CN111315357A - Composition containing lactic acid bacteria derived from green tea for protecting skin cell damage caused by fine dust - Google Patents
Composition containing lactic acid bacteria derived from green tea for protecting skin cell damage caused by fine dust Download PDFInfo
- Publication number
- CN111315357A CN111315357A CN201880071530.1A CN201880071530A CN111315357A CN 111315357 A CN111315357 A CN 111315357A CN 201880071530 A CN201880071530 A CN 201880071530A CN 111315357 A CN111315357 A CN 111315357A
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- China
- Prior art keywords
- composition
- lactic acid
- acid bacteria
- green tea
- weight
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Abstract
Disclosed is a composition for protecting skin damage caused by mote, which contains, as an active ingredient, lactic acid bacteria derived from green tea, a lysate thereof, a culture thereof, or an extract thereof, and which is capable of regulating the expression level of a gene in skin cells affected by mote, the gene in skin cells affected by mote being selected from one or more genes of keratin 1(KRT1), claudin 1(CLDN1), claudin 4(CLDN4), and Occludin (OCL N), to a normal level. By using the composition for protecting skin damage caused by fine dusts according to the present invention, the gene expression level changed by fine dusts can be adjusted to a normal level, thereby caring for damaged skin cells.
Description
Technical Field
The invention discloses a composition for treating skin cell damage caused by dust particles. In particular, the present invention discloses a composition containing lactic acid bacteria derived from green tea, which can care for skin cell damage by significantly changing expression levels as biomarkers of skin cell genes, etc., compared to normal skin cells, due to dust, etc.
Background
Recently, due to environmental factors, concerns about fine dusts have been increasing, and contaminated areas have been expanding. The fine dusts have smaller sized particles than general yellow sands, etc., and thus can be more easily infiltrated into the human body, and are more harmful to the human body since they contain more harmful substances such as heavy metals, nitrates, ammonium, sulfates, etc. When the dust particles enter the body, the cells responsible for the immunity of the human body play a role in removing the dust particles. At this time, there are accompanying inflammatory side effects, and in addition, the world health organization classifies mottle as a group of carcinogens directly related to cancer.
Although it is expected that fine dusts penetrate not only into the respiratory tract but also into the skin or hair follicles to cause skin problems or hair loss, research on the mechanism and the target of their negative effects on the skin and hair is not complete, and thus, research on anti-pollution materials for solving the problems has not been conducted.
Lactic acid bacteria are bacteria that produce lactic acid (lactic acid) by decomposing sugars such as glucose, and are also called lactic acid bacteria. Lactic acid produced by lactic acid fermentation of lactic acid bacteria can inhibit the growth of pathogenic bacteria and harmful bacteria, and these characteristics are applied in the production of foods such as dairy products, kimchi, and brewed foods. In addition, since lactic acid bacteria prevent abnormal fermentation of bacteria by inhabiting the lining in the intestinal tract of mammals, they are also important bacteria used in intestinal conditioning agents.
Lactobacillus plantarum (Lactobacillus plantarum) is a strain of Lactobacillus, which is known to grow mainly when fermentation of kimchi proceeds to a large extent and sour taste appears. The optical isomers of lactic acid produced are the D form and the L form. Lactobacillus plantarum is known for its excellent acid resistance, bile acid resistance and antibacterial properties because it is widely used in various foods requiring fermentation, however, it is not known as an antipollution material having skin barrier enhancing effect and skin cell damage recovery effect, etc.
[ Prior art documents ]
[ non-patent literature ]
(non-patent document 1) Kim, H.J. et al, "Transcriptome analysis of harmful effects of atmospheric PM2.5 on human keratinocytes," toxicological Rapid report 273,26-35,2017 (Kim, H.J., et al, "transformed analysis of inorganic PM2.5-induced breakdown effects on human keratocytes", toxological letters 273,26-35,2017.)
Disclosure of Invention
Technical problem
Based on this, the present inventors have confirmed that while fine dust exerts a harmful influence on the skin and the expression of skin cell genes is also influenced by these influences, skin cell damage and the like are caused, lactic acid bacteria derived from green tea have an effect of restoring gene expression.
Accordingly, in one aspect, the present invention is directed to a composition for protecting skin cell damage caused by fine dusts, which contains lactic acid bacteria derived from green tea, a lysate thereof, a culture thereof, or an extract thereof.
Technical scheme
To achieve the above objects, in one aspect, the present invention provides a composition for protecting skin damage caused by mote, the composition comprising as an active ingredient a lactic acid bacterium derived from green tea, a lysate thereof, a culture thereof, or an extract thereof, the composition being capable of regulating the expression level of a gene in skin cells affected by mote to a normal level, the gene in skin cells affected by mote being one or more genes selected from the group consisting of keratin 1(KRT1), claudin 1(CLDN1), claudin 4(C LDN4), and occludin (ocln).
Advantageous effects
In one aspect, by using the composition for protecting skin damage caused by fine dusts provided by the present invention, the expression level of genes altered by fine dusts can be regulated to a normal level, thereby caring for damaged skin cells.
Drawings
Fig. 1a shows that the mRNA expression level of the cl audin 1(CLDN1) gene in skin cells was increased by treatment with a standard strain (type strain) or lactic acid bacteria derived from green tea.
Fig. 1b shows that the mRNA expression level of the cla udin 4(CLDN4) gene in skin cells was increased by treatment with standard strains (type strains) or lactic acid bacteria derived from green tea.
FIG. 1c shows that the mRNA expression level of Oc Claudin (OCLN) gene in skin cells was increased by treatment with a standard strain (type strain) or lactic acid bacteria derived from green tea.
Fig. 1d shows that the mRNA expression level of keratin 1(KRT1) gene in skin cells was increased by treatment with standard strains (type strains) or lactic acid bacteria derived from green tea.
FIG. 2a shows that the amount of mR NA expression of claudin 1(CLDN1) gene is reduced in skin cells stimulated by ERM-CZ120 mote, followed by restoration to normal levels by lactic acid bacteria treatment derived from green tea.
FIG. 2b shows that the amount of mR NA expression of claudin 4(CLDN4) gene was reduced in skin cells stimulated by ERM-CZ120 mote, and then returned to normal levels by lactic acid bacteria treatment derived from green tea.
FIG. 2c shows that mRNA expression level of Occludin (OCLN) gene was decreased in skin cells stimulated by ERM-CZ120 mote, and then returned to normal level by lactic acid bacteria treatment derived from green tea.
FIG. 2d shows that the mRNA expression level of keratin (KRT1) gene was decreased in the skin cells stimulated by ERM-CZ120 mote, and then returned to normal level by lactic acid bacteria treatment derived from green tea.
Detailed Description
Hereinafter, the present invention will be described in detail.
In one aspect of the present invention, the composition for caring for skin damage caused by the fine dusts may comprise, as an effective ingredient, a lactic acid bacterium derived from green tea, a lysate thereof, a culture thereof, or an extract thereof.
In the present specification, "lactic acid bacteria derived from green tea" may mean lactic acid bacteria isolated from tea leaves, tea flowers, tea tree trunks, tea tree roots, or soil in the vicinity, and the like. For example, it may refer to lactic acid bacteria cultured and isolated from green tea as a leaf of tea tree.
In the present specification, "strain lysate" may refer to a product obtained by lysing the strain itself using chemical or physical force.
In the present specification, "culture" may refer to a substance containing all substances contained in a medium in which a strain is cultured, for example, may refer to a substance containing metabolites or secretions that are culture products of the strain, or a lysate thereof, and the strain itself may also be included in the culture.
In the present specification, the term "extract" may refer to a product obtained by extracting the strain itself, a lysate of the strain, a culture thereof, or a mixture thereof with an extraction solvent.
The green tea lactic acid bacteria according to an embodiment of the present invention are lactic acid bacteria separated from various components contained in the above-mentioned leaves of tea tree, and when they are applied to the skin damaged by fine dust, particularly to the tight junction (light junctio n) of the skin barrier, their recovery efficacy for the damaged skin is extremely excellent.
In one aspect, the lactic acid bacteria derived from green tea may be Lactobacillus sp. Specifically, the lactic acid bacterium derived from green tea may be Lactobacillus plantarum (Lactobacillus plantarum). More specifically, the lactic acid bacterium derived from green tea may be lactobacillus plantarum APsulloc 331266(lactobacillus plantarum APsulloc 331266) KCC M111 11181P. The green tea-derived lactic acid bacteria can enhance tight junctions (light junctions) of the skin barrier, and especially lactobacillus plantarum APsulloc 3312666(lactobacillus plantarum APsulloc 331266) KCCM11181P can have the effect of restoring the skin barrier damaged by fine dusts.
In one aspect, the active ingredient of the composition may be lactic acid bacteria apoptotic bodies derived from green tea.
In one aspect of the present invention, in the composition, the composition may comprise 0.000001% to 30% by weight of the lactic acid bacteria derived from green tea, a lysate thereof, a culture thereof, or an extract thereof, based on the total weight of the composition. When the content is 0.000001 to 30% by weight, the care effect of the lactic acid bacteria derived from green tea on skin damage caused by fine dust is excellent.
Specifically, it may be 0.0000001% by weight or more, 0.0000005% by weight or more, 0.0000007% by weight or more, 0.0000009% by weight or more, 0.000001% by weight or more, 0.000002% by weight or more, 0.000004% by weight 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, or 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 or more, 31% by weight or more, and 2% 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% by weight or less, 10% by weight or less, 9% by weight or less, 8% by weight or less, 6% by weight or less, 4% by weight or less, 2% by weight or less, 1% by weight or less, 0.1% by weight or less, 0.09% by weight or less, 0.04% by weight or less, 0.01% by weight or less, 0.006% by weight or less, 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% or less, 0.00001 wt% or less, 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, 0.0000003 wt% or less, 0.0000002 wt% or less, 0.0000001 wt% or less, 0.00000009 wt% or less, but not limited thereto.
In one aspect, the active ingredient of the composition according to the invention may act on the tight junctions of the skin barrier.
Tight junctions are one of the cell junctions, consisting of occluding, claudin, tricellulin, Junctional Adhesive Molecules (JAM), and Zona Occludins (ZO). Tight junctions can be divided into cell membrane-containing and intracellular portions. occludin and claudin are tight junction constitutive proteins contained in cell membranes. occludin is the first transmembrane protein found in tight junctions, with four transmembrane domains. occludin is not only a component of tight junction, but also has the function of regulating tight junction. The cl audin, which is a tight junction component protein with occludin, also has four transmembrane domains. The tight junctions, which serve as connecting ligaments between cells, have the functions of filling gaps between adjacent cells, regulating the movement of small substances, and also play a role in regulating the permeation of substances between cells and maintaining the polarity of cells. Recently, according to the report of the research results, the tight junction plays an important role in the skin barrier function. Tight junctions are one of the modes of intercellular binding. Is a cellular junction that occurs primarily in epithelial and endothelial cells, forming a complex network with linear structures. Due to this structure, the tight connection has mainly two functions. The first is a barrier function, which prevents leakage of substances between cells constituting epithelial cells or endothelial cells. It is mainly treated in the blood vessels and digestive organs. The second is an occlusion function (function) which is a function of separating cell membranes by wrapping a linear rail in the lipids of the cell membranes. Therefore, the active ingredient of the composition according to the present invention enhances the barrier function and the blocking function of skin cells by acting on the tight junctions of the skin barrier, thereby protecting skin damaged by the stimulation of fine dusts.
In another aspect of the invention, the invention includes the use of the composition for the care of skin damage caused by mote.
In another aspect of the present invention, the present invention provides a method for caring for skin damage caused by mote in a subject, the method comprising the step of administering an effective amount of lactic acid bacteria derived from green tea, a lysate thereof, a culture thereof, or an extract thereof to a subject in need thereof.
In another aspect of the present invention, there is provided a use of a lactic acid bacterium derived from green tea, a lysate thereof, a culture thereof, or an extract thereof for the preparation of a composition for the protection of skin damage caused by fine dusts.
In another aspect of the present invention, there is provided a lactic acid bacterium derived from green tea, a lysate thereof, a culture thereof, or an extract thereof for use in the protection of skin damage caused by fine dusts.
As used herein, "dust" refers to an extremely fine substance that is invisible to our eyes and is a particulate substance that floats or flies in the atmosphere for a long period of time. Specifically, in the present invention, the particle diameter of the fine dust may be 10 μm or less (PM10), more specifically, 2.5 μm or less (PM 2.5). In particular, particulate matter having a particle size of 2.5 μm (PM2.5) or less is referred to as "ultrafine dust," and "fine dust" in the present specification also includes "ultrafine dust.
In one aspect of the invention, the dust particles may comprise one or more of arsenic (arsenic), cadmium (cadmium), lead (lead), and nickel (nickel). Since the fine dusts contain heavy metal components such as arsenic, cadmium, lead, and nickel, they attack skin cells to cause skin allergy, pigmentation, inflammation, and the like, thereby causing skin aging. In one embodiment, the mote comprises arsenic, cadmium, lead, and nickel.
The term "care" as used herein means to effectively protect skin cells from stimulation and to inhibit, prevent and restore (restore) the change in the expression level of a specific gene caused by the stimulation. The term "recovery" as used herein, with reference to the expression level of a gene without stimulation, includes not only changing the expression level closer to the above-mentioned standard, but also increasing the expression level to the above-mentioned standard or higher.
In one aspect, the present invention provides a composition for inhibiting skin cell damage caused by mote by regulating the expression level of a specific gene in skin cells damaged by mote to a normal level.
Specifically, in the present invention, in skin cells, a gene whose expression amount is affected by the mote is selected from one or more genes of keratin 1(KRT1), claudin 1(CLDN1), claudin 4(CLDN4) and Occludin (OCLN). since keratin 1(KRT1), claudin 1(CLDN1), claudin 4(CLDN4) and Occludin (OCLN) are genes whose expression amount is decreased by the mote, the expression amount can be adjusted to a normal level by increasing the expression amount of these genes, thereby inhibiting damage to skin cells.
In one aspect, the composition according to the present invention may have an efficacy of enhancing skin barrier or an efficacy of restoring skin barrier.
The following table [ Table 1] shows the genes whose expression levels were changed by the dust particles used in the present invention. In these tables, "Gene Symbol (Gene Symbol)" represents an official Gene Symbol and "Gene name (Gene title)" represents the name of each Gene. These contents can be confirmed as described in non-patent document 1.
[ TABLE 1]
Analysis of the expression amount of the gene or protein can be performed using various analysis methods known in the art, such as microarray, PCR, NGS (next generation sequencing), western blot, Northern blot hybridization, ELISA, radioimmunoassay, radioimmunodiffusion, immunohistochemical staining (IHC), immunoprecipitation analysis, and the like.
The fine dust may cause damage to skin cells and may cause inflammation at this time, thereby further causing damage to the skin cells. The malignant circulation of this skin cell injury can be treated by using lactic acid bacteria derived from green tea, thereby improving the skin condition.
In one aspect of the invention, the composition may be a cosmetic composition, a pharmaceutical composition or a functional health food composition.
The cosmetic composition includes, for example, various cosmetics such as cream, lotion (lotion), toner, and the like, and detergents, facial cleansers, soaps, and beauty lotions, and the like.
In one aspect, the cosmetic composition to which the composition containing the lactic acid bacteria derived from green tea according to the present invention is added may be used in the form of a solution, an emulsion, a viscous mixture, or the like.
That is, in one aspect, the cosmetic of the present invention is not particularly limited in its formulation, and may be, for example, in the form of emulsion, cream, lotion, essence, pack, gel, powder, pre-makeup emulsion, foundation, skin lotion, ointment, patch, beauty lotion, cleansing foam, makeup remover, body lotion, body cream, body oil, body essence, shampoo, hair conditioner, body wash, soap, hair dye, spray, and the like.
In the cosmetic compositions of various formulations, in addition to the lactic acid bacteria derived from green tea, those skilled in the art can suitably select other ingredients to formulate according to other cosmetic formulations or use purposes without difficulty.
Further, in one aspect, the cosmetic of the present invention may comprise a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high molecular peptides, high molecular polysaccharides, sphingolipids and seaweed extracts.
In one aspect, in the cosmetic according to the present invention, other ingredients generally used in cosmetics may be formulated as needed together with the above-described essential ingredients.
In addition, the compounding ingredients that can be added include organic ingredients, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, bactericides, antioxidants, plant extracts, pH adjusters, ethanol, pigments, perfumes, blood circulation promoters, coolants, antiperspirants, purified water, and the like.
Further, compounding ingredients that can be additionally added are not limited thereto, and any of the above-described ingredients may be compounded within a range that does not impair the object and effect of the present invention.
In one aspect, the pharmaceutical composition containing the lactic acid bacteria derived from green tea according to the present invention may further comprise suitable carriers, excipients and diluents commonly used for the preparation of pharmaceutical compositions.
As the formulation thereof, the pharmaceutical composition containing the lactic acid bacteria derived from green tea may be formulated into any form suitable for pharmaceutical preparations, such as oral preparations, such as tablets, capsules, powders, or syrups, or external preparations, such as ointments, gels, creams, patches, or sprays, according to a conventional method.
It will be understood that, when administered in the form of the pharmaceutical composition, the actual dose of the effective ingredient will generally be determined in accordance with various relevant factors such as the severity of symptoms, the chosen route of administration, the age, sex, body weight and health of the subject, etc. In general, the dosage of 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 functional health food composition according to an aspect of the present invention, the health food is a health food prepared using nutrients which are easily deficient in daily diet, or raw materials or ingredients (functional raw materials) having functions useful to the human body, and is a food capable of maintaining and improving health by maintaining normal functions or activating physiological functions of the human body, but not limited thereto. The health food may be prepared and processed into dosage forms of tablets, capsules, powders, granules, liquids, pills, etc., and is not limited thereto, and may be prepared and processed into any dosage form according to law.
Specifically, the health drink composition is not particularly limited to other ingredients except the compounds as essential ingredients in the specified ratios, and may contain various additional ingredients such as flavoring agents or natural carbohydrates as in general drinks. Examples of the natural carbohydrate include monosaccharides, disaccharides (e.g., glucose, fructose, etc.), polysaccharides (e.g., maltose, sucrose, etc.), common saccharides (e.g., dextrin, cyclodextrin, etc.), sugar alcohols (e.g., xylitol, sorbitol, erythritol, etc.), and the like. In addition to the above, the flavoring agent may use natural flavoring agents (thaumatin, stevia extract (e.g., rebaudioside a, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.).
It is to be understood that, when administered in the form of the functional health food composition, the actual dose of the effective ingredient should be generally determined according to various relevant factors such as the severity of symptoms, the chosen administration route, the age, sex, body weight and health condition of the subject, etc. 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.
Examples
Hereinafter, the constitution and effect of the present invention will be described in more detail by specific examples. However, these examples are only for illustrating the present invention, and the scope of the present invention are not limited by these examples.
[ PREPARATION EXAMPLE 1] Lactobacillus plantarum (Lactobacillus plantarum) APsulloc 331266
1. Culture and isolation of Lactobacillus plantarum APsulloc 331266
200 g of tea leaves were washed twice with primary distilled water to remove impurities. The water was removed from the washed tea leaves, and the tea leaves were mixed with 8% by weight of salt, and then left at room temperature for 3 hours. The salt-impregnated tea leaves were mixed with 1% of 1000mL fructo-oligosaccharide solution and cultured in an incubator at 32 ℃ for 3 days. After 3 days, it was confirmed whether the pH of the culture broth was less than 5, and when the pH was less than 5, it was taken out and added to Lactobacillus MRS agar of Difco
(Difco Lactobacilli MRS
) Culturing is carried out in a culture medium. At this time, the cells were cultured in the chamber at 32 ℃ under anaerobic conditions for 2 days, and then white colonies were removed.
Lactobacillus plantarum APsulloc 331266 was isolated from tea leaves in the same manner as described above. The lactobacillus plantarum AP sulloc 331266 was deposited at 2011 at 28.3.4 in Korean culture Collection No. KCCM 11181P.
2. Identification of Lactobacillus plantarum APsulloc 331266
(1) Cultivation of the Strain
APsulloc 331266 isolated in example 1 was streaked (streaking) on MRS agar plates and incubated at 37 ℃ for 2 days. The obtained single colony (single colony) was inoculated into 10mL of MRS broth (broth) and cultured again overnight at 37 ℃ to prepare an APsulloc 331266 culture solution.
(2) Sugar fermentation Pattern analysis of APsulloc 331266
The culture solution of APsulloc 331266 strain prepared as in (1) above was inoculated to 0.5% in 10mL of MRS broth and cultured overnight at 37 ℃. After the culture solution was centrifuged at 8,000rpm for 5 minutes, the supernatant was removed to collect only the cells, and then 2mL of 0.85% saline buffer (saline buffer) was added to suspend the cells. Thereafter, the experiment was performed using API50CHL kit (Biomerieux corporation) and according to the manufacturer's instructions. The specific contents thereof are as follows.
First, a suspension of the strain was slowly added to 5mL of API suspension medium (medium) while measuring the amount of the suspension required to reach the turbidity of McFarland Standard 2 (Biomerieux). The suspension was added twice as much as measured to 10mL of API50CHL medium and then mixed with shaking. After the above mixture was added to a cup (cup) containing different substrates, respectively, mineral oil was added dropwise and incubated at 37 ℃ for 2 days, after which the sugar fermentation pattern was analyzed.
The following table shows the results of comparing sugar fermentation patterns with APsulloc 331266 using lactobacillus plantarum strain (KCTC3108) as a standard strain, and APsulloc 331266 identified using the results.
[ TABLE 2 ]
+: decomposing a substrate; -: the corresponding substrate is not decomposed; is there a : can not determine
[ TABLE 3 ]
In addition, APSULLOC 331266 and the standard strain (KCTC3108) have different use characteristics for α -methyl-mannoside (mannose) and raffinose (raffinose), and thus it can be confirmed that the two strains are different.
3. Preparation of Lactobacillus plantarum APsulloc 331266 apoptotic bodies
The isolated and identified Lactobacillus plantarum APsulloc 33126 was cultured in medium (MRS broth) at 37 ℃ for 24 hours. The lactic acid bacteria to be inoculated were those stored in a freeze tube (Cryotube) containing skim milk at 10 wt%. When the concentration of bacteria in the liquid medium reached 5X108To 1X109Vegetative cells were collected by centrifugation at 4000rpm at 4 ℃ in CFU/mL, and the cells were washed with PBS (phosphate buffered saline). Then, the washed cells were treated at a ratio of 5X108To 1X109The concentration of CFU/mL was re-dispersed (re-suspended) in PBS and heated at 100 ℃ for 10 minutes to prepare Lactobacillus plantarum apoptotic bodies.
[ Experimental example 1] experiment for quantitative determination of Gene expression amount by real-time qPCR
1. Culture of keratinocyte cell lines
Keratinocytes (human normal epidermal keratinocytes) from Longsha corporation (Lonza, Inc., Wolville, Maryland, USA) were subcultured at 37 ℃ with 5% CO2Under the conditions of (1) CO2Culturing is carried out in a culture medium. A500 ml culture medium containing BPE (bovine pituitary extract), human epidermal growth factor (hEGF), Insulin (Insulin), Hydrocortisone (Hydrocortisone), gentamicin sulfate (Gentamycin sulfate), Epinephrine (Epinephrine), and Transferrin (Transferrin) was used in KBM-2(KBMTM-2, CC-3103) medium.
In the culture, the fusion degree (confluency) was maintained at about 90% at the maximum, and then subculture (subculture) was performed.
2. Treatment of fine dust and Lactobacillus plantarum in keratinocytes
The dust was ERM-CZ120 dust sold by Sigma Aldrich, which contained P M10 standard dust, arsenic, cadmium, lead and nickel.
Using 6-well plates, the cells were cultured at 3X 10 under the cell culture conditions of "1. culture of keratinocyte cell line5The conditioned cell strains of each cell/well (cell s/well) were divided into six groups and treated with substances, respectively, as an untreated group (control 1), a group treated with 50ppm dust (control 2), a group treated with 1. mu.g/ml of Lactobacillus plantarum of preparation example 1 (example 1), a group treated with 50ppm dust and 1. mu.g/ml of Lactobacillus plantarum of preparation example 1 (example 2), a group treated with 1. mu.g/ml Lactobacillus plantarum standard strain (type strain) (comparative example 1), and a group treated with 50ppm dust and 1. mu.g/ml Lactobacillus plantarum standard strain (comparative example 2).
Among them, the standard strain of Lactobacillus plantarum is a deposited strain obtained from the center for biological resources and having the deposit number KCTC 3108.
[ TABLE 4 ]
3. Real-time qPCR quantification
RNA extraction Process
The culture solution in control 1, control 2, example 1 and comparative example 1 obtained in "2. treatment of fine dusts and lactobacillus plantarum in keratinocytes" was removed and the cells were washed with 2ml of Phosphate Buffered Saline (PBS), and then RNA was isolated from the cells using Trizol reagent (Invitrogen, inc., calsbarded, california, usa).
3-2.cDNA Synthesis Process
The isolated RNA was purified again using an RNA kit from QIAGEN (QIAGEN RNeasy kt, QIAGEN, Valencia, Calif.) and then the quality of the RNA was confirmed using a model 2100BioAnalyzer (Agilent2100BioAnalyzer, Agilent Technologies, Santa Clara, Calif., USA). Thereafter, cDNA was synthesized from RNA using a reverse transcription kit (Superscript Reverse Transcriptase (RT) kit, zeimer fly (invitrogen), carlsbad, california, usa).
3-3. performing qPCR
Quantitative analysis was performed by real-time reverse transcription polymerase chain reaction (Q-RT-PCR) using primers. The change in gene expression pattern of the cells was evaluated by real-time PCR using TaqMan gene expression System (TaqMan gene expression assay kit) of Applied Biosystems, Foster City, Calif., and the results are shown in FIGS. 1a to 2 c. Both Q-RT-PCR and real-time PCR were performed according to the standard protocol published by Life Technino logies, and specifically, 40 cycles were performed by treating at 95 ℃ for 20 seconds, at 95 ℃ for 3 seconds, and at 60 ℃ for 30 seconds.
4-1 evaluation of skin cell barrier enhancing efficacy of Lactobacillus plantarum
As shown in fig. 1a to 1d, it was confirmed that both the standard strain and the lactic acid bacterium derived from green tea according to the present invention had the effect of enhancing the skin barrier when treated alone.
Specifically, the levels of mRNA expression of claudin1, claudin 4, ocludin and Keratin 1(Keratin 1) were increased in example 1 and comparative example 1 as compared with control 1.
4-2 evaluation of skin cell injury repair efficacy of Lactobacillus plantarum
As shown in fig. 2a to 2d, it was confirmed that the lactic acid bacteria derived from green tea according to the present invention have an effect of repairing skin barrier damaged by fine dust when treated alone.
Specifically, the mRNA expression levels of Keratin 1(Keratin 1), claudin1, claudin 4 and ocludin were confirmed to be increased in example 2 and comparative example 2 as compared with control group 2. In the experiment, it was confirmed that the mRNA expression level of keratin 1 was reduced to 0.41, the mRNA expression level of claudin1 was reduced to 0.57, the mRNA expression level of claudin 4 was reduced to 0.50, and the mRNA expression level of Occludin was reduced to 0.42 after the treatment with the fine dust, whereas the reduced mRNA expression level of keratin 1 was restored to 1.01, the reduced mRNA expression level of claudin1 was restored to 0.73, the reduced mRNA expression level of claudin 4 was restored to 1.18, and the reduced mRNA expression level of Occludin was restored to 0.79 after the treatment with lactobacillus plantarum in preparation example 1.
On the contrary, when treated with the standard strain, the amount of mRNA for keratin 1 was 0.51, the amount of mRNA for claudin1 was 0.53, the amount of mRNA for claudin 4 was 0.60, and the amount of mRNA for ocludin was 0.51, and the change in the amount of RNA expression was not significant. Thus, it was confirmed that the standard strain did not have the effect of restoring the reduced mRNA expression amount of the marker.
Dosage form examples
Examples of dosage forms of the composition according to the present invention are described below, and various dosage forms for cosmetic compositions, pharmaceutical compositions and health food compositions can also be applied, which are intended to describe the present invention in detail, but not to limit the present invention.
Dosage form example 1 tablets
100mg of Lactobacillus plantarum lactic acid bacteria according to an example of the present invention, 400mg of lactose, 400mg of corn starch and 2mg of magnesium stearate were mixed and then tabletted according to a conventional tablet preparation method.
[ TABLE 5 ]
| Compounding ingredients | Content (mg) |
| Lactobacillus plantarum lactic acid bacteria | 100 |
| Lactose | 400 |
| Corn starch | 400 |
| |
2 |
[ dosage form example 2 ] capsules
After 100mg of lactobacillus plantarum lactic acid bacteria according to an example of the present invention, 400mg of lactose, 400mg of corn starch, and 2mg of magnesium stearate were mixed, the mixture was filled in capsules according to a conventional capsule preparation method, and capsules were prepared.
[ TABLE 6 ]
| Compounding ingredients | Content (mg) |
| Lactobacillus plantarum lactic acid bacteria | 100 |
| Lactose | 400 |
| Corn starch | 400 |
| |
2 |
[ dosage form example 3 ] granules
50mg of Lactobacillus plantarum lactic acid bacteria according to an example of the present invention, 250mg of anhydrous crystalline glucose and 550mg of starch were mixed and prepared into granules using a fluidized bed granulator, and then filled into a coating to prepare granules.
[ TABLE 7 ]
| Compounding ingredients | Content (mg) |
| Lactobacillus plantarum lactic acid bacteria | 50 |
| Anhydrous crystalline glucose | 250 |
| Starch | 550 |
Formulation example 4 soap
[ TABLE 8 ]
| Compounding ingredients | Content (%) |
| Lactobacillus plantarum lactic acid bacteria | 5.00 |
| Oil and fat | Proper amount of |
| Sodium hydroxide | Proper amount of |
| Sodium chloride | Proper amount of |
| Perfume | Proper amount of |
| Purified water | Balance of |
Dosage form example 5 emulsion (deposition)
[ TABLE 9 ]
Dosage form example 6 cream
[ TABLE 10 ]
| Compounding ingredients | Content (%) |
| Lactobacillus plantarum lactic acid bacteria | 3.00 |
| Polyethylene glycol monostearate | 2.00 |
| Self-emulsifying glyceryl monostearate | 5.00 |
| Cetyl alcohol | 4.00 |
| Squalene | 6.00 |
| Triisooctanoic acid glyceride | 6.00 |
| Sphingolipid glycolipid | 1.00 |
| 1, 3-butanediol | 7.00 |
| Purified water | Balance of |
Dosage form example 7 ointments
[ TABLE 11 ]
| Compounding ingredients | Content (%) |
| Lactobacillus plantarum lactic acid bacteria | 5.00 |
| Polyvinyl alcohol | 13.00 |
| L-ascorbic acid-2-magnesium phosphate | 1.00 |
| Lauroyl hydroxyproline | 1.00 |
| Water-soluble collagen (1% water solution) | 2.00 |
| 1, 3-butanediol | 3.00 |
| Ethanol | 5.00 |
| Purified water | Balance of |
[ dosage form example 8 ] preparation of cosmetic liquid
[ TABLE 12 ]
| Compounding ingredients | Content (%) |
| Lactobacillus plantarum lactic acid bacteria | 3.00 |
| Hydroxyethylcellulose (2% aqueous solution) | 12.00 |
| Xanthan gum (2% aqueous solution) | 2.00 |
| 1, 3-butanediol | 6.00 |
| Concentrated glycerin | 4.00 |
| Sodium hyaluronate (1% aqueous solution) | 2.00 |
| Purified water | Balance of |
[ dosage forms example 9 ] health foods
[ TABLE 13 ]
| Compounding ingredients | Content (wt.) |
| Lactobacillus plantarum lactic acid bacteria | 2mg |
| Vitamin A acetate | 70μg |
| Vitamin E | 1.0mg |
| Vitamin B1 | 0.13 |
| Vitamin | |
| 2 | 0.15mg |
| Vitamin B6 | 0.5mg |
| Vitamin 12 | 0.2μg |
| Vitamin C | 10mg |
| Biotin | 10μg |
| Nicotinamide | 1.7mg |
| Folic acid | 50μg |
| Calcium pantothenate | 0.5mg |
| Ferrous sulfate | 1.75mg |
| Zinc oxide | 0.82mg |
| Magnesium carbonate | 25.3mg |
| Potassium dihydrogen phosphate | 15mg |
| Calcium hydrogen phosphate | 55mg |
| Potassium citrate | 90mg |
| Calcium carbonate | 100mg |
| Magnesium chloride | 24.8mg |
[ dosage form example 10 ] health drink
[ TABLE 14 ]
| Compounding ingredients | Content (wt.) |
| Lactobacillus plantarum lactic acid bacteria | 50mg |
| Citric acid | 1000mg |
| Fructo-oligosaccharide | 100g |
| Taurine | 1g |
| Purified water | Balance of |
[ accession number ]
The name of the depository institution: korea center for preservation of microorganisms (foreign)
The preservation number is: KCCM11181P
Storage date 20110328
[ microorganism preservation identification ]
Claims (14)
1. A composition for caring skin injury caused by mote comprises lactic acid bacteria derived from green tea, its lysate, its culture or its extract as effective component.
2. The composition according to claim 1, wherein the lactic acid bacteria derived from green tea are lactic acid bacteria of the genus Lactobacillus.
3. The composition according to claim 2, wherein the lactobacillus is a lactobacillus plantarum.
4. The composition according to claim 1, wherein the lactic acid bacteria derived from green tea is Lactobacillus plantarum APsullo c 331266 KCCM 11181P.
5. The composition according to claim 1, wherein the active ingredient is an apoptotic body of lactic acid bacteria derived from green tea.
6. The composition according to claim 1, wherein the lactic acid bacteria derived from green tea, its lysate, its culture or its extract is contained in an amount of 0.000001 to 30 wt% based on the total weight of the composition.
7. The composition according to claim 1, wherein the active principle of the composition acts on the tight junctions of the skin barrier.
8. The composition according to claim 1, wherein the composition is for restoring expression of one or more of keratin 1, claudin1, claudin 4 and Occludin.
9. The composition of claim 1, wherein the mote comprises one or more of arsenic, cadmium, lead, and nickel.
10. The composition of claim 1, wherein the particulate size of the particulate is PM10 or less.
11. The composition according to claim 1, wherein the active principle of the composition is administered in a dose ranging from 10 to 500 mg/kg/day.
12. The composition of claim 1, wherein the composition is a cosmetic composition.
13. The composition of claim 1, wherein the composition is a pharmaceutical composition.
14. The composition of claim 1, wherein the composition is a functional health food composition.
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| KR1020170143715A KR101993270B1 (en) | 2017-10-31 | 2017-10-31 | Composition comprising lactic acid bacteria derived from green tea for caring damages of skin cells by microdust |
| KR10-2017-0143715 | 2017-10-31 | ||
| PCT/KR2018/009465 WO2019088421A1 (en) | 2017-10-31 | 2018-08-17 | Composition for treating skin cell damage caused by fine dust, containing green tea-derived lactic acid bacteria |
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| KR20210104953A (en) | 2020-02-17 | 2021-08-26 | (주)아모레퍼시픽 | Composition for relieving itching or inflammation caused by microdust |
| KR102668566B1 (en) * | 2022-07-06 | 2024-06-10 | 주식회사 현대바이오랜드 | Lactobacillus plantarum HDB strain having selective control effect of moisture-friendly skin microbiome and cosmetic composition comprising the same |
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| US20140065209A1 (en) * | 2011-05-03 | 2014-03-06 | Dupont Nutrition Biosciences Aps | Probiotic bacteria for the topical treatment of skin disorders |
| CN105451751A (en) * | 2012-10-31 | 2016-03-30 | 株式会社爱茉莉太平洋 | Composition for removing keratinous skin material comprising green tea lactobacillus |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140065209A1 (en) * | 2011-05-03 | 2014-03-06 | Dupont Nutrition Biosciences Aps | Probiotic bacteria for the topical treatment of skin disorders |
| CN105451751A (en) * | 2012-10-31 | 2016-03-30 | 株式会社爱茉莉太平洋 | Composition for removing keratinous skin material comprising green tea lactobacillus |
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| Title |
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| HYOUNG-JUNE KIM 等: ""Transcriptome analysis of airborne PM2.5-induced detrimental effects on human keratinocytes"", 《TOXICOLOGY LETTERS》 * |
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| CN117379350A (en) * | 2022-11-04 | 2024-01-12 | 爱恩琵化妆品有限公司 | Method for preparing cosmetic composition containing tea tree extract and cosmetic composition prepared from same |
| CN117379350B (en) * | 2022-11-04 | 2024-03-12 | 爱恩琵化妆品有限公司 | Method for preparing cosmetic composition containing tea tree extract and cosmetic composition prepared from same |
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| CN111315357B (en) | 2023-05-05 |
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| KR20190048613A (en) | 2019-05-09 |
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