KR102359223B1 - Cosmetic composition comprising mangifera indica irwin seed extract - Google Patents

Abstract

The present invention relates to a cosmetic composition or composition for external application for skin containing an apple mango seed extract as an active ingredient, and more particularly, since the apple mango seed extract has antibacterial activity, anti-inflammatory activity and antioxidant activity against skin flora , can be used as a cosmetic composition for anti-aging or skin inflammation improvement or a composition for external application for skin, effectively improving skin inflammation and preventing aging.

Description

Cosmetic composition containing apple mango seed extract as an active ingredient {COSMETIC COMPOSITION COMPRISING MANGIFERA INDICA IRWIN SEED EXTRACT}

The present technology relates to a cosmetic composition containing an apple mango seed extract as an active ingredient.

Recently, as many people are continuously exposed to various environments, interest in health has increased. As consumption of natural antioxidants using natural products increases rather than products with synthetic antioxidants, and as interest in natural cosmetic raw materials increases, the development of cosmetics using natural products is also increasing. Naturalism and various biotech materials are attracting attention as new materials that can effectively respond to the skin.

Oxidation that occurs mainly in our body is the generation of a large amount of active oxygen according to the metabolic process, and most of these active oxygen are decomposed in the body. However, when the balance of the defense system is broken, free radicals are not decomposed but accumulated in the body, causing cellular tissue damage due to oxidative stress, which can cause various diseases. Although active oxygen is a substance necessary for cell growth, when the amount of active oxygen is increased or the function of the removal system is weakened due to environmental pollution, exposure to chemical substances, stress, obesity, etc., it causes oxidative damage and causes various diseases. In addition, sebum secreted from the skin gathers due to external influences to increase the activity of the skin flora, which causes skin inflammation, thereby inducing inflammation and accelerating aging of the skin.

The inflammatory response is a biological defense mechanism for cells in the body to protect the human body from damage to the skin caused by stimuli such as physical shock or bacterial infection. Although various immune cells are involved in the regulation of inflammatory responses in the body, macrophages are known to be involved in homeostasis maintenance by participating in various host responses as well as innate immunity. In particular, enzymes related to the biosynthesis of nitric oxide synthase (NOS) and prostaglandin (PG) are known to play an important role in mediating the inflammatory response. Therefore, NOS, an enzyme that produces nitric oxide (NO) from L-Arginine, and cyclooxygenase (COX), an enzyme involved in synthesizing prostaglandins from arachidonic acid, are inflammatory. It is the main goal in blocking

Mango (Mangifera indica) is classified in the Anacardiaceae, Mangifera, and its scientific name is Mangifera indica. Mango is a dicotyledonous plant, an evergreen tree of the Sumacaceae family, reaching 15 to 18 m in height, and the leaves are like spears and 30 cm in length. As one of the top 30 crops in the world, it is a major crop grown in tropical and subtropical regions worldwide. Mangoes are grown.

There are about 2,000 varieties of mango, and research results have been reported that show many differences in fruit color, flavor, and fiber depending on the variety, and various polyphenol compounds exist in mango. Currently, many studies on the physiological activity of mango peels and leaves have been proven, but studies on physiological activity using mango seeds, which are discarded in large quantities, are insufficient.

Korean Patent No. 10-1872458 (registered on June 22, 2018)

In order to solve the above problems, the present invention provides a cosmetic composition or a composition for external application for skin containing an apple mango (Mangifera indica irwin) seed extract, which is a natural material, as an active ingredient.

The cosmetic composition according to the present invention may contain an apple mango seed extract as an active ingredient.

The composition for external application for skin according to the present invention may contain apple mango seed extract as an active ingredient.

In addition, the composition for inhibiting the growth of skin flora according to the present invention may contain an apple mango seed extract as an active ingredient.

Since the apple mango (Mangifera indica irwin) seed extract according to the present invention has antibacterial activity, anti-inflammatory activity and antioxidant activity against the skin flora causing skin inflammatory diseases, a cosmetic composition or skin for anti-aging or skin inflammation improvement It can be used as a composition for external application, and by using it, it is possible to effectively improve skin inflammation and prevent aging.

In addition, as an extract using natural materials, it can be safely used with few side effects.

1 is a graph measuring electron donating ability for each concentration of apple mango seed extract according to an experimental example of the present invention.
2 is an image showing the result of measuring the antibacterial activity of the apple mango seed extract according to an experimental example of the present invention.
3 is a graph measuring cell viability according to an experimental example of the present invention.
4 is a graph measuring the production rate of nitric oxide (NO) according to an experimental example of the present invention.
5 is a result of confirming the expression of pro-inflammatory factors according to an experimental example of the present invention.

Hereinafter, the present invention will be described in detail.

The present inventors have completed the present invention by discovering various physiologically active characteristics such as antioxidant and anti-inflammatory activity in apple mango (Mangifera indica irwin) seed extract, which is a natural material, and confirming the possibility of apple mango seed extract as a natural cosmetic material.

As used herein, the term "extract" refers to a substance obtained by extracting a component of a natural product regardless of an extraction method, an extraction solvent, an extracted component, or the form of an extract. It may include any material obtainable by processing or processing by the method.

In the present specification, the term "cosmetics" may include functional cosmetics having an effect of preventing or improving skin aging, etc., and the functional cosmetics have specialized functionalities that emphasize physiologically active efficacy and effects, unlike general cosmetics. , refers to cosmetics emphasizing specific efficacy and effects, such as skin whitening and skin wrinkle improvement, as defined in the Cosmetics Act.

The present invention provides a cosmetic composition containing an apple mango (Mangifera indica irwin) seed extract as an active ingredient.

In the cosmetic composition according to the present invention, the apple mango seed is a seed part from which the pulp is removed from the apple mango, and may preferably be a seed of an apple mango (Irwin) native to Jeju Island. The apple mango native to Jeju Island is an improved variety of apple mango produced in the subtropics, and has high adaptability to domestic soil. Compared to foreign products with high fiber content, thick skins and large seeds, the improved Jeju mango has thin skins and small seeds, high sugar content, and excellent flavor. The apple mango seeds may be obtained from apple mango harvested or grown in nature, or purchased commercially.

In the cosmetic composition according to the present invention, the apple mango seed extract may be extracted with water, a C1 to C4 alcohol, or a mixed solvent thereof, preferably a hot water extract or a 60 to 80 wt% ethanol aqueous solution extract, more Preferably, it may be a 70 wt% ethanol aqueous solution extract.

The apple mango seed extract may be extracted according to a method commonly used in the art, for example, hot water extraction, ultrasonic extraction, filtration, reflux extraction, etc., which are performed alone or by using two or more methods It can be performed in combination.

In the cosmetic composition according to the present invention, the apple mango seed extract may have antibacterial activity. More specifically, the apple mango seed extract is Escherichia coli (hereinafter, E. coli ), Staphylococcus aureus ( hereinafter, S. aureus ) and Stephylococcus epidermidis ( Staphylococcus epidermidis ; hereinafter, S. epidermidis ) may have antibacterial activity against one or more skin flora selected from the group consisting of.

As used herein, "antibacterial" or "antibacterial activity" refers to a property that resists microorganisms such as bacteria or fungi, and more particularly, antibiotics and the like, which inhibit the growth or proliferation of the microorganisms.

As used herein, "skin flora" is a microorganism that has maintained a close relationship with a specific tissue of a living body, such as the skin, and protects the skin from harmful bacteria and has a beneficial effect, but when it overgrows or generates harmful substances, resistance In this weak person, it becomes an opportunistic infection and can cause skin inflammatory diseases and the like.

According to an experimental example of the present invention, it was found that the antibacterial activity increased according to the concentration of the apple mango seed ethanol extract, and relatively high antibacterial activity was shown in all strains of E.coli , S.aureus and S.epidermidis . , especially in E. coli showed the highest antibacterial activity.

In the cosmetic composition according to the present invention, the apple mango seed extract may have anti-inflammatory activity. More specifically, the apple mango seed extract may reduce nitric oxide (NO) production or inhibit the expression of cyclooxygenase-2 (COX-2).

As used herein, "anti-inflammatory" refers to suppressing inflammation, wherein the inflammation is the body's immune response to a wound or disease, and oxidative stress such as ultraviolet rays, free radicals, and free radicals are inflammatory factors. It activates and causes various diseases and skin aging.

According to an experimental example of the present invention, cellular NO production was rapidly increased by lipopolysaccharide (hereinafter, LPS), but when treated with an apple mango seed hot water extract or an apple mango seed ethanol extract, concentration-dependently It was found that the NO production rate decreased.

In addition, when the expression level of COX-2 protein was measured, when the expression level of COX-2 protein increased by LPS treatment was treated with the extract, it was found that COX-2 protein expression decreased in a concentration-dependent manner.

In the cosmetic composition according to the present invention, the apple mango seed extract may have antioxidant activity.

As used herein, the term “antioxidant” refers to the action of inhibiting oxidation, and cellular damage and pathological diseases are induced due to oxidative stress in vivo, and is also a direct cause of aging.

According to an experimental example of the present invention, the apple mango seed extract was found to have a high content of polyphenol, an antioxidant, and when the free radical scavenging activity was measured, it was increased in a concentration-dependent manner, thereby having an excellent antioxidant effect. was confirmed.

Since the cosmetic composition according to the present invention has antibacterial activity, anti-inflammatory activity and antioxidant activity as described above, it can be utilized as a cosmetic composition for improving anti-aging or skin inflammation.

As used herein, the term "anti-aging" refers to inhibiting or reversing aging, and as aging progresses, skin constituents change or decrease, and receive oxidative stress due to free radicals and active free radicals, and skin wrinkles Changes in which this increases or decreases in elasticity appear.

Through an experimental example of the present invention, the antibacterial effect on the skin flora causing skin inflammatory diseases was confirmed in the apple mango seed extract, and by confirming that it is also effective in improving skin inflammation and preventing skin aging, the apple mango seed extract is effective The cosmetic composition contained as an ingredient can be used for anti-aging or skin inflammation improvement.

In the cosmetic composition according to the present invention, the apple mango seed extract may be included in an amount of 0.001 to 50 parts by weight based on 100 parts by weight of the total cosmetic, but is not limited thereto.

The cosmetic composition according to the present invention may further include one or more of the ingredients listed in the list of cosmetic raw materials registered with the Food and Drug Administration and the ICID (International Cosmetic Ingredient). For example, the cosmetic composition may include an organic solvent, a solubilizer, a thickener, a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, a fragrance, a surfactant, an emulsifier, a filler, a sequestering agent, a preservative, a vitamin, a blocking agent, It may further comprise one or more adjuvants commonly used in the cosmetic field, such as wetting agents, dyes and pigments, hydrophilic or lipophilic actives, or any other ingredients commonly used in cosmetics.

The vitamin may include a water-soluble or oil-soluble vitamin, and the water-soluble vitamin is a water-soluble vitamin that can be formulated in cosmetics, preferably vitamin B1, vitamin B2, vitamin B6, pyridoxine, pyridoxine hydrochloride, vitamin B12, pantothenic acid, nicotinic acid, and nicotinic acid amide, folic acid, vitamin C, vitamin H, and the like, and more preferably vitamin C, which helps collagen synthesis. In addition, salts thereof (thiamine hydrochloride, sodium ascorbate salt, etc.) or derivatives (ascorbic acid-2-phosphate sodium salt, ascorbic acid-2-phosphate magnesium salt, etc.) may be included, and the water-soluble vitamin is a microorganism transformation method , can be obtained by a conventional method such as a purification method from a culture of a microorganism, an enzymatic method or a chemical synthesis method.

The cosmetic composition according to the present invention may be prepared in any formulation conventionally prepared in the technical field to which the present invention pertains. For example, it may be formulated as a lotion, emulsion, lotion, cream, paste, gel, solution, suspension, oil, wax, pack, powder, foundation, spray, surfactant-containing cleansing, etc., but is not limited thereto. . More specifically, flexible lotion, nourishing lotion, astringent lotion, nourishing cream, massage cream, milk lotion, powder, essence, eye cream, sun lotion, sun cream, makeup primer, makeup base, BB cream, powder foundation, emulsion foundation , cleansing cream, cleansing foam, cleansing water, soap, pack, stick type product, balm type product, spray or powder formulation.

When the cosmetic composition according to the present invention is a cream or gel formulation, animal oil, vegetable oil, wax, paraffin, starch, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be further included as a carrier component. can

When the cosmetic composition is a solution or emulsion formulation, a solvent, solvating agent or emulsifying agent, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, propylene glycol, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid It may further include esters and the like.

When the cosmetic composition is a suspension formulation, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be further included.

When the cosmetic composition is in a powder or spray formulation, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be further included as a carrier component, and in particular, in the case of a spray formulation, additional chlorofluorohydrocarbon , may further include a propellant such as propane/butane or dimethyl ether.

When the cosmetic composition is a surfactant-containing cleansing formulation, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide as carrier components Ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glycerides, fatty acid diethanolamide, vegetable oil, linoline derivatives or ethoxylated glycerol fatty acid esters may be further included.

The cosmetic composition according to the present invention may be used alone or by being applied in duplicate, or it may be used by applying it overlappingly with other cosmetic compositions other than the present invention. In addition, the cosmetic composition according to the present invention can be used according to a conventional method of use, and the number of times of use can be changed according to the skin condition or taste of the user.

The present invention provides a composition for external application for skin for improving anti-aging or skin inflammation, comprising an apple mango (Mangifera indica irwin) seed extract as an active ingredient.

As described above, the apple mango seed extract has antibacterial activity, anti-inflammatory activity and antioxidant activity against the skin flora causing skin inflammatory diseases, so it can be used as a composition for external application for skin for anti-aging or improving skin inflammation.

As used herein, the term "external preparation for skin" refers to a preparation that can be applied directly to the outside of the skin, and may include a cosmetic composition, and the dosage form is an ointment, a warning agent, a patch, a liniment agent, a pasta agent, or a cataplasmase, etc. may include, but is not limited thereto.

In addition, the present invention provides a composition for inhibiting the growth of skin flora containing an apple mango (Mangifera indica irwin) seed extract as an active ingredient.

Hereinafter, examples will be described in detail to help the understanding of the present invention. However, the following examples are merely illustrative of the content of the present invention, and the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those of ordinary skill in the art.

<Example 1> Preparation of apple mango (Mangifera indica irwin) seed extract

Apple mango seeds used in an experimental example according to the present invention were used after removing the apple mango flesh native to Jeju Island, dried at room temperature, and stored at 4°C. About 10 times the weight of the sample was added with water and extracted at 100° C. for 9 hours (Heating Mantle, MS-DM608, MTOP6, Korea), 70% ethanol was added, and stirred at room temperature for 24 hours. The extraction method was repeated three times. The water and ethanol extracts were filtered (vacuum pump, DOA-P704-AC, GAST, USA) using filter paper (Whatman) NO.4 and NO.2 in order, and then, a rotary vacuum evaporator (Rotary Vacuum Evaporator, N-100, EYELA, Tokyo Japan) was concentrated (40° C., speed 4 to 5), and it was freeze-dried and stored in a freezer to be used as a sample for the following experiment.

The following experimental results were expressed as Mean±SD (Standard diviation), and the analyzed experimental data was performed NOVA from the experimental data obtained from the control group and each sample. Tukey range test) was performed to verify significance at the 95% level.

<Experimental Example 1> Confirmation of total polyphenols content

The total polyphenol content of the apple mango seed extract prepared according to an embodiment of the present invention was measured by modifying the Folin-Denis method (Folin-Denis, 1912). 2 ml of apple mango seed extract diluted 10-fold in distilled water and 2 ml of 50% Folin-ciocalteu reagent were mixed, and then left at room temperature for 3 minutes. 2 ml of 10% CaCO₂ was added thereto, mixed, and then left at room temperature for 1 hour, and absorbance was measured at 700 nm.

Table 1 below shows the total polyphenol content of the apple mango seed extract according to the extraction method.

Apple Mango Seeds
Hot Water Extract (TMD) Apple Mango Seeds
Ethanol Extract (TME) total polyphenols
(Total polyphennols)
(mg/g dry)
214.65 ± 0.67
380.09 ± 0.75

Referring to Table 1, as a result of measuring the polyphenol content contained in each extract powder, hot water extract (TMD) of apple mango seed was 214.65±0.67 mg/g, and ethanol of apple mango seed (70% ethanol) The extract (TME) showed a high content of 380.09±0.75 mg/g. It was confirmed that it had a high polyphenol content even when compared with the standard material, tannic acid.

<Experimental Example 2> Confirmation of electron donation ability

The radical scavenging activity for 1,1-diphenyl-2-picryl hydrazyl (1,1-diphenyl-2-picryl hydrazyl; hereafter referred to as DPPH) was measured by modifying the Blois (1958) method. Add 3 ml of 60 μM DPPH (Sigma, USA) to 0.5 ml of each diluted concentration (5, 10, 25, 50, 100, 500, 1,000 μg/ml) sample, vortex for 5 seconds, and then for 15 minutes After standing, absorbance was measured at 517 nm.

1 is a graph measuring electron donating ability for each concentration of apple mango seed extract according to an experimental example of the present invention. Electron donating ability is an experiment used as a measure of the action of active oxygen by active radicals in the human body. Here, Vit-C (vitamin C) denotes a control group, TMD denotes an apple mango seed hot water extract, and TME denotes an apple mango seed ethanol extract, and the values in the graph represent the average of three measurements.

Referring to FIG. 1 , as a result of measuring the electron donating ability of each sample at an initial concentration of 5 μg/ml, the hot water extract (TMD) was 7.45±0.62%, and the ethanol extract (TME) was 16.40±0.88%, and at the concentration of 100μg/ml, It showed an effect of more than 90%. When compared with the control group Vit-C, each extract showed a significant trend, and it was found that all extracts increased in a concentration-dependent manner.

<Experimental Example 3> Antibacterial activity confirmation

The test strain used in the antimicrobial activity experiment according to the present invention is Escherichia coli as a skin flora; hereafter, E.coli , KCTC 1039), Staphylococcus aureus ; S. aureus , KCTC 1621), and Staphylococcus epidermidis (hereinafter, S. epidermidis , KCTC 1917) were purchased from the Korea Research Institute of Biotechnology and Bioresources Center and used. The culture conditions were 37° C., cultured in nutrient broth (nutrient broth, DB, USA), and each strain was inoculated into 7 ml liquid medium and subcultured at regular intervals to activate it.

In order to measure the antibacterial activity of the extract prepared according to Example 1, it was carried out using a paper disc method that can clearly confirm the antibacterial inhibitory activity. Each strain cultured in the plate medium was taken 1 platinum 2 doses, cultured in 7 ml of liquid medium for 18 to 24 hours for activation, and then again inoculated with 0.1 ml of the bacterial solution in 10 ml of liquid medium and cultured for 3 to 6 hours, plate medium 1 The improved number of bacteria was inoculated to about 107 cells and spread evenly. A sterilized filter paper disc (filter paper disc, Tokyo, 8 mm, Japan) is placed on a solid plate medium, and the extract samples are absorbed by concentration so as to be 0.05 ml/disc, and incubated at 37°C for 18 to 24 hours, and then the disc ( The diameter of the clear zone (mm) around the disc) was measured.

Figure 2 is an image showing the measurement result of the antibacterial activity of the apple mango seed extract according to an experimental example of the present invention, Table 2 below is the clear zone diameter measurement result for each strain according to the concentration of the apple mango seed extract treatment, It is expressed as mean±SD of the three measured values.

strain
(strain) density
(mg/ml) Clear zone diameter (mm) Hot Water Extract (WE) Ethanol Extract (EE)

E. coli One - (no clear zone) 9.0±0.0 5 - 10.0±0.0 10 - 11.0±0.9 20 - 13.0±1.1

S. epidermidis One - 8.0±0.0 5 - 9.0±0.7 10 - 9.5±11 20 - 11.5±0.8

S. aureus One - 8.0±0.0 5 - 10.0±0.3 10 - 11.0±0.2 20 - 13.0±1.0

Referring to Figure 2 and Table 2, the hot water extract (WE) did not show activity, but the 70% ethanol extract (EE) showed a tendency to increase the antibacterial activity as the treatment concentration of the extract increased. Relatively high antibacterial activity was mostly confirmed in all strains, and in particular, a high inhibition ring of 9.0 to 13.0 mm was confirmed in E. Coli . Therefore, it was confirmed that the apple mango seed extract according to an embodiment of the present invention has a high potential as an antibacterial material for skin flora causing skin inflammatory diseases.

<Experimental Example 4> Confirmation of cell viability

RAW264.7 cells, a mouse macrophage cell line, were purchased from the Korea Cell Line Bank and obtained from 10% fetal bovine serum (hereinafter, FBS, HyClone, USA), 100U/ml penicillin/streptomycin (penicillin/streptomycin, HyClone, USA). Incubated in an incubator at 37°C and 5% CO₂ conditions in Dulbecco's Modified Eagle Medium (hereinafter, DMEM, HyClone, USA) containing The cells were cultured to grow 80-90%.

When the extract prepared according to Example 1 was exposed to the cells, MTT assay was performed to confirm the viability on the cells. First, in a 96-well plate (well plate), 1×10 ⁵ cells/well was dispensed, and 24 hours later, extract samples dissolved in media were added to suit each concentration. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, MTT] After adding 2.5 mg/ml to each well of 40 μl and reacting for 4 hours in an incubator, the supernatant was removed. Dimethyl sulfoxide (dimethyl sulfoxide, DMSO) was mixed in 100 μl aliquots and completely dissolved, followed by measurement at 540 nm using a spectrophotometer (Benchmark PLUS Bio-Rad).

3 is a graph measuring cell viability according to an experimental example of the present invention. TMD means apple mango seed hot water extract, TME means apple mango seed ethanol extract, and the graph shows the average of three measurements (***p<0.001).

Referring to FIG. 3 , the apple mango seed extract showed a survival rate of 95% or more at a concentration of 100 μg/ml, and a cell viability of 50% or less at a concentration of 500 μg/ml. Accordingly, in subsequent experiments, experiments were carried out at concentrations of 5, 10, 50, and 100 μg/ml.

<Experimental Example 5> Confirmation of nitric oxide (NO) inhibitory activity

Nitric oxide (hereinafter, NO) was measured as nitrite and nitrate in the amount of NO in the supernatant of the cell. A grease reagent, which is a safe form after reduction with nitrate for nitrite, was used, and when the concentration of RAW264.7 cells in a 96-well plate (5×10⁴cells/well) was 80%, a phosphate buffer solution was used. After washing twice with (phosphate buffer saline; hereinafter, PBS), it was cultured for at least 18 hours using a serum-free medium. Thereafter, the extract samples were treated by concentration and cultured for 1 hour, and then treated with 1 μg/ml of lipopolysacchride (hereinafter, LPS) and cultured for 24 hours. After taking the supernatant of the culture solution and reacting it with a grease reagent, the absorbance was measured at 540 nm with an ELISA reader, and the NO production rate was expressed as a percentage.

4 is a graph measuring the production rate of nitric oxide (NO) according to an experimental example of the present invention. TMD means apple mango seed hot water extract, TME means apple mango seed ethanol extract, and the graph shows the average of three measurements (***p<0.001).

Referring to FIG. 4 , when the LPS treatment was performed, the NO production rate of the cells increased rapidly to 6.758 ± 0.18 mM/ml, but when the apple mango seed extract sample was treated, the NO production rate was decreased in a concentration-dependent manner, especially , at 100 μg/ml, the hot water extract showed the greatest decrease to 4.65 ± 0.29 mM/ml and the ethanol extract to 4.32 ± 1.03 mM/ml.

<Experimental Example 6> Confirmation of expression of pro-inflammatory factors

After RAW264.7 cells were suspended in DMEM containing 10% FBS, 3 ml each was dispensed to a cell number of 2×10 ⁶ cells/well in a 6-well plate, and cultured at 37° C., 5% CO ₂ in an incubator for 24 hours. After exchanging with new DMEM medium, the stimulant LPS was treated and cultured for 2 hours, and then the extract was treated and cultured in the cells by concentration. After 24 hours, the medium was removed and washed with cold PBS to measure protein expression, and the cell lysates were extracted by adding Protease & Phosphatase Single-Use inhibitor Cocktail 100X to RIPA buffer. Proteins were quantified (BCA protein kit), electrophoresed on 10% SDS-PAGE, transferred to a membrane, and blocked in 5% skim milk for 1 hour.

Protein expression of COX-2 was confirmed using an enhanced chemiluminescence (ECL) solution (Thermo, USA). As an internal control, beta-actin diluted at 1:1000 (β-actin, Santa Cruz Biotechnology, Santa Cruz, California, USA) was used.

5 is a result of confirming the expression of pro-inflammatory factors according to an experimental example of the present invention.

Referring to FIG. 5, as a result of treating RAW264.7 cells with apple mango seed extract at a concentration of 5-100 μg/ml and measuring the expression level of COX-2 protein, the LPS alone treatment group and each concentration compared to the LPS untreated group The expression level of the protein in the star treatment group increased, and when the extract was treated, it was confirmed that the expression level of COX-2 decreased in a concentration-dependent manner.

As described above in detail a specific part of the content of the present invention, for those of ordinary skill in the art, it is clear that this specific description is only a preferred embodiment, and the scope of the present invention is not limited thereby. do. That is, the substantial scope of the present invention is defined by the appended claims and their equivalents.

Claims (10)

Contains 70% ethanol extract of apple mango (Mangifera indica irwin) seed as an active ingredient,
The concentration of the extract is 75 to 125 μg/ml, and has antibacterial activity, nitric oxide production reduction and COX-2 (cyclooxygenase-2) expression inhibitory activity against Staphylococcus epidermidis, S. epidermidis. A cosmetic composition, characterized in that delete delete delete delete delete The method of claim 1,
The extract is
A cosmetic composition, characterized in that it is included in an amount of 0.001 to 50 parts by weight based on 100 parts by weight of the total cosmetic. The method of claim 1,
The composition is
A cosmetic composition, characterized in that it has the formulation of a softening lotion, nutrient lotion, nutrient cream, essence or pack. delete Contains 70% ethanol extract of apple mango (Mangifera indica irwin) seed as an active ingredient,
The concentration of the extract is 75 to 125 μg/ml, and has antibacterial activity, nitric oxide production reduction and COX-2 (cyclooxygenase-2) expression inhibitory activity against Staphylococcus epidermidis, S. epidermidis. A composition for inhibiting the growth of skin flora, characterized in that.

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