KR20190059058A - Composition comprising camellia sinensis L. root extract for Enhancing Skin Barrier - Google Patents

Abstract

(NM_002963), IL-36G (NM_019618), and LCE3D (NM_032563), which are expressed in the skin cells, in which the expression level is affected by a stimulus source causing skin barrier weakness, A composition for enhancing skin barrier that regulates at least one expression level selected from the group consisting of < RTI ID = 0.0 > a < / RTI >

Description

[0001] The present invention relates to a composition for enhancing skin barrier comprising root extract of tea root (L. root extract for enhancing skin barrier)

Disclosed herein are compositions for enhancing skin barrier. More specifically, a composition comprising a tea root extract which strengthens a skin barrier by significantly changing a biomarker such as a skin cell gene whose expression level is changed by weakening skin barrier as compared with a normal skin cell is disclosed do.

Among the constitutive layers of the skin, the epidermis plays an important role in preventing water evaporation inside the human body. The epidermis is divided into stratum corneum, granular layer, superficial layer, and basal layer in order from the outside. Cells of stratum corneum act like bricks, and interstitial lipids between keratinocytes act like mortar to form skin barrier. In addition, a healthy human keratinocyte has a high concentration of natural moisturizing factor (NMF) to help maintain moisture in the skin. For example, substances such as amino acids are water-soluble, Thereby suppressing drying.

However, as it is nowadays, there is a tendency to adjust the temperature of the air / heating due to the change of environment or life pattern, the stress caused by various stresses and environmental pollution in the social life, frequent washing according to make- Due to various causes such as aging of the skin, the skin becomes dry, the surface becomes rough, the skin becomes loose, the moisture is lost, and the appearance of the skin does not appear, and thus the need for a skin moisturizer increases . Excessive physical and chemical stimuli, ultraviolet rays, stress and nutrient deficiency, which are externally received, can lower the normal functions of the skin and promote elasticity loss, keratinization and wrinkle formation. Especially, . Excessive physical, chemical stimuli, ultraviolet rays, and stress from the outside of the skin lower the normal functions of the skin and promote skin aging phenomenon such as loss of elasticity, keratinization, and wrinkle formation. Especially, .

It is known that IL-36G is a useful biomarker in psoriasis caused by skin barrier weakness (see Non-Patent Document 1). S100A7 and S100A8 are also known to be indicators of atopic dermatitis and psoriasis due to impaired skin barrier function. In particular, an increase in the expression level of S100A8 is related to the activation state of keratinocytes, and it is disclosed that it is not caused by the accompanying inflammation (see Non-Patent Document 2), which distinguishes it from the inflammation phenomenon in the skin.

 AM D'Erme, " IL-36c (IL-lF9) Is a Biomarker for Psoriasis Skin Lesions ", Journal of Investigative Dermatology, Volume 135, 2015.  Eckert et al, " S100 Proteins in the Epidermis ", J Invest Dermatol, 123 (1): 23-33, 2004 Jul.

Thus, the present inventors have found that a stimulus source that causes skin barrier weakness has a detrimental effect on the skin, and by such influence, affects the expression of a skin cell gene, thereby causing a symptom such as a stimulation source that causes skin barrier weakness. ≪ / RTI > have the effect of potentiating the skin barrier.

Therefore, in one aspect, the present invention aims to provide a composition for skin barrier enhancement.

In order to achieve the above object, in one aspect, the present invention provides a composition comprising an extract of a tea plant root as an active ingredient, wherein the expression level of the IL-36G gene, which is an intracellular gene that is affected by a stimulus source causing skin barrier weakness Wherein at least one expression level selected from the group consisting of S100A7 (NM_019618), S100A7 (NM_002963), and S100A8 (NM_002964) is regulated to a normal level.

In one aspect, by using the composition for skin barrier enhancement provided by the present invention, it is possible to reduce the damage of skin cells by returning the amount of gene expression, which is changed by a stimulus source causing skin barrier weakness, to a normal level.

1 shows the cell survival rate by stimulus treatment, wherein ADSP represents Asian dust-storm particles, and PM10 (Particulate matter 10) represents fine dust particles having a particle size of 10 μm And PM 2.5 (Particulate matter 2.5) represent fine dust particles having a particle size of 2.5 μm.
FIG. 2A shows that the expression level of IL-36G gene changed in skin cells to which a stimulus causing skin barrier weakness was applied was returned to a normal level by treatment with a tea root extract.
FIG. 2B shows that the expression level of the S100A7 gene changed in the skin cells to which the stimulus causing the weakening of the skin barrier was applied was returned to the normal level by the treatment of the tea root extract.
FIG. 2C shows that the expression level of the S100A8 gene changed in the skin cells to which the stimulus causing the weakening of the skin barrier was applied was returned to the normal level by the treatment of the tea root extract.
In the figure, " Rope 1 " indicates that the ethanol extract of tea root (Example 1) is treated at 1 ppm, " Rope 5 " is the result of treating 5 ppm of the tea root ethanol extract (Example 1) (Example 2) treated with 1 ppm of the ethanol extract of the tea root roots, 5 ppm of the butanol fraction of the tea root roots ethanol extract (Example 2), " NT " , &Quot; cont1 " and " cont2 " mean respective control groups.

Hereinafter, the present invention will be described in detail.

In one aspect of the present invention, the composition may comprise a tea root root extract as an active ingredient.

Camellia sinensis is a perennial evergreen plant belonging to theaceae. It is distributed in tropical, subtropical and temperate regions. The main ingredients of tea tree are catechins, saponins, caffeine, amino acids, vitamins and minerals. Have been reported to exhibit various physiological activities and pharmacological effects. Today, it has been actively studied as a physiologically active substance contributing to health.

In one aspect, the tea root extract may be an extract comprising saponin.

In one aspect, the tea root extract may contain saponin in an amount of 30 to 70% by weight based on the total weight of the extract. Preferably, the tea root root extract may contain saponin in an amount of 50 to 60% by weight based on the total weight of the extract. When the content of saponin was within the above range, the anti-aging and anti-inflammatory effect by the tea root extract was excellent.

More specifically, it is preferable that it is 30 wt% or more, 31 wt% or more, 32 wt% or more, 33 wt% or more, 34 wt% or more, 35 wt% or more, 36 wt% or more, 37 wt% or more, 38 wt% At least 40 wt%, at least 41 wt%, at least 42 wt%, at least 43 wt%, at least 44 wt%, at least 45 wt%, at least 46 wt%, at least 47 wt%, at least 48 wt% At least 50 wt.%, At least 51 wt.%, At least 52 wt.%, At least 53 wt.%, At least 54 wt.%, Or at least 55 wt.% Of at least 70 wt.%, , 67 weight% or less, 66 weight% or less, 65 weight% or less, 64 weight% or less, 63 weight% or less, 62 weight% or less, 61 weight% or less, 60 weight% or less, 59 weight% or less, , 57 wt% or less, 56 wt% or less, or 55 wt% or less, but is not limited thereto.

In one aspect of the present invention, the tea root can be extracted with a specific extraction solvent to form a tea root extract.

In one aspect of the present invention, the tea root extract can be prepared by extracting tea roots with water or a primary extraction solvent which is an organic solvent. Specifically, the primary extraction solvent is selected from the group consisting of water, C ₁ -C ₆ anhydrous or lower alcohol, acetone, butylene glycol, ethyl acetate, diethyl acetate, diethyl ether, benzene, chloroform and hexane Any one or more can be daily for extraction. More particularly, the extraction comprises a polar solvent comprising water, anhydrous or lower alcohol of C ₁ -C ₆ , acetone and butylene glycol, and ethyl acetate, diethyl acetate, diethyl ether, benzene, chloroform and hexane Or a low-polarity solvent which is a solvent for the solvent. More specifically, the solvent may be 50-90% ethanol aqueous solution, and may be 60-80% ethanol or 65-75% ethanol aqueous solution. When the solvent is 50-90% aqueous ethanol solution, the active ingredient can be effectively extracted from the tea root. In one embodiment, the solvent may be about 70% aqueous ethanol solution.

In one aspect, the tea root extract can be prepared by extracting with the primary extraction solvent and fractionating with a secondary extraction solvent. Specifically, the secondary extraction solvent may be anhydrous or functional butanol. The content of the specific component contained in the extract can be further increased through the fraction, and the content of the saponin contained in the tea root extract can be further increased. For example, the saponin content of the extract obtained by fractionating the tea root root extract with the secondary extracting solvent from the tea root extract extracted with the primary extracting solvent is, for example, not less than 5% by weight, not less than 6% by weight , 7% or more, 8% or more, 9% or more, 10% or more, 11% or more, 12% or more, 13% or more, 14% or more, or 15% And may be increased by 5-15% by weight.

In one embodiment, the tea root extract is obtained by extracting 70% ethanol with a primary extraction solvent and butanol with a secondary extraction solvent. For example, the saponin content of the tea root extract extracted with the 70% ethanol as the first extraction solvent may be about 55.8% by weight based on the total extract weight, and the tea root extract may be fractionated with butanol as a second extraction solvent The saponin content of one extract may be about 67.6% by weight based on the total weight of the extract. Thus, the saponin content of the extract through the fraction can be increased by about 11.8% by weight.

In one aspect, the tea root extract may be obtained by extracting with the extraction solvent, followed by addition of one or more of filtration, concentration, separation or drying. In particular, the tea root extract may be subjected to one or more filtration processes.

In one aspect, the extract can be concentrated under reduced pressure to a suitable temperature in a distillation apparatus equipped with a cooling condenser after extraction, specifically to a temperature of about 50 캜.

However, the tea root extract of the present invention can be obtained by extracting by conventional methods in the art, and is not limited by the above-mentioned method.

In one aspect of the present invention, the composition may comprise from 0.000001 to 40% by weight of the tea root root extract, based on the total weight of the composition. When the content of the extract is 0.000001 to 40% by weight, the skin care effect by fine dust caused by the composition containing the tea root extract is excellent.

More specifically, 0.0000001 wt% or more, 0.0000005 wt% or more, 0.0000007 wt% or more, 0.0000009 wt% or more, 0.0000009 wt% or more, 0.000001 wt% or more, 0.000002 wt% or more, 0.000004 wt% or more, 0.000006 wt% or more, 0.000008 wt. At least 0.00003% by weight, at least 0.00005% by weight, at least 0.00007% by weight, at least 0.00009% by weight, at least 0.00009% by weight, at least 0.0001% by weight, at least 0.0003% by weight, at least 0.0005% by weight, at least 0.0007% by weight, % Or more, 0.01 wt% or more, 0.1 wt% or more, 1 wt% or more, 3 wt% or more, 5 wt% or more, 7 wt% or more, 9 wt% or more, 10 wt% or more, 13 wt% or more, At least 17% by weight, at least 19% by weight, at least 21% by weight, at least 23% by weight, at least 25% by weight, at least 27% by weight, at least 29% by weight, at least 30% by weight, at least 31% At least 33 wt%, at least 34 wt%, at least 35 wt%, at least 36 wt%, at least 37 wt%, at least 38 wt% 39 wt% or less, 38 wt% or less, 37 wt% or less, 36 wt% or less, 35 wt% or less, 34 wt% or less, 33 wt% or less, 32 wt% or less % Or less, 31 wt% or less, 30 wt% or less, 29 wt% or less, 28 wt% or less, 26 wt% or less, 24 wt% or less, 22 wt% or less, 20 wt% or less, 18 wt% Up to 14 wt.%, Up to 12 wt.%, Up to 10 wt.%, Up to 9 wt.%, Up to 8 wt.%, Up to 6 wt.%, Up to 4 wt.%, Up to 2 wt. %, 0.09 wt% or less, 0.04 wt% or less, 0.01 wt% or less, 0.006 wt% or less, 0.001 wt% or less, 0.0009 wt% or less, 0.0007 wt% or less, 0.00005 wt% or less, 0.00003 wt% %, 0.000009 wt% or less, 0.000007 wt% or less, 0.000005 wt% or less, 0.000003 wt% or less, 0.000001 wt% or less, 0.0000009 wt% or less, 0.0000007 wt% or less, 0.0000005 wt% % Or less, but can be less than 0.0000003% by weight or less, 0.0000002% by weight or less, 0.0000001% by weight or less, or 0.00000009% by weight, but is not limited thereto.

Another aspect of the present invention may include the use of the composition of the present invention to enhance skin barrier. Specifically, the composition may be a composition for enhancing skin barrier comprising an extract of Root Root as an active ingredient.

In one aspect, the present invention provides a composition for enhancing skin barrier by regulating the expression level of a specific gene in skin cells damaged by a stimulus causing skin barrier weakness to a normal level.

Specifically, IL-36G (NM_019618), S100A7 (NM_002963), and S100A8 (NM_002964) are examples of genes in the skin cells to which the expression level is affected by stimulation that causes skin barrier weakness in the present invention. Since the expression levels of IL-36G (NM_019618), S100A7 (NM_002963) and S100A8 (NM_002964) are increased by stimulation that causes skin barrier weakness, the expression level of these genes is suppressed and regulated to a normal level .

In one aspect, the genes whose expression levels are increased by the stimuli causing skin barrier weakness used in the present invention are shown in [Table 1]. In the table, Name means the genebank accession ID of NCBI, Gene Symbol means the official gene symbol, and Gene title means the name of each gene.

Increase gene Name Gene
Symbol Gene title NM_019618 IL36G interleukin 36, gamma NM_002963 S100A7 S100 calcium binding protein A7 NM_002964 S100A8 S100 calcium binding protein A8

Analysis of the expression level of the gene or protein can be performed using a microarray, PCR, NGS (Nest Generation Sequencing), Western blot, northern blot, ELISA, radioimmunoassay, radioimmunoassay, tissue immuno staining, Can be analyzed using a variety of analytical methods known in the art.

In one aspect of the present invention, the composition may be a cosmetic composition, a pharmaceutical composition, or a health functional food composition.

Examples of the cosmetic composition include cosmetics such as various creams, lotion creams, lotions, skins, and the like, and cleansing, cleansing agents, soaps, and essences.

In one aspect, the cosmetic composition to which the composition containing the tea root extract of the present invention is added may take the form of a solution, an emulsion, a viscous mixture or the like.

That is, the cosmetic as one aspect of the present invention is not particularly limited in its formulation and may be, for example, an emulsion, cream, lotion, essence, pack, gel, powder, makeup base, foundation, Formulations such as foam, cleansing cream, cleansing water, body lotion, body cream, body oil, body essence, shampoo, rinse, body cleanser, soap, hair dye, spray and the like.

In the cosmetic composition of each formulation, the components other than the tea root extract may be mixed and selected without difficulty by those skilled in the art depending on the formulation or purpose of use of other cosmetics.

The cosmetic composition according to one aspect of the present invention may further comprise a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymeric polysaccharides, sphingolipids and seaweed extracts.

In one aspect, the cosmetic composition of the present invention may contain, in addition to the above essential ingredients, other ingredients usually added to cosmetics, if necessary.

Examples of the compounding ingredients that may be added include organic solvents such as a preservative component, a moisturizer, an emollient, a surfactant, an organic and inorganic pigment, an organic powder, an ultraviolet absorbent, a preservative, a bactericide, an antioxidant, a plant extract, a pH adjuster, A blood circulation accelerator, a cold agent, an antiperspirant agent, and purified water.

The components to be added in addition to these components are not limited thereto, and any of the above components can be compounded within a range that does not impair the objects and effects of the present invention.

In one aspect, the pharmaceutical composition comprising the tea root extract of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions.

As the formulation, the pharmaceutical composition containing the tea root extract may be formulated into tablets, capsules, powders or syrups, or external preparations such as ointments, gels, creams, patches or sprays, And can be formulated and used in any form suitable for pharmaceutical preparations.

In general, it is to be understood that the actual dosage of the active ingredient administered by the pharmaceutical composition should be determined in light of various relevant factors such as the severity of the symptoms, the route of administration selected, the age, sex, weight and health status of the subject. In general, the dosage of the active ingredient may be from 0.0001 mg / kg / day to 3000 mg / kg / day, for example from 10 mg / kg / day to 500 mg / kg / day.

In the health functional food composition as one aspect of the present invention, the health food is produced by using a raw material or a component (functional raw material) having a function useful for a nutrient or a human body which is likely to be deficient in a daily meal, Or to maintain or improve health through the activation of physiological functions. However, the present invention is not limited thereto. The health food may be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, and circles, but is not limited thereto and may be manufactured and processed in any form according to the law.

Specifically, the health beverage composition has no particular limitation on the other ingredients other than the above-mentioned compounds as essential ingredients in the indicated ratios, and may contain various flavors or natural carbohydrates as additional ingredients such as ordinary beverages. Examples of natural carbohydrates are conventional sugars such as monosaccharide polysaccharides, cyclodextrins and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavors (tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be used as flavorings other than those described above.

In general, it should be understood that the actual dosage of the active ingredient administered by the health functional food composition should be determined in light of various relevant factors such as the severity of the symptoms, the selected route of administration, the age, sex, weight and health status of the subject . In general, the dosage of the active ingredient may be from 0.0001 mg / kg / day to 1000 mg / kg / day, for example from 0.02 mg / kg / day to 6 mg / kg / day.

Hereinafter, the configuration and effects of the present invention will be described in more detail with reference to examples. However, these examples are provided for illustrative purposes only in order to facilitate understanding of the present invention, and the scope and scope of the present invention are not limited by the following examples.

[Example 1] Production of ethanol extract of tea root roots

The roots of Camellia sinensis L. obtained from Jeju Oshulok Ranch were washed with purified water and dried. Then, 100 g of the tea root powder obtained by cementation was added to 1 L of 70% ethanol aqueous solution, and the mixture was stirred at room temperature for 12 hours or longer. Filter 2 with filter paper. The extract thus obtained was concentrated under reduced pressure at 50 ° C using a distillation apparatus equipped with a cooling condenser and dried to obtain a 70% ethanol extract of tea roots (dry weight: 21.05 g). The saponin content of the tea root 70% ethanol extract obtained by the above procedure was about 55.8% by weight based on the total weight of the extract.

[Example 2] Production of butanol fraction of ethanol extract of tea root roots

10 g of the extract obtained in Example 1 was dissolved in 200 mL of distilled water, and 1 L of the extract was added to a separatory funnel, 200 mL of butanol was added thereto, and the mixture was shaken thoroughly. Then, the upper layer (butanol layer) was completely separated into two layers. The lower layer (water layer) is extracted by repeating the above procedure twice more by using a separatory funnel. The separated upper layers were combined and concentrated under reduced pressure at 50 ° C using a distillation apparatus equipped with a cooling condenser and dried to obtain a butanol fraction (dry weight: 4.83 g) of the ethanol extract of tea root root. The saponin content of the butanol fraction of the tea root ethanol extract obtained by the above procedure was about 67.6% by weight based on the total weight of the extract.

[Example 3] Preparation of skin barrier weakening stimulus source

The filter pack was used for about 24 hours by replacing the filter and denuder at around 10:00 am on each measurement day. The filter pack and the denuder were replaced by a low volume air sampler (Sensidyne, Gillian, Low Volume Air Sampler, FL, USA) Respectively. For 28 days, fine dust was collected daily from the windy area of Seoul (Gyeonggi-do, Korea), and the measurement time was measured by turning on the timer while turning on the vacuum pump and turning off the timer Time was recorded. The flow rate was measured with a flow meter (Model 4143, TSI Inc.) at the start of the measurement at a flow rate of 16.7 L / min, and the flow rate was measured again at the end of the measurement. The Teflon filter in the filter pack weighed before and after sampling. Before measuring the weight of the Teflon filter, the sample was weighed into a desiccator (NIKKO, Japan) with a relative humidity of 40% for 24 hours and then weighed twice on an electronic balance (DVG215CD, Ohaus) . After the sample was collected, the sample was weighed in a desiccator for 24 hours before measuring the weight, and then the weight was measured twice, and the mass concentration was calculated by comparing with the weight measured before the sampling. The extraction of fine dust was carried out by wetting the Teflon filter with 1 mL of ethanol, placing the filter with 14 mL of DW, closing the lid with the filter surface of the filter touching the water surface, and ultrasonically applying the filter with the ultrasonic cleaner for 30 minutes. In order to minimize the error caused by moisture in the filtration step, the water content of the filter was completely removed for 48 hours in a decicator, and then the weight of the filter was measured using a super-precision scale system (Mettler Toledo Company) Weighed and weighed before and after filter extraction.

[Example 4] Culture of keratinocyte (normal human) cell line

Human normal epidermal keratinocytes were purchased from Lonza, Inc. (Walkersville, Maryland, USA), subcultured and cultured in a CO 2 incubator at 37 ° C under 5% CO 2 Lt; / RTI > The cell culture was in accordance with Lonza's guidelines. (KGM-2 Bullet kit, CC-4152) (ingredient: BPE (bovine pituitary extract)), human epidermal growths (KBM- (Gentamycin Suflate + Amphofericin-B: GA), human epidermal growth factor (hEGF), insulin, Hydrocortisone, Transferrin, Epinephrine and gentamycin sulfate (KGM-2 Bullet Kit, CC-3107) was added to the reaction mixture.

[Example 5] (Normal human) Treatment of a stimulus source and measurement of cytotoxicity on a keratinocyte cell line

MTT experiments were carried out using keratinocyte lines (normal human) by the method of Mossman et al. (J. Immunol. Methods, 65, 55-63, 1983) in order to confirm cytotoxicity through fine dusting.

Specifically, using a 24-well plate, and the cell culture conditions of Example 3 taken to prepare a dispersion of fine particles by dispersing the fine particles in the obtained diameter 2.5㎛ in purified water in the following Example 4 2.5 × 10 ⁵ Cells cultured under the conditions of the number of well cells were treated with the fine dust dispersion and cultured for 24 hours. Then, 5 mg / ml of MTT (3-4,5-dimethylthiazol-2,5-diphenyltetra zolium bromide) Lt; 0 > C for 3 hours. The medium was then removed and the formazan crystal formed was dissolved in 500 占 퐇 of DMSO. The lysate was transferred to a 96-well plate and aliquoted and the OD value measured at 540 nm absorbance. The measurement results are shown in Fig.

As shown in FIG. 1, the concentration (IC20) showing an 80% survival rate with respect to cytotoxicity by a dispersion in which fine particles of 2.5 micrometer or less were dispersed in the cell line was 12.5 g / Ml.

[Example 6] Confirmation of cell gene change by stimulation causing skin barrier weakness through next generation sequencing

For RNA-base sequence data processing and analysis, reference was made to the general analysis step developed by Trapnell et al. (2012). We confirmed the RNA-seq data quality using FastQC (http://www.bioinformatics.babraham.ac.uk/projects/fastqc/) and used FASTX (http://hannonlab.cshl.edu/fastx_toolkit/) , Thereby removing the base and the adder sequences with low accuracy. Afterwards, alignment was performed using Tophat (Trapnell et al., 2009) and human genome (hg19), and the amount of data for each sample was confirmed using EVER-seq renamed RSeQC (Wang et al., 2012 ). In addition, the level of expression of transcripts was quantified using Cufflinks, and transcription levels were compared between the fine dust dispersion treated and normal samples (Trapnell et al., 2010). A stringent cutoff of ≥2.0 fold-change was applied to the FDR adjusted p-value <0.05 to determine the gene that showed significant expression differences in the treatment of the dispersion of fine dust with a diameter of 2.5 μm. The measurement results are shown in Figs. 2A to 2C below.

[Example 7] Real-time RT-PCR quantification

Human microkeratin skin cells cultured in Example 4 in the amount of fine particles having a diameter of 2.5 탆 extracted in Example 3 were treated with 1 ml of the cell culture medium in an amount of 12.5 占 퐂 and applied to the applied biosystems TaqMan® Primers) to determine the relative mRNA expression levels. The tea root extract and the butanol fraction used in Examples 1 and 2 were used.

Name Gene
Symbol TaqMan® primer NM_019618 IL36G Hs00219742_m1 NM_002963 S100A7 Hs00161488_m1 NM_032563 LCE3D Hs00754375_s1

After 24 hours, the culture medium was removed, and the cells were washed with 2 ml of phosphate buffered saline (PBS). Then, the cells were washed with 1 ml of phosphate buffered saline (PBS) Trizol reagent (Invitrogen, Carlsbad, CA, USA) was used to isolate the intracellular RNA. The separated RNA was further purified with an RNA kit (QIAGEN RNeasykt, QIAGEN, Valencia, Calif.) From Agilent Technologies, Inc., Agilent 2100 BioAnalyzer, Agilent Technologies, Santa Clara, ) Was used to confirm the quality of the RNA. CDNA was synthesized from the RNA using a reverse transcription kit (Invitrogen, Carlsbad, Calif.) Of Invitrogen, and the cDNA was synthesized using the primers in the real-time RT-PCR -RT-PCR: real-time reverse transcription polymerase chain reaction). The change in gene expression pattern was evaluated by real-time PCR using a TaqMan gene expression assay kit (Applied Biosystems, Foster City, Calif.) Of Applied Biosystems. As shown in Fig. 2C. Both Q-RT-PCR and real-time PCR were performed according to the standard protocols distributed by Life Technologies. Specifically, the Q-RT-PCR was performed at 95 ° C for 20 seconds, followed by 95 ° C for 3 seconds and 60 ° C for 30 seconds For 40 cycles.

As shown in FIGS. 2A to 2C, there is a gene whose expression level is increased in skin cells stimulated by a stimulus that causes skin barrier weakness. In the case of interleukin-36 gamma (IL-36G ), S100 calcium-binding protein A7 (S100A7), and S100 calcium-binding protein A8 (S100A8) gene.

Therefore, the tea root extract effectively inhibits skin cells from skin cell damage caused by stimulation that causes skin barrier weakness, inhibits or prevents the change in the expression level of the specific gene described above by stimulation that causes skin barrier weakness, Lt; RTI ID = 0.0 &gt; of &lt; / RTI &gt;

Hereinafter, formulation examples of the composition according to the present invention will be described. However, the cosmetic composition, the pharmaceutical composition and the health functional food composition can be applied to various formulations, and the present invention is not limited thereto .

[Formulation Example 1] Tablets

100 mg of tea root extract, 400 mg of lactose, 400 mg of corn starch and 2 mg of magnesium stearate according to the examples of the present invention were mixed and tablets were prepared by tableting according to a conventional preparation method of tablets.

Compounding ingredient Content (mg) Tea root extract 100 Lactose 400 Corn starch 400 Magnesium stearate 2

[Formulation Example 2]

100 mg of tea root extract, 400 mg of lactose, 400 mg of corn starch and 2 mg of magnesium stearate according to the present invention were mixed and filled in gelatin capsules according to the conventional preparation method of capsules to prepare capsules.

Compounding ingredient Content (mg) Tea root extract 100 Lactose 400 Corn starch 400 Magnesium stearate 2

[Formulation Example 3]

50 mg of tea root extract, 250 mg of anhydrous crystalline glucose and 550 mg of starch according to the present invention were mixed and granulated into granules using a fluidized bed granulator.

Compounding ingredient Content (mg) Tea root extract 50 Anhydrous crystalline glucose 250 Starch 550

[Formulation Example 4] Soap

Compounding ingredient content(%) Tea root extract 5.00 maintain Suitable amount Sodium hydroxide Suitable amount Sodium chloride Suitable amount Spices Suitable amount Purified water Balance

[Formulation Example 5] Lotion

Compounding ingredient content(%) Tea root extract 5.00 L-ascorbic acid-2-phosphate magnesium salt 1.00 Water soluble collagen (1% aqueous solution) 1.00 Sodium citrate 0.10 Citric acid 0.05 Licorice extract 0.20 1,3-butylene glycol 3.00 Purified water Balance

[Formulation Example 6] Cream

Compounding ingredient content(%) Tea root extract 3.00 Polyethylene glycol monostearate 2.00 Self emulsifying monostearic acid glycerin 5.00 Cetyl alcohol 4.00 Squalene 6.00 Tri-2-ethylhexanoic acid glyceryl 6.00 Sphingoglycolipids 1.00 1,3-butylene glycol 7.00 Purified water Balance

[Formulation Example 7] ointment

Compounding ingredient content(%) Tea root extract 5.00 Polyvinyl alcohol 13.00 L-ascorbic acid-2-phosphate magnesium salt 1.00 Lauroylhydroxyproline 1.00 Water soluble collagen (1% aqueous solution) 2.00 1,3-butylene glycol 3.00 ethanol 5.00 Purified water Balance

 [Formulation Example 8] Preparation of serum

Compounding ingredient content(%) Tea root extract 3.00 Hydroxyethylene cellulose (2% aqueous solution) 12.00 Xanthan gum (2% aqueous solution) 2.00 1,3-butylene glycol 6.00 Concentrated glycerin 4.00 Sodium hyaluronate (1% aqueous solution) 2.00 Purified water Balance

[Formulation Example 9] Health food

Compounding ingredient content Tea root extract 2 mg Vitamin A acetate 70 μg Vitamin E 1.0 mg Vitamin B1 0.13 mg Vitamin B2 0.15 mg Vitamin B6 0.5 mg Vitamin B12 0.2 μg Vitamin C 10 mg Biotin 10 μg Nicotinic acid amide 1.7 mg Folic acid 50 μg Calcium pantothenate 0.5 mg Ferrous sulfate 1.75 mg Zinc oxide 0.82 mg Magnesium carbonate 25.3 mg Potassium monophosphate 15 mg Dicalcium phosphate 55 mg Potassium citrate 90 mg Calcium carbonate 100 mg Magnesium chloride 24.8 mg

[Formulation Example 10] Health drinks

Compounding ingredient content Tea root extract 50 mg Citric acid 1000 mg oligosaccharide 100 g Taurine 1g Purified water Balance

Claims (12)

A composition for enhancing skin barrier comprising an extract of tea root as an active ingredient. The method according to claim 1,
Wherein the tea root extract is an extract comprising saponin. 3. The method of claim 2,
Wherein the tea root extract comprises 30 to 70% by weight saponin based on the total weight of the extract. The method according to claim 1,
Wherein the tea root extract is selected from the group consisting of water, C ₁ -C ₆ anhydrous or lower alcohol, acetone, butylene glycol, ethyl acetate, diethyl acetate, diethyl ether, benzene, chloroform and hexane Which is extracted with a tea extracting solvent and fractionated into a second extracting solvent which is anhydrous or functional butanol. 5. The method of claim 4,
Wherein the tea root extract is firstly extracted with anhydrous or hydrated ethanol and fractionated with anhydrous or functional butanol. The method according to claim 1,
Wherein the tea root extract is included in an amount of 0.000001 to 40% by weight based on the total weight of the composition. The method according to claim 1,
Wherein said composition inhibits expression of at least one selected from the group consisting of IL-36G (NM_019618), S100A7 (NM_002963) and S100A8 (NM_002964). 8. The method of claim 7,
Wherein the composition is applied to keratinocytes. The method according to claim 1,
Wherein the tea root extract is administered at a dose of 10 to 500 mg / kg / day. The method according to claim 1,
Wherein the composition is a cosmetic composition. The method according to claim 1,
Wherein the composition is a pharmaceutical composition. The method according to claim 1,
Wherein the composition is a health functional food composition.

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