~45 years old men and women of itching of the scalp, is divided into two groups, and every group of 12 people keep them every Passed through following evaluation criterions later using 2 weeks with the shampoo for comparing dosage form example 5 using primary each dosage form example 4 respectively within 3 days Evaluation prevents the effect of itching of the scalp, and the result is shown in the following table 23.

[evaluation criteria]

Table 23

Classification	Dosage form example 4	Compare dosage form example 5
			Pruritus of scalp eradicating efficacy	4	2.3

From upper table 23 it is found that the dosage form example 4 containing ginsenoside Y is to preventing itching of the scalp from showing more excellent effect.

Test example 18

Hair follicle hair papilla cell cultivation effect

The angling albumen for constituting hair is to form cell (keratinocyte) from hair root portion angling to generate, the angling shape It at cell is broken up from hair papilla cell.In order to evaluate this composition hair papilla cell activity, the present invention in use DP6 (rat immortalized dermal papilla cell) cell strain (Wendy Filsell, Journal of Cell Science 107,1761-1772(1994)).The strain of this hair papilla cell is the hair root from male PVG rat beard with micro- solution Cut open the cell strain that (microdissection) method is separated and cultivated, by the cell strain be added containing FBS (fetal calf serum, Fetal bovine serum) DMEM (Dulbecco's modified Eagle's medium, Gibco BRL, Gaithersburg, MD, the U.S.) in, and maintaining 5%CO₂, cultivate 24 hours in 37 DEG C of incubator.96 holes are added in DP6 In plate, and cultivate 24 hours in 37 DEG C of incubator, later by this ginsenoside Y be processed into respectively 5ppm, 10ppm and The concentration of 20ppm.After it's 24 hours pasts drug-treated, ability of cell proliferation is measured using WST-1 kit (Roche).As a result It see the table below 24.

Table 24

Classification	Ability of cell proliferation (%)
		Untreated control group (Control)	100
Ginsenoside Y (5ppm)	123
		Ginsenoside Y (10ppm)	131
Ginsenoside Y (20ppm)	146

It is able to confirm that out from upper table 24, when handling ginsenoside Y, will increase the proliferation of hair papilla cell, according to concentration There can be difference and significant increase.

Test example 19

Increase the effect assessment of potassium ion channel activity

Minoxidil as alopeciaing therapeutic agent is well known potential mitochondria K ~+Channel Opener (K_ATP Channel opener), it is the representative drugs for treating male sex hormone alopecia.In order to evaluate the machine of this minoxidil System uses following tests method: the processing blocking K from the fibroblast of corium for constituting scalp_ATPThe toluene sulphur fourth in channel Urea (SIGMA ALDRICH, T0891) is again turned on potassium-channel later and restores cell increasing to inhibit cell Proliferation It grows.

In order to evaluate this composition K ~+Channel Opener function, be employed herein as fibroblast NIH3T3 (Mouse embryonic fibroblast cell line) cell strain.This cell strain is will be small from NIH Switzerland The fibroblast 3T3 scheme (protocol) separated in mouse embryo (Swiss mouse embryo) carries out nature immortalization Cell strain.DMEM (Dulbecco's modified Eagle's medium, Gibco containing FBS is added in the cell strain BRL, Gaithersburg, MD, the U.S.) in, and maintaining 5%CO₂, cultivate 24 hours in 37 DEG C of incubator.By NIH3T3 Be added in 96 orifice plates, and cultivated 24 hours in 37 DEG C of incubator, post-processing orinase, after ten minutes will as sun The minoxidil of property control group is processed into 10 μM, and ginsenoside Y is processed into the concentration of 2.5ppm, 5ppm and 10ppm.At drug After managing 48 hours, ability of cell proliferation is measured using WST-1 kit (Roche).As a result 25 be see the table below.

Table 25

Classification	Ability of cell proliferation (%)
		Untreated control group (Control)	100
Minoxidil	132
		Ginsenoside Y (2.5ppm)	126
Ginsenoside Y (5ppm)	128
		Ginsenoside Y (10ppm)	133

From upper table 25 it is found that fibroblastic proliferation can be restored, and cell Proliferation energy when handling ginsenoside Y Power increases with the concentration of processed ginsenoside Y, and is able to confirm that, when ginsenoside Y is processed into 10ppm When, it is fibroblastic to be proliferated the level for reverting to minoxidil.

Test example 20

The effect test of ginsenoside Y promotion melanin production

The training adding 5% fetal calf serum, the benzyl penicillin of 100IU and 0.2 μM of TPA in RPMI culture medium and being formed Support in base, by melanocyte (melan-a) in 24 microwell plates (24-well microtiter plate) plant division at 50, 000 cells/well.Second day, ginsenoside Y is handled in the cell of plant division as substances, wherein the concentration of ginsenoside Y 100 μM of IBMX is handled as positive controls as the DMSO of negative control group processing 0.1% for 10ppm or 50ppm, It is cultivated 3 days under the conditions of 37 DEG C later.After culture, dissolved intracellular after the 1N NaOH of 100 μ l is added with PBS washing hole Melanin.The absorbance of the melanin through dissolving is measured in 405nm using microplate reader (microplate reader).Ginseng Saponin(e Y see the table below 26 to the result of the facilitation effect of melanin production compared with the control group.

Table 26

Sample	B16 cell amount (%)
		DMSO (0.1%)	100
IBMX(100μM)	120
		Ginsenoside Y (10ppm)	108
Ginsenoside Y (50ppm)	118

26 are seen the above table it is found that ginsenoside Y promotes the B16 cell of melanocyte, so that melanin production increases, because This shows the effect of excellent promotion melanin production.

Test example 21

Ginsenoside Y promotes the MITF in melanocyte and the effect of tyrosinase expression

Using 501mel cell strain, plant division is thin at 500,000 in 6 microwell plates (6-well microtiter plate) Born of the same parents/hole, as the DMSO of negative control group processing 0.1%, 100 μM of IBMX are handled as positive controls in each hole, Ginsenoside Y is processed into 10ppm as test group, and after cultivating 24 hours, 48 hours, 72 hours in 37 DEG C, obtained Protein.To thus obtained protein, western trace is carried out using MITF and tyrosinase antibody.By negative control The value of group, which is set to after the result after 100, western trace is compared with 100, to be shown in the following table 27.

Table 27

It sees the above table 27 to be able to confirm that out, ginsenoside Y can increase MITF and tyrosinase protein matter in melanocyte Expression.

Test example 22

The evaluation of the antibacterial ability of ginsenoside Y

In order to evaluate the antibacterial ability of ginsenoside Y, antibacterial experiment is implemented.Specific experimental method is as follows.

For the staphylococcus aureus (Staphylococcus aureus) of experiment, Escherichia coli (Escherichia Coli), Pseudomonas aeruginosa (Pseudomonas aeruginosa) bacterial strain is in pancreas protein soybeans culture medium (Tryptic Soy Broth it is cultivated in), and Candida albicans (Candida albicans), aspergillus niger (Aspergillus niger) bacterial strain It is cultivated in Sharpe fluid nutrient medium (Sabouraud's Dextrose Broth).By culture solution in each culture medium It is diluted to 1/100 (Candida albicans is 1/10), which is used as test bacteria liquid.Aspergillus niger be will manufacture at 2 × 10⁸The spore suspension of cfu/ml is used as test bacteria liquid.

It is molten to be used as dilution by the test bacteria liquid that 0.15ml is added in each culture medium of 15ml for uniformly mixed solution Liquid.

The sample of 16 μ l and the dilute solution of 184 μ l are added in the first row of 96 orifice plates (96well plate).Remaining The dilute solution of 100 μ l is added in hole.After mixing by the mixed liquor of the first row, 100 μ l is taken to be added in the second row and mix It closes uniformly, takes 100 μ l to be added in the third line later, diluted respectively twice by the above method.

Staphylococcus aureus (Staphylococcus aureus), Escherichia coli (Escherichia coli), green pus Bacillus (Pseudomonas aeruginosa) is cultivated in 32 DEG C of thermostat, and Candida albicans (Candida Albicans), aspergillus niger (Aspergillus niger) is cultivated in 25 DEG C of thermostat.

After 48 hours, whether confirming the proliferation of bacterium by suspension degree and microscope, and minimum inhibitory concentration is determined (MIC), the result is shown in the following table 28.

Table 28

Shown in table 28 as above, confirm that ginsenoside Y shows antibacterial ability to a variety of bacterial strains, what it is thus, it is possible to prediction is Ginsenoside Y can be used as natural antiseptic agent or antibacterial agent in composition.

Claims (4)

1. a kind of Dermatologic preparation composition containing the ginsenoside Y as principle active component is in preparation for improving acne Cosmetic composition in application.

2. the change that a kind of Dermatologic preparation composition containing the ginsenoside Y as principle active component is used for antibacterial in preparation Application in cosmetic compositions.

3. a kind of Dermatologic preparation composition containing the ginsenoside Y as principle active component is in preparation for inhibiting lipid Application in the cosmetic composition of generation.

4. a kind of Dermatologic preparation composition containing the ginsenoside Y as principle active component is in preparation for improving allergy Cosmetic composition in application.