TW201904551A - Contains a composition of MENTHA ARVENSIS extract for the treatment of micro-dust (MICRODUST) damage to skin cells, enhancement of skin barriers, and anti-aging or anti-inflammatory - Google Patents

Abstract

The present disclosure discloses a composition for external application to the skin for caring skin damages by microdust, comprising Mentha arvensis extract as an active ingredient, and which regulates the expression level of at least one selected from the group consisting of IL-1a (NM_000575), IL-1B (NM_000576), IL-36G (NM_019618), S100A7 (NM_002963), LCE3D (NM_032563), PTGS2 (NM_000963), and XDH (NM_000379), which are genes in skin cells whose expression levels are affected by microdust, to the normal level. It is possible to return the gene expression levels changed by microdust to the normal levels and thereby to care damages of skin cells, by using the composition for caring skin damages by microdust.

Description

Composition containing MENTHA ARVENSIS extract, used to treat microdust damage to skin cells, strengthen skin barriers, and anti-aging or anti-inflammatory

The present invention relates to a composition for caring for the damage of fine dust on skin cells, especially a composition containing wild mint extract, and the composition takes care of the damage of skin cells by significantly changing biomarkers etc. It is a gene of skin cells whose expression level is changed by fine dust compared with the normal state.

The skin is a part of the body and is directly exposed to the external environment. It not only serves as a protective layer to protect our vital organs, but also regulates the evaporation of water and protects the body from external infections. However, even if the skin is to prevent viruses from invading from the outside, excessive skin exposure to ultraviolet rays or contaminants can cause skin irritation. In particular, the skin is damaged by Asian dust accompanied by strong winds and dirt.

Asian dust is a phenomenon in which small sand or red clay floats from inland deserts such as China and Mongolia, is taken away by the wind and then falls near the ground. In South Korea, Asian dust regularly occurs every spring. Asian dust is a combination of organic and inorganic materials, and its physical properties and composition are very diverse according to the time and place of occurrence. It also includes metals that can produce biological effects. Large particles of Asian dust often remain in their source or surrounding environment, while small particles even flow into South Korea. It is reported that when inhaled, this dust deposits in the lower bronchus and even in the gas exchange part of the lungs, which may cause damage to the respiratory system. In addition, it was found that people living in dusty areas or areas with a lot of Asian dust have increased skin cell damage in their skin.

At the same time, in the skin layer, the epidermis plays an important role in preventing water evaporation in the human body. From the outside, the epidermis layer is: stratum corneum, stratum granulosum, stratum spinosum, and stratum basale. The cells of the stratum corneum act like bricks, and the intercellular lipids between the stratum corneum cells form a skin barrier like mortar. In addition, healthy people's stratum corneum cells have a high concentration of natural moisturzing factor (NMF), which helps maintain moisture in the skin. For example, substances such as amino acids are water-soluble and therefore effectively combine with moisture to prevent skin moisture from drying out.

However, nowadays, due to various reasons, such as artificial temperature control of cooling / heating due to changes in the environment or lifestyle, skin pressure due to the pressure of social life and environmental pollution, frequent washing due to makeup habits, and age increase Of natural skin aging, the cuticle moisture is reduced, so that the skin becomes dry, the skin surface becomes rough, and the skin becomes loose and looks discolored due to lack of moisture, etc. Therefore, the demand for skin moisturizers has increased. In addition, excessive physical and chemical irritation from outside, ultraviolet rays, stress, and nutritional deficiencies degrade the normal function of the skin and promote phenomena such as loss of elasticity, keratinization, and wrinkle formation. Specifically, the dermis-epidermis boundary is severely damaged by ultraviolet light.

IL-36G is known as a biomarker useful in psoriasis caused by skin barrier weakness (see Non-Patent Document 1). S100A7 is also known as an indicator of atopic dermatitis and psoriasis caused by the destruction of skin barrier function (see Non-Patent Document 2). LCE3D is also known as an indicator of psoriasis risk genes (see Non-Patent Document 3).

Interleukin-1 (Interleukin-1) is classified as a senescence-associated secretory phenotype (SASP). Specifically, it is known that interleukins 1a and 1b (IL-1a and IL-1b) as representative aging markers are involved in cellular senescence (see Non-Patent Document 4). It is also known that IL-1a, IL-1b, and PTGS2 are involved in skin inflammation, and XDH indicates oxidative stress induced by long-term exposure to a stimulus and is therefore external skin aging Indicators (see Non-Patent Document 5). Regarding skin inflammation, it is also known that the expression levels of PTGS2 (= COX-2) and IL-1a are increased in skin cells exposed to a stimulus (see Non-Patent Document 6). It is also known that XDH (xanthine dehydrogenase) as an indicator of oxidative stress is involved in skin aging.

List of cited materials Non-patent literature

Non-Patent Document 1: AM D’ Erme et al., “IL-36c (IL-1F9) Is a Biomarker for Psoriasis Skin Lesions”, Journal of Investigative Dermatology, Volume 135, 2015.

Non-Patent Document 2: Son et al., "S100A7 (psoriasin) inhibits human epidermal differentiation by enhanced IL-6 secretion through IjB / NF-jB signalling", Experimental Dermatology, John Wiley & Sons A / S, 2016.

Non-Patent Literature 3: Bergboer et al., "Psoriasis Risk Genes of the Late Cornified Envelope-3 Group Are Distinctly Expressed Compared with Genes of Other LCE Groups", The American Journal of Pathology, Vol. 178, No. 4, April 2011.

Non-Patent Document 4: Jean-Philippe Coppe et al., "The Senescence-Associated Secretory Phenotype: The Dark Side of Tumor Suppression", Annual Review of Pathology: Mechanisms of Disease, volume 5, 2010.

Non-Patent Literature 5: Kim, H.J. et al., "Transcriptome analysis of airborne PM2.5-induced detrimental effects on human keratinocytes", Toxicology Letters 273, 26-35, 2017.

Non-Patent Document 6: Natalia D Magnani et al., "Skin Damage Mechanisms Related to Airborne Particulate Matter Exposure", toxicological sciences, 149 (1), 2016, 227-236.

The inventors have found that fine dust is harmful to the skin, which in turn affects the expression of skin cell genes, causing symptoms such as skin cell damage.

Therefore, one aspect of the present invention is to provide a composition for caring for the damage of fine dust to skin cells.

In order to achieve the above object, one aspect of the present invention provides a composition for external use to care for the damage of fine dust to the skin, the composition containing wild mint extract as an active ingredient, and the composition is adjusted at least one selected from the following The performance of the formed group makes it reach the normal amount: IL-1a (NM_000575), IL-1B (NM_000576), IL-36G (NM_019618), S100A7 (NM_002963), LCE3D (NM_032563), PTGS2 (NM_000963), and XDH (NM_000379), which is a gene whose skin cells are affected by fine dust.

Hereinafter, the present invention will be described in detail.

Wild mint is a perennial herb of the Lamiaceae family of Tubiflorae. Also known as mint (mint or peppermint). Wild mint is characterized by a refreshing and cool taste. Its leaves and stems have a strong aroma when grown in a cool climate. It mainly contains menthol (menthol) and therefore emits a unique aroma. It also contains nutrients such as vitamins.

In one aspect of the present invention, the composition contains wild mint extract.

In one aspect of the present invention, leaves or stems of wild mint can be used, but not limited thereto.

Wild mint added with microorganisms can be specifically fermented at 22-30 ° C for 24-72 hours. In one embodiment, the extract is fermented at 26 ° C for about 48 hours.

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

In one aspect of the present invention, wild mint extract can be prepared by extracting wild mint leaves or stems with water or an organic solvent. Specifically, it can be prepared by extracting leaves or stems of wild mint with at least one extraction solvent selected from the group consisting of water, C ₁ -C ₆ anhydrous or hydrated lower alcohol, acetone, Butylene glycol, ethyl acetate, diethyl acetate, diethyl ether, benzene, chloroform, and hexane.

In one aspect, the wild mint extract can be extracted at room temperature.

In one aspect, the wild mint extract can be obtained by extraction with an extraction solvent and then further performing at least one of the following steps: filtration, concentration, separation, and drying. In particular, the wild mint extract can be subjected to at least one filtration step. In an embodiment, it undergoes two filtration steps.

In an embodiment, the separation step may include a centrifugation step.

Specifically, extraction may use at least one of the following polar solvents and low-polar solvents: polar solvents include water, C ₁ -C ₆ anhydrous or hydrated lower alcohols, acetone, and butylene glycol, and low-polar solvents include ethyl acetate , Diethyl acetate, ether, benzene, chloroform, and hexane.

More specifically, the solvent may be a 50-90% aqueous ethanol solution, and may be a 60-80% or 65-75% aqueous ethanol solution. When the solvent is 50-90% ethanol aqueous solution, the active ingredient can be effectively extracted from wild mint . In an embodiment, the solvent may be about 70% aqueous ethanol.

In one aspect, the extract can be concentrated under reduced pressure in a distillation apparatus equipped with a cooling condenser at an appropriate temperature after extraction.

However, the wild mint extract according to the present invention can be obtained by extraction by a method known in the art, and the extraction method is not limited to the above method.

In one aspect of the present invention, the composition may contain 0.00001 to 30% by weight of wild mint extract based on the total weight of the composition. When its content is 0.000001 to 30% by weight, wild mint extract has an excellent effect of caring for the damage of fine dust to the skin.

Specifically, the content may be 0.0000001 wt% or more, 0.0000005 wt% or more, 0.0000007 wt% or more, 0.0000009 wt% or more, 0.000001 wt% or more, 0.000002 wt% or more, 0.000004 wt% Percentage or more, 0.000006% by weight or more, 0.000008% by weight or more, 0.00001% by weight or more, 0.00003% by weight or more, 0.00005% by weight or more, 0.00007% by weight or more, 0.00009% by weight Or more, 0.0001% by weight or more, 0.0003% by weight or more, 0.0005% by weight or more, 0.0007% by weight or more, 0.0009% by weight or more, 0.001% by weight or more, 0.01% by weight or More, 0.1% by weight or more, 1% by weight or more, 3% by weight or more, 5% by weight or more, 7% by weight or more, 9% by weight or more, 10% by weight or more More, 13% by weight or more, 15% by weight or more 17% by weight or more, 19% by weight or more, 21% by weight or more, 23% by weight or more, 25% by weight or more, 27% by weight or more, 29% by weight or more, 30% by weight % By weight or more, or 31% by weight or more, and 32% by weight or less, 31% by weight or less, 30% by weight or less, 29% by weight or less, 28% by weight or less, 26% by weight or less, 24% by weight or less, 22% by weight or less, 20% by weight or less, 18% by weight or less, 16% by weight or less, 14% by weight or less, 12 Weight percent or less, 10 weight percent or less, 9 weight percent or less, 8 weight percent or less, 6 weight percent or less, 4 weight percent or less, 2 weight percent or less, 1 weight Percent or less, 0.1 percent by weight or less, 0.09 percent by weight or less, 0.04 percent by weight or less, 0.01 percent by weight or less, 0.006 percent by weight or less, 0.001% by weight or less, 0.0009% by weight or less, 0.0007% by weight or less, 0.00005% by weight or less, 0.00003% by weight or less, 0.00001% by weight or less, 0.000009% by weight or less, 0.000007 Weight percent or less, 0.000005 weight percent or less, 0.00003 weight percent or less, 0.000001 weight percent or less, 0.0000009 weight percent or less, 0.0000007 weight percent or less, 0.0000005 weight percent or less, 0.0000003 weight Percentage or less, 0.0000002 weight percent or less, 0.0000001 weight percent or less, 0.00000009 weight percent or less, but not limited thereto.

Another aspect of the present invention includes using the composition to care for the damage of fine dust to the skin.

Another aspect of the present invention provides a method for treating skin damage caused by fine dust on a subject, the method comprising administering an effective amount of wild mint extract to a subject in need thereof.

Another aspect of the present invention provides the use of wild mint extract in the manufacture of a composition, which is used to care for the damage of fine dust to the skin.

Another aspect of the present invention provides wild mint extract for the care of skin damage caused by fine dust.

As used in this article, the term "dust" refers to very small substances that are not visible to the naked eye. It is a particulate matter that floats or flutters in the air for a long time and has a diameter of 10 μm or less. In particular, particulate matter with a diameter of 2.5 μm or less is called “ultrafine particles”. In the present invention, "fine dust" is intended to include "ultrafine particles".

As used herein, the term "care" refers to effectively protecting skin cells from irritation, and inhibiting, preventing, or restoring (recovery) changes in specific gene expression due to stimulation.

In one aspect, the present invention provides a composition for suppressing damage to skin cells by fine dust, which is suppressed by regulating the expression level of specific genes in skin cells damaged by fine dust to a normal amount.

Specifically, in the present invention, those whose gene expression levels in skin cells are affected by dust include: IL-1a (NM_000575), IL-1B (NM_000576), IL-36G (NM_019618), S100A7 (NM_002963), LCE3D ( NM_032563), PTGS2 (NM_000963), and XDH (NM_000379). IL-1a (NM_000575), IL-1B (NM_000576), IL-36G (NM_019618), S100A7 (NM_002963), LCE3D (NM_032563), PTGS2 (NM_000963), and XDH (NM_000379) are genes whose performance increases due to dust . Therefore, it is possible to suppress the damage of skin cells by suppressing the expression of these genes to a normal amount.

Table 1 shows the genes used in the present invention and their expression levels increased due to fine dust. Table 1 shows the genes whose expression level increases due to fine dust. In this table, the term "name" refers to NCBI's gene bank login ID, the term "gene symbol" refers to the official gene symbol, and the term "gene title" refers to the name of each gene . These are described in Non-Patent Document 1. Table 1

Various analytical methods known in the art can be used to analyze the expression of genes or proteins, such as microarray, PCR, NGS (next generation sequencing), western blot, and northern blot (Northern blot), ELISA, radioimmunoassay, radioimmunodiffusion, immunohistochemical staining, immunoprecipitation assay, etc.

Dust can cause damage to skin cells, which can cause inflammation and further damage to skin cells. By taking care of the vicious cycle of skin cell damage with wild mint extract, skin conditions can be improved.

Another aspect of the invention includes the use of a composition to resist aging and inflammation.

Another aspect of the present invention provides an anti-aging method for a subject, the method comprising administering an effective amount of wild mint extract to a subject in need thereof.

Another aspect of the present invention provides the use of wild mint extract in the manufacture of a composition, the use being for anti-aging.

Another aspect of the present invention provides wild mint extract for anti-aging.

Another aspect of the present invention provides an anti-inflammatory method for a subject, the method comprising administering an effective amount of wild mint extract to a subject in need thereof.

Another aspect of the present invention provides the use of wild mint extract in the manufacture of a composition, the use being for anti-inflammatory.

Another aspect of the present invention provides wild mint extract for anti-inflammatory.

In one aspect, the present invention provides a composition that suppresses inflammation or aging by regulating the expression level of a specific gene in skin cells damaged by inflammation stimulation or aging to a normal amount.

In one aspect of the present invention, the expression levels of genes in skin cells affected by inflammation, stimulation or aging specifically include: IL-1a (NM_000575), IL-1B (NM_000576), PTGS2 (NM_000963), and XDH ( NM_000379). IL-1a (NM_000575), IL-1B (NM_000576), PTGS2 (NM_000963), and XDH (NM_000379) are genes whose expression is increased by inflammation or aging. Therefore, it is possible to suppress the inflammation or aging of skin cells by suppressing the expression levels of these genes to normal amounts.

In one aspect, the genes used in the present invention and their expression levels are increased by inflammation stimulation or aging are shown in Table 2. In this table, the term "name" indicates the NCBI gene bank registration ID, the term "gene symbol" indicates the official gene symbol, and the term "gene name" indicates the name of each gene. Table 2

Another aspect of the invention includes the use of the composition of the invention to strengthen skin barriers.

In one aspect, the present invention provides a composition for enhancing skin barriers by regulating the expression level of specific genes in skin cells damaged by weak skin barrier stimulation to a normal amount.

In one aspect of the present invention, the expression level of genes in skin cells affected by weak stimulation of induced skin barriers specifically includes: S100A7 (NM_002963), IL-36G (NM_019618) and LCE3D (NM_032563). S100A7 (NM_002963), IL-36G (NM_019618), and LCE3D (NM_032563) are genes whose expression is increased by weak stimulation of induced skin barriers. Therefore, skin barriers can be enhanced by suppressing the expression of these genes to normal amounts.

In one aspect, the genes used in the present invention are shown in Table 3 and their expression levels are increased by weak stimulation of induced skin barriers. In this table, the term "name" indicates the NCBI gene bank registration ID, the term "gene symbol" indicates the official gene symbol, and the term "gene name" indicates the name of each gene. table 3

Another aspect of the present invention provides a method for strengthening a skin barrier, the method comprising administering an effective amount of wild mint extract to a subject in need thereof.

Another aspect of the present invention provides the use of wild mint extract in the manufacture of a composition, which is used to strengthen skin barriers.

Another aspect of the present invention provides wild mint extract for strengthening skin barriers.

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

Examples of the cosmetic composition include cosmetics such as various creams, lotions, and toners, as well as cleaning products, cleanser, soap, and cosmetic solution.

In one aspect, the cosmetic containing the composition containing the wild mint extract of the present invention may be in the form of a solution, emulsion, viscous mixture, or the like.

That is, in one aspect, the cosmetic formulation of the present invention is not particularly limited and may be, for example, lotion, cream, toner, essence, pack, gel, powder (powder), makeup base (makeup base), foundation (foundation), lotion (lotion), ointment (ointment), patch (patch), makeup solution (cosmetic solution), cleansing foam (cleansing foam), Cleansing cream, cleansing water, body lotion, body cream, body oil, body essence, wash Shampoo, rinse, body cleanser, soap, hair dye or spray.

Depending on the desired formulation or purpose, those skilled in the art can select ingredients other than wild mint extract without difficulty and add them to the cosmetic composition of each formulation.

Furthermore, in one aspect, the cosmetic of the present invention may include a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polypeptides, polysaccharides, sphingolipids, and seaweed extracts.

Furthermore, in one aspect, if necessary, the cosmetic of the present invention may contain ingredients other than essential ingredients that are commonly used in cosmetics.

Examples of additional ingredients include oil ingredients, moisturizers, lubricants, surfactants, organic and inorganic pigments, organic powders, UV absorbers, preservatives, bactericides, antioxidants, plant extracts, pH adjusters, alcohols, Colorants, fragrances, blood circulation stimulants, skin coolers, antiperspirants, purified water, etc.

However, the ingredients that can be contained in cosmetics are not limited to this. Also, the amount of any of these ingredients can be determined within a range that does not negatively affect the purpose and effects of the present invention.

In one aspect, the pharmaceutical composition containing the wild mint extract of the present invention may further include a suitable carrier, excipient, and diluent conventionally used for preparing the pharmaceutical composition.

The pharmaceutical composition containing wild mint extract can be formulated into any form suitable for pharmaceutical preparations according to conventional methods, including oral formulations such as lozenges, capsules, powders, or syrups, and agents for external use on the skin such as Ointments, gels, creams, patches, and sprays.

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

In the health functional food composition according to the aspect of the present invention, the health food may refer to foods made from nutrients that may be lacking in a normal diet, or raw materials or ingredients (functional raw materials) having functions useful to the human body, and By maintaining the normal functions of the human body or activating physiological functions to maintain and improve health, but not limited to this. Health foods can be manufactured and processed into tablets, capsules, powders, granules, liquids, pills, etc. However, the formula is not limited to this, and can be manufactured and processed into any form under the law.

Specifically, the health drink composition is not particularly limited in composition except for the above-mentioned compound contained as an essential ingredient in a predetermined ratio. It may contain various flavoring agents or natural carbohydrates as additional ingredients in conventional beverages. Examples of natural carbohydrates are conventional sugars such as monosaccharides, polysaccharides, and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. Also, natural flavoring agents (thaumatin, stevia extracts (eg, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (eg, saccharin ), Aspartame, etc.) can be used as a flavoring agent.

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

Hereinafter, the composition and effects of the present invention will be described in more detail with reference to examples. However, the following examples are provided for illustrative purposes only to facilitate understanding of the present invention, and the scope of the present invention is not limited thereto. Example 1: Preparation of wild mint extract

Wild mint leaves and Acetobacter aceti were prepared and fermented at 26 ° C for 48 hours. Then, using an extraction solvent obtained by mixing purified water and ethanol (that is, 70% ethanol) at a ratio of 3: 7 as an extraction solvent, the fermentation product was extracted at room temperature. After extraction at room temperature, filtration is performed once to remove solid substances contained in the extract. Then, the extract is concentrated to remove ethanol, followed by separation and purification. Then, the resultant was subjected to centrifugation and secondary filtration, and then dried to obtain wild mint extract. Example 2: Fine dust collection and extraction

Use a low-volume air sampler (Sensidyne, Gillian, FL, U.S.A.) to collect fine dust. The filter and the filter of the filter pack were replaced at about 10:00 am on each measurement day to collect samples for about 24 hours. In the Shunfeng District of Seoul (International Research Center of the Korea University of Foreign Studies, the roof of the 6th floor of the Dormitory Building in Gyeonggi-do, Gyeonggi-do, Yongin City), dust was collected for 28 days every day. The timer is started when the vacuum pump is turned on and the timer time is recorded when the vacuum pump is turned off to measure the measurement time. The collection flow rate is set at 16.7 L / min. The flow rate was measured with a flow meter (model 4143, TSI Inc.) at the beginning of the measurement, and measured again at the end of the measurement. The Teflon filters in the filter set were weighed before and after collection. Dry the Teflon filter in a desiccator (NIKKO, Japan) at a relative humidity of 40% to a constant weight for 24 hours, then measure the weight of the Teflon filter, and then place it on a five-digit electronic balance (DVG215CD , Ohaus) weighed twice. Record the average. After collecting the sample, before measuring the weight, the Teflon filter was dried again in the dryer to a constant weight for 24 hours, and then weighed twice. Next, the mass concentration is calculated by comparing the measured weight with the weight measured before collection. The dust extraction is as follows: Wet the Teflon filter with 1 mL of ethanol and place 14 mL of DW so that the surface of the aerosol collector of the filter reaches the surface of the water, close the lid, and then use the ultrasonic cleaner to oscillate the filter 30 minute. In order to minimize errors caused by moisture in the filtration step, the moisture is completely removed from the filter in the dryer for 48 hours. Then, the weight of the filter is measured using an ultra-precision balance (Mettler Toledo Company) that can measure 0.1 mg to measure the weight of the filter before and after extraction. Example 3: Cultivation of keratinocyte line (normal person)

The keratinocyte cell line (normal human) was purchased from Lonza Inc. (Walkersville, Maryland, USA), subcultured, and then cultured in a CO ₂ incubator under the conditions of 37 ° C. and 5% CO ₂ . Follow Lonza's guidelines for cell culture. KGM-2 bullet kit CC-4152 (ingredient: BPE (bovine pituitary extract)) to which GA-1000 (gentamycin suflate + amphotericin-B (amphotericin-B)) will be added , Human epidermal growth factor (hEGF), insulin, hydrocortisone, transferrin, epinephrine, and KGM-2 bullet kit CC-3107, added to 500ml KBM-2 CC-3103 medium to prepare medium for cell culture. Example 4: Treating keratinocyte cell lines (normal people) with fine dust and measuring cytotoxicity

In order to study the cytotoxicity treated with fine dust, the MTT test was performed with a keratinocyte cell line (normal human) according to the method of Mossman et al. (J. Immunol. Methods, 65, 55-63, 1983).

Specifically, a 24-well disk is used. The fine dust having a diameter of 2.5 μm obtained in Example 2 was dispersed in purified water to prepare a fine dust dispersion liquid. Then, the fine dust dispersion liquid was applied to the cells cultured under the conditions of Example 3 and 2.5 × 10 ⁵ cells per well, followed by culturing for 24 hours. Then, the cells were mixed with 5 mg / ml of MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazole (3- (4,5-Dimethylthiazol-2 -yl) -2,5-diphenyltetrazolium bromide)) were mixed and further incubated at 37 ° C for 3 hours. The medium was then removed, and the formed formazan crystal was dissolved in 500 μl of DMSO. The lysate was aliquoted into 96-well dishes, and the OD value was measured at 540 nm. The measurement results are shown in Figure 1.

As shown in FIG. 1, for the cytotoxicity caused by the dispersion obtained by dispersing fine dust of 2.5 μm or less, the concentration (IC20) at which the cell line reached 80% survival rate was 12.5 μg / ml. Example 5: Study the changes of cell genes due to fine dust through the next generation sequencing method

For processing and analysis of RNA base sequence data, refer to the general analysis procedure developed by Trapnell et al. (2012). FastQC was used to determine the quality of RNA-seq data (http://www.bioinformatics.babraham.ac.uk/projects/fastqc/). Use FASTX to remove base and adaptor sequences with low accuracy (http://hannonlab.cshl.edu/fastx_toolkit/). Then, Tophat (Trapnell et al., 2009) and human genome (hg19) were used for comparison, and RSeQC, renamed EVER-seq, was used to determine the data volume of each sample (Wang et al., 2012). In addition, Cufflinks were used to quantify the expression level of transcripts, and to compare the transcription level between samples treated with fine dust dispersion and normal samples (Trapnell et al., 2010). Stringent cut-offs with FDR adjusted p-values <0.05 and ≥2.0 times-change were used to determine genes that showed significant expression differences when treated with a dispersion of fine dust with a diameter of 2.5 μm. The measurement results are shown in Table 4 below and FIGS. 2A to 2G. Table 4 Example 6: Real-time polymerase chain reaction (RT-PCR) quantification

The normal human keratinocytes cultured in Example 3 were treated with fine dust with a diameter of 2.5 μm extracted in Example 2 in an amount of 12.5 μg per 1 ml of cell culture medium. Then, the relative mRNA expression level was measured using primers (applied biosystems TaqMan® Primers) of the genes indicated in Table 5 below. The extract prepared in Example 1 was used as wild mint extract. table 5

Treat the medium with 20 ppm wild mint extract. After 24 hours, the culture medium was removed, and the cells were washed with 2 ml of phosphate buffered saline (PBS). Then, RNA was isolated from the cells using Trizol reagent (Invitrogen, Carlsbad, CA, USA). The isolated RNA was further purified using RNA kit (QIAGEN RNeasy kt, QIAGEN, Valencia, CA). Then, the quality of RNA was measured using Agilent 2100 BioAnalyzer (Agilent Technologies, Santa Clara, CA, USA). CDNA was synthesized from RNA using a reverse transcription kit (Superscript Reverse Transcriptase (RT) kit, Invitrogen, Carlsbad, CA). The primers shown in Table 5 were used for quantitative analysis of cDNA by real-time reverse transcription polymerase chain reaction (Q-RT-PCR). The TaqMan Gene Expression Detection Kit (Applied Biosystems, Foster City, CA) was used to assess changes in cell genes by real-time PCR. The results are shown in Figure 3. Both Q-RT-PCR and real-time PCR are performed according to the standard protocol issued by Life Technologies. Specifically, 40 following cycles were performed: 95 ° C for 20 seconds, 95 ° C for 3 seconds, and 60 ° C for 30 seconds.

Figures 3A to 3G show that there are genes whose expression level increases or decreases in skin cells stimulated by fine dust. In addition, interleukin-1α (IL-1a), interleukin 1β (IL-1B), interleukin 3β-36γ (IL-36G), S100 calcium binding protein A7 (S100A7), advanced keratin were found Envelop protein 3D (LCE3D), prostaglandin endoperoxide synthase 2 (PTGHS2), and xanthine dehydrogenase (XDH) gene expression levels were treated with wild mint extract And lower.

In addition, FIGS. 3A to 3G show that there is a gene whose expression level increases in stimulated skin cells that are stimulated by inflammation or aging. Also, interleukin-1α (IL-1a), interleukin-1β (IL-1B), prostaglandin endoperoxide synthase 2 (PTGHS2), and yellow The expression level of the purine dehydrogenase (XDH) gene is reduced by treatment with wild mint extract.

Therefore, it has been found that wild mint extract effectively protects skin cells from skin cell damage caused by inflammation or aging stimuli, and inhibits or prevents the expression level of the above-mentioned specific genes from being changed due to inflammation or aging stimuli to achieve normal performance .

In addition, FIGS. 3A to 3G show that there is a gene whose expression level increases in stimulated skin cells that are stimulated by weak skin barriers. Also, it was found that the expression levels of interleukin 36γ (IL-36G), S100 calcium binding protein A7 (S100A7), and late keratinocyte envelope protein 3D (LCE3D) genes were reduced by treatment with wild mint extract.

Therefore, it has been found that wild mint extract effectively protects skin cells from skin cell damage caused by weak skin stimulation induced by skin barriers, and inhibits or prevents the expression level of the above-mentioned specific genes from changing due to weak skin stimulation induced by skin barriers to achieve normal Performance.

Therefore, it was found that the extract of wild mint effectively protects skin cells from the stimulation of fine dust, and inhibits or prevents the expression level of the above-mentioned specific genes from being changed due to the stimulation, so as to achieve a normal expression level.

Hereinafter, formulation examples of the composition according to the present invention will be described. However, the cosmetic composition, the pharmaceutical composition, and the health care functional food composition can be formulated into various other forms. These examples are for illustrative purposes only, and are not intended to limit the scope of the present invention. Formulation example 1: lozenges

100 mg of wild mint extract according to an embodiment of the present invention, 400 mg of lactose, 400 mg of corn starch, and 2 mg of magnesium stearate are mixed, and subjected to a tabletting process according to the conventional method of preparing tablets ( tableting process) to prepare lozenges. Table 6 Formulation example 2: capsule

Mix 100 mg of wild mint extract according to an embodiment of the present invention, 400 mg of lactose, 400 mg of corn starch, and 2 mg of magnesium stearate, and fill it into gelatin capsules according to the conventional method of preparing capsules To prepare capsules. Table 7 Formulation example 3: granules

50 mg of wild mint extract according to an embodiment of the present invention, 250 mg of anhydrous crystalline glucose, and 550 mg of starch were mixed and formulated into granules using a fluidized bed granulator. The granules are then packed into sachets. Table 8

Formulation Example 4: Soap Table 9

Formulation Example 5: Emulsion Table 10

Formulation Example 6: Cream Table 11

Formulation Example 7: Ointment Table 12

Formulation Example 8: Preparation of cosmetic solution Table 13

Formulation Example 9: Health Food Table 14

Formulation Example 10: Health Drink Table 15

Although the present invention has been described in terms of specific embodiments, it is obvious to those having ordinary knowledge in the technical field that it can be carried out without departing from the spirit and scope of the present invention as defined in the following patent application scope Various changes and modifications.

no

Figure 1 shows cell viabilities when treated with microdust extracts. Here, ADSP (which stands for Asian dust storm particle) refers to Asian dust, PM10 (particulate matter 10) refers to fine dust with a particle size of 10 μm, and PM2. 5 (particulate matter 2.5) refers to fine dust with a particle size of 2.5 μm; Figure 2A shows the increase in mRNA expression of the S100A7 gene in skin cells stimulated by PM2.5 fine dust; Figure 2B shows in skin cells stimulated by PM2.5 fine dust The mRNA expression level of IL-1A gene increases; Figure 2C shows the increased mRNA expression level of IL-1B gene in skin cells stimulated by PM2.5 dust; Figure 2D shows the IL-36G gene expression in skin cells stimulated by PM2.5 dust MRNA expression level increased; Figure 2E shows an increase in the mRNA expression level of the PTGS2 gene in skin cells stimulated by PM2.5 dust; Figure 2F shows an increase in the mRNA expression level of the LCE3D gene in skin cells stimulated by PM2.5 dust; Figure 2G It shows that the expression level of XDH gene mRNA in skin cells stimulated by PM2.5 dust is increased; Figure 3 shows that by treatment with wild mint extract, the gene expression level changed in skin cells stimulated by dust is back to normal; Figure 3A shows By treatment with wild mint extract, S100A7 gene expression levels in skin cells stimulated by dust in the changed back to a normal amount; FIG. 3B shows, by treatment with wild mint extracts, skin cell stimulated by dust altered The IL-1A gene expression level returned to the normal amount; Figure 3C shows that by treatment with wild mint extract, the changed IL-1B gene expression level in the skin cells stimulated by fine dust returned to the normal amount; Figure 3D shows that By treatment with wild mint extract, the amount of IL-36G gene expression in skin cells stimulated by fine dust returned to normal; Figure 3E shows that, by treatment with wild mint extract, skin cells stimulated by fine dust The altered PTGS2 gene expression returns to the normal amount; Figure 3F shows that by treatment with wild mint extract, the altered LCE3D gene expression in the skin cells stimulated by fine dust returns to the normal amount; and Figure 3G shows that by After treatment with wild mint extract, the amount of XDH gene expression in skin cells stimulated by fine dust returned to normal.

Claims (19)

A composition for treating the damage of fine dust to skin cells, which contains wild mint extract as an active ingredient. The composition according to claim 1, wherein the damage of the care dust to skin cells is at least one selected from the group consisting of: skin barrier enhancement, anti-aging, and anti-inflammatory. A composition for strengthening the skin barrier, which contains wild mint extract as an active ingredient. An anti-aging composition comprising wild mint extract as an active ingredient. A composition for anti-inflammatory, which contains wild mint extract as an active ingredient. The composition according to any one of claims 1 to 5, wherein the content of the wild mint extract is 0.000001 to 30% by weight based on the total weight of the composition. The composition according to any one of claims 1 to 5, wherein the wild mint extract is extracted with at least one extraction solvent selected from the group consisting of water, C ₁ -C ₆ anhydrous or hydrated low grade Alcohol, acetone, butylene glycol, ethyl acetate, diethyl acetate, ether, benzene, chloroform, and hexane. The composition according to any one of claims 1 to 5, wherein the wild mint extract is extracted from the leaves of wild mint . The composition according to any one of claims 1 to 5, wherein the wild mint is fermented wild mint and fermented by adding a microorganism of the genus Acetobacter sp . The composition according to the requested item 9, wherein a microorganism of the genus acetate based Acetobacter (Acetobacter aceti). The composition according to any one of claims 1 to 2, wherein the composition inhibits at least one performance selected from the group consisting of: IL-1a (NM_000575), IL-1B (NM_000576), L -36G (NM_019618), S100A7 (NM_002963), LCE3D (NM_032563), PTGS2 (NM_000963), and XDH (NM_000379). The composition of claim 3, wherein the composition inhibits at least one performance selected from the group consisting of L-36G (NM_019618), S100A7 (NM_002963), and LCE3D (NM_032563). The composition according to claim 4, wherein the composition inhibits at least one performance selected from the group consisting of: IL-1a (NM_000575), IL-1B (NM_000576), PTGS2 (NM_000963), and XDH ( NM_000379). The composition according to claim 5, wherein the composition inhibits at least one performance selected from the group consisting of IL-1a (NM_000575), IL-1B (NM_000576), and PTGS2 (NM_000963). The composition according to any one of claims 11 to 14, wherein the composition acts on keratinocytes. The composition according to any one of claims 1 to 5, wherein the wild mint extract is administered at a dose of 10 to 500 mg / kg / day. The composition according to any one of claims 1 to 5, wherein the composition is a cosmetic composition. The composition according to any one of claims 1 to 5, wherein the composition is a pharmaceutical composition. The composition according to any one of claims 1 to 5, wherein the composition is a health-care functional food composition.