KR102669645B1 - Cosmetic Composition for stress relief and skin barrier improvement comprising extract of Momordica charantia - Google Patents

Abstract

The present invention relates to a cosmetic composition for relieving stress and improving the skin barrier containing bitter melon extract as an active ingredient. The cosmetic composition of the present invention relieves stress by suppressing the production of cortisol, a stress hormone, and the expression of 11β-HSD1, an enzyme that regulates cortisol production. It is effective in improving the skin barrier damaged by stress as it exerts a restoration effect on the expression of aquaporin-3 (AQP-3) and filaggrin.

Description

Cosmetic composition for stress relief and skin barrier improvement comprising extract of Momordica charantia} containing extract of bitter melon as an active ingredient

The present invention relates to a cosmetic composition for relieving stress and improving the skin barrier containing bitter melon extract as an active ingredient. The cosmetic composition of the present invention shows a stress relieving effect by inhibiting the production of cortisol, a stress hormone, and inhibiting the expression of 11β-HSD1, an enzyme that regulates cortisol production, and aquaporin-3 (AQP-3) and filaggrin. It is effective in improving the skin barrier damaged by stress by promoting the expression of .

Stress refers to an individual's state of tension or reaction that occurs due to a significant amount of physical or psychological strain, and because the stress response is a subjective emotion and varies greatly from person to person, prevention methods are not clearly defined. However, if stress is not properly prevented and responded to, it can not only cause psychological stress such as depression or anxiety, but also cause physiological changes such as hormonal changes, cardiovascular disease, inflammation, and decreased immune function. Stress activates the central nervous system, autonomic nervous system, and hypothalamic-pituitary-adrenal axis (HPA axis), releasing glucocorticoids, adrenocorticotropic hormone (ACTH), and corticotropic releasing factor (CRF). ) increases the secretion of Additionally, activation of the HPA axis suppresses the activity of immune cells and causes a decrease in lymphocytes, thereby lowering immune function. Among them, glucocorticoids play an important role in immunity, growth, development, metabolism, and body homeostasis. They are steroid hormones synthesized from cholesterol and include cortisol, dexamethasone, prednisone, prednisolone, and aldosterone ( aldosterone), etc. Cortisol is the hormone that reacts first in the stress response and is known as a representative stress hormone.

Cortisol is the hormone that reacts first in the stress response and is known as a representative stress hormone. The production of cortisol is regulated by the HPA axis and the activity of an enzyme called 11β-HSD (hydroxysteroid dehydrogenase). Among them, 11β-HSD is an NADPH-dependent enzyme and two types of isoenzymes exist. 11β-HSD 1 is highly expressed in the liver, fat, and muscles and converts inactive cortisone into active cortisol in the body. 11β-HSD 2 is expressed in the kidneys, liver, lungs, etc. and plays a role in oxidizing cortisol to cortisone. The action of these two types of enzymes regulates cortisol homeostasis in the body and is used as an indicator to check the level of stress by checking the level of cortisol. In addition, cortisol is produced in various cells such as keratinocytes, fibroblasts, and melanocytes present in the skin, and an increase in cortisol weakens immunity and inhibits the tight junctions of skin cells, thereby impairing the function of the skin barrier. Damage has been reported. In a recent study, it was confirmed that cortisol content and expression of 11β-HSD 1 increased in stressful situations compared to normal conditions. In addition, transepidermal water loss increased due to a decrease in skin barrier proteins, proving that cortisol affects the skin barrier.

Modern people are subject to various psychological, mental and physical stress situations. As stress causes decreased immunity and various skin functions, interest in ways to relieve stress is increasing. Currently, physical and mental alternative therapies such as aromatherapy and massage using scents are being used to relieve stress, but the development of products that can alleviate skin damage caused by stress is insufficient, so natural products that are safe for the skin are being used to prevent skin damage. Development of materials that can be improved is necessary.

Republic of Korea Patent No. 10-2128974 (registered on June 25, 2020) describes a cosmetic composition for protecting skin against ultraviolet rays containing complex extracts of bitter melon, persimmon leaf and silkworm as active ingredients. Republic of Korea Patent No. 10-2207995 (registered on January 20, 2021) describes an antibacterial, anti-inflammatory, and skin moisturizing composition containing fermented bitter melon extract as an active ingredient.

The present invention seeks to provide a cosmetic composition that contains bitter melon extract as an active ingredient and has the effect of relieving stress and improving the skin barrier damaged by stress.

The present invention provides a cosmetic composition for improving skin barrier damage containing ethanol extract of Momordica charantia as an active ingredient.

The present invention provides a cosmetic composition for improving skin stress containing ethanol extract of Momordica charantia as an active ingredient.

In the present invention, the extract may be an extract obtained from one or more parts selected from the flowers, leaves, fruits, stems, or roots of bitter melon.

In the present invention, the extract may be characterized in that it is extracted through any one extraction method selected from solvent extraction, supercritical extraction, and ultrasonic extraction.

The present invention provides a cosmetic composition that is effective in relieving stress and improving the skin barrier collapsed due to stress by containing a bitter melon extract that exerts the effect of suppressing cortisol production or expression of 11β-HSD1 and promoting the expression of AQP-3 or filaggrin.

Figure 1 shows the results of confirming the cytotoxicity of bitter melon extract on human-derived keratinocytes.
Figure 2 shows the results of confirming the inhibitory effect of bitter melon extract on cortisol production.
Figure 3 shows the results confirming the inhibitory effect of bitter melon extract on 11β-HSD1 expression.
Figure 4 shows the results confirming the effect of the bitter melon extract on restoring aquaporin-3 (AQP-3) expression.
Figure 5 shows the results confirming the effect of bitter melon extract on restoring filaggrin expression.

The present invention provides a cosmetic composition for improving skin barrier damage containing ethanol extract of Momordica charantia as an active ingredient.

The present invention provides a cosmetic composition for improving skin stress containing ethanol extract of Momordica charantia as an active ingredient.

Bitter melon ( Momordica charantia ) is an annual vine crop that grows in tropical regions such as Asia and Africa and has been mainly used for ornamental purposes. Bitter melon has a bitter taste, so it is also called bitter gourd and is used for edible and medicinal purposes. Bitter melon is known to have high antioxidant and blood sugar control effects as it contains various bioactive ingredients such as vitamin C, vitamin A, amino acids, galactronic acid, pectin, saponin, and β-carotene. In particular, charantin contained in fruits and seeds has been reported to have the effect of strengthening blood sugar levels by promoting insulin secretion. In addition, it is known to have antibacterial and anti-inflammatory effects against viruses and fungi.

Meanwhile, stress can cause not only psychological effects such as depression, sleep disorders, and anxiety, but also decreased immunity, skin inflammation, oxidative stress, and damage to barrier function. Cortisol is the first hormone produced in stressful situations, and its production is regulated by 11β-HSD enzyme activity. It is known that as cortisol increases due to stress, it weakens the skin barrier by reducing factors involved in cell junctions.

Meanwhile, in the present invention, the skin barrier damage may be due to increased cortisol production due to stress.

Meanwhile, the ethanol extract of bitter melon of the present invention may exert an effect of suppressing cortisol production or suppressing the expression of 11β-HSD1 (hydroxysteroid dehydrogenase 1), an enzyme that regulates cortisol production. In addition, the ethanol extract of bitter melon of the present invention may exert the effect of strengthening the skin barrier, which is reduced due to stress, and restoring the expression of aquaporin-3 (AQP-3) and filaggrin, which are factors related to skin moisturization. As a result of an experiment using the ethanol extract of bitter melon of the present invention in an experimental example of the present invention, the effect of suppressing the production of cortisol, a stress hormone, and the effect of suppressing the expression of 11β-HSD1, an enzyme that regulates cortisol production, were confirmed. In addition, the effect of restoring the expression of aquaporin-3 (AQP-3) and filaggrin, factors related to skin barrier strengthening and skin moisturization, which were reduced due to stress, was confirmed. Through this, it was found that the ethanol extract of bitter melon of the present invention has an excellent effect in relieving stress and improving the skin barrier broken down by stress.

Therefore, the cosmetic composition of the present invention has any one effect selected from inhibiting the production of cortisol, inhibiting the expression of 11β-HSD1, increasing the expression of aquaporin-3 (AQP-3), or increasing the expression of filaggrin. It can be something to have.

In the present invention, the ethanol extract of Yeoju extract can be extracted using any one extraction method selected from solvent extraction, supercritical extraction, or ultrasonic extraction. However, extraction is preferably performed using 60-80% (v/v) ethanol as the extraction solvent, and more preferably, extraction is performed using 70% (v/v) ethanol as the extraction solvent.

Meanwhile, in the present invention, the ethanol extract of bitter melon may be characterized as being an extract obtained from any one or more parts selected from the flowers, leaves, fruits, stems, or roots of bitter melon.

In addition, the ethanol extract of bitter melon is preferably contained in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition. More preferably, it is contained in 0.01 to 10% by weight based on the total weight of the cosmetic composition. If the content of bitter melon extract is less than 0.0001% by weight, the effect of relieving skin stress or improving the skin barrier is minimal, and if it exceeds 30.0% by weight, a clear increase in effect may not be seen as the content increases.

Meanwhile, the ingredients included in the cosmetic composition of the present invention may include ingredients commonly used in cosmetic compositions in addition to the ethanol extract of Yeoju of the present invention as an active ingredient, such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and Contains conventional auxiliaries such as flavorings, and carriers.

The cosmetic composition of the present invention can be prepared in any formulation commonly prepared in the art, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing products. , may be formulated into oils, packs, massage creams, sprays, etc., but are not limited thereto. More specifically, it can be manufactured in the form of softening lotion, nourishing lotion, nourishing cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray, or powder.

When the formulation of the cosmetic composition of the present invention is a paste, cream or gel, the carrier ingredient may include animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide. This can be used.

When the formulation of the cosmetic composition of the present invention is a solution or emulsion, a solvent, solubilizing agent, or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, and propylene. These include fatty acid esters of glycol, 1,3-butylglycol oil, glycerol aliphatic esters, polyethylene glycol or sorbitan.

When the formulation of the cosmetic composition of the present invention is a suspension, the carrier component includes water, a liquid diluent such as ethanol or propylene glycol, and a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester. , microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant, etc. can be used.

When the formulation of the cosmetic composition of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder can be used as the carrier ingredient. In particular, when the cosmetic composition is a spray, chlorofluorohydride may be additionally used. May contain propellants such as carbon, propane/butane or dimethyl ether.

When the formulation of the cosmetic composition of the present invention is a surfactant-containing cleansing agent, the carrier ingredients include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, Fatty acid amide ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester can be used.

When the cosmetic composition of the present invention is a soap, a surfactant-containing cleansing formulation, or a surfactant-free cleansing formulation, it can be applied to the skin and then wiped off, removed, or washed with water. As a specific example, the soap is liquid soap, powdered soap, solid soap, and oil soap, the surfactant-containing cleansing formulation is cleansing foam, cleansing water, cleansing towel, and cleansing pack, and the surfactant-free cleansing formulation is cleansing cream. , cleansing lotion, cleansing water, and cleansing gel, but are not limited thereto.

Hereinafter, the configuration of the present invention will be described in detail through the following examples and experimental examples. However, the scope of the present invention is not limited to the following examples and experimental examples, and includes modifications of the technical idea equivalent thereto.

[Example 1: Preparation of bitter melon extract]

After the bitter melon ( Momordica charantia ) was dried in the shade and crushed, 20 times the weight of 70% (v/v) ethanol was added and extracted at room temperature for 24 hours. Afterwards, the extract was filtered, concentrated under reduced pressure, and freeze-dried to prepare a bitter melon extract.

[Experimental Example 1: Confirmation of cytotoxicity of bitter melon extract]

In this experiment, in order to confirm the cytotoxicity of the bitter melon extract, the effect of the bitter melon extract on human-derived keratinocytes was confirmed.

Human-derived keratinocytes were distributed at 2 × 10 ⁴ cells/well in a 96 well plate, and the cells were stabilized for one day in an incubator at 37°C and 5% CO ₂ . Afterwards, the bitter melon extract prepared in Example 1 was diluted to concentrations of 10, 30, and 50 μg/ml and reacted for 24 hours. 20 μl of MTT reagent at a concentration of 5 mg/ml was added to each well and reacted in an incubator for 2 hours. All supernatants were removed, 100 ㎕ of DMSO was added to each well to completely lyse the cells, and the absorbance was measured at 550 nm. Meanwhile, cell survival rate was calculated as a percentage compared to untreated cells and is shown in Table 1 below.

sample Cell viability (%) Untreated 100 Example 1 (㎍/㎖)
(Bitter melon extract) 3 102.35 10 103.51 30 103.32 50 101.77

As a result of the experiment, as shown in Table 1 and Figure 1, the bitter melon extract of the present invention did not show cytotoxicity up to a concentration of 50 μg/ml. Meanwhile, Figure 1 is a graph showing the results of confirming the cytotoxicity of the bitter melon extract.

[Experimental Example 2: Confirmation of the inhibitory effect of bitter melon extract on cortisol production]

In this experiment, we sought to determine the effect of bitter melon extract on cortisol production.

450 ㎕ of human-derived keratinocytes were dispensed into a 24-well plate at 5 × 10 ⁴ cells/well, and the cells were stabilized in an incubator at 37°C and 5% CO ₂ for one day. After providing a stress stimulus by treating with 100 μM of cortisone, the bitter melon extract prepared in Example 1 was diluted to concentrations of 3, 10, 30, and 50 μg/ml, respectively, and reacted for 24 hours. The medium was removed, washed three times with 1×PBS, and the cells were collected using a scraper and sonicated to lyse the cells. The supernatant was recovered by centrifugation at 12,000 rpm and 4°C for 10 minutes, and the intracellular cortisol content was measured using a human cortisol elisa kit. Table 2 below shows the inhibition rate of cortisol production calculated as a percentage.

sample Cortisol production inhibition rate (%) Cortisone (μM) 100 0 Example 1 (㎍/㎖)
(Bitter melon extract) 3 8.53 10 28.18 30 44.94 50 69.14

As a result of the experiment, as shown in Table 2 and Figure 2, the bitter melon extract of the present invention suppressed the production of cortisol in a concentration-dependent manner. In addition, a significant inhibition rate of 28.18% was observed at a concentration of 10 μg/ml, and a high cortisol production inhibition effect of up to 69.14% was confirmed at a concentration of 50 μg/ml (Figure 2). Meanwhile, Figure 2 is a graph showing the results of confirming the inhibitory effect of bitter melon extract on cortisol production.

[Experimental Example 3: Inhibitory effect of 11β-HSD1 expression of bitter melon extract]

In this experiment, to confirm the stress-relieving effect of bitter melon extract, its effect on 11β-HSD1, an enzyme that regulates cortisol production in keratinocytes, was confirmed.

Human-derived keratinocytes were distributed in a 12-well plate at 3 × 10 ⁵ cells/well, and the cells were stabilized for one day in an incubator at 37°C and 5% CO ₂ . The Yeoju extract prepared in Example 1 was diluted and treated at concentrations of 10, 30, and 50 μg/ml. After reacting in an incubator at 37°C and 5% CO ₂ for 6 hours, proteins were separated. The supernatant was removed, washed once with 1 × PBS, and cells were collected by scraping with a scraper. Proteins were separated using cell lysis buffer, proteins were quantified using BCA assay, and a certain amount of proteins were electrophoresed on a 10% SDS-PAGE gel. The separated proteins on the gel were transferred to a PVDF membrane and blocked for 1 hour using 5% skim milk. After washing three times with 1 Afterwards, it was washed three times with 1 × TTBS, reacted with ECL solution, and the band was detected with a Chemi Doc instrument. It was quantified using Image J 1.47 software and quantified based on the expression of beta-actin. Meanwhile, Table 3 below shows the calculated inhibition rate (%) of 11β-HSD1 expression compared to untreated cells.

sample 11β-HSD1 expression inhibition rate (%) Example 1 (㎍/㎖)
(Bitter melon extract) 10 52.47 30 56.62 50 58.49

As a result of the experiment, as shown in Table 3 and Figure 3, the bitter melon extract of the present invention suppressed the expression of 11β-HSD1 in a concentration-dependent manner. In addition, it was confirmed that a significant inhibition rate of 52.47% was observed at a concentration of 10 μg/ml, and that a high 11β-HSD1 expression inhibition effect of up to 58.49% was observed at a concentration of 50 μg/ml. Meanwhile, Figure 3 shows the results confirming the inhibitory effect of bitter melon extract on 11β-HSD1 expression.

[Experimental Example 4: AQP-3 expression recovery effect of bitter melon extract]

In this experiment, in order to confirm the effect of the bitter melon extract on improving the skin barrier damaged by stress, the effect on aquaporin-3 (AQP-3), a factor involved in strengthening the skin barrier, was confirmed.

Keratinocytes were distributed in a 6-well plate at 4 × 10 ⁵ cells/well, and the cells were stabilized in an incubator at 37°C and 5% CO ₂ for one day. After providing stress stimulation by treating with 100 μM of cortisone, the bitter melon extract prepared in Example 1 was diluted to concentrations of 10, 30, and 50 μg/ml, respectively, and reacted for 6 hours. The medium was removed, washed once with 1 × PBS, and cells were collected by scraping with a scraper. RNA was isolated using the Trizol method. Cells were lysed by adding 800 ㎕ of Trizol and reacted at room temperature for 5 minutes, then 200 ㎕ of chloroform was added, vortexed, and reacted at room temperature for 5 minutes. Centrifuge at 12,000 rpm, 4°C for 15 minutes, transfer only the supernatant to a new tube, add the same amount of isopropanol, mix 1 to 3 times, and react at room temperature for 10 minutes. After centrifugation at 12,000 rpm and 4°C for 15 minutes and removing all supernatants, 800 ㎕ of cold 70% ethanol was added to wash them, and centrifugation was performed again at 12,000 rpm and 4°C for 15 minutes. The supernatant was completely removed, the pellet was dried until it became transparent, and DEPC DW was added according to the amount of RNA. RNA was quantified using Nanodrop, PCR was performed, and the band was detected by electrophoresis on a 2% agarose gel. The bands were quantified using Image J 1.47 software and quantified based on the expression of beta-actin. Meanwhile, Table 4 below shows the calculated expression recovery rate (%) of aquaporin-3 (AQP-3) compared to the cortisone treatment group.

sample AQP-3 expression recovery rate (%) Example 1 (㎍/㎖)
(Bitter melon extract) 10 0 30 86.92 50 107.84

As a result of the experiment, as shown in Table 4 and Figure 4, the expression of AQP-3 was significantly suppressed by cortisone treatment, and the bitter melon extract of the present invention restored the expression of AQP-3 reduced by cortisone treatment in a concentration-dependent manner. I ordered it. In addition, it was confirmed that a significant AQP-3 recovery effect of 86.92% was observed at a concentration of 30 μg/ml, and that the expression of AQP-3 was promoted by up to 107.84% at a concentration of 50 μg/ml. Meanwhile, Figure 4 shows the results confirming the effect of recovering the expression of aquaporin-3 (AQP-3) reduced by cortisone treatment of bitter melon extract.

[Experimental Example 5: Restoration effect of filaggrin expression of bitter melon extract]

In this experiment, in order to confirm the effect of the bitter melon extract on improving the skin barrier damaged by stress, the effect on filaggrin, a factor involved in strengthening the skin barrier, was confirmed.

Keratinocytes were distributed in a 6-well plate at 4 × 10 ⁵ cells/well, and the cells were stabilized in an incubator at 37°C and 5% CO ₂ for one day. After providing stress stimulation by treating with 100 μM of cortisone, the bitter melon extract prepared in Example 1 was diluted to concentrations of 10, 30, and 50 μg/ml, respectively, and reacted for 6 hours. The medium was removed, washed once with 1 × PBS, and cells were collected by scraping with a scraper. RNA was isolated using the Trizol method. Cells were lysed by adding 800 ㎕ of Trizol and reacted at room temperature for 5 minutes, then 200 ㎕ of chloroform was added, vortexed, and reacted at room temperature for 5 minutes. Centrifuge at 12,000 rpm and 4°C for 15 minutes, transfer only the supernatant to a new tube, add the same amount of isopropanol, mix 1 to 3 times, and react at room temperature for 10 minutes. It was centrifuged at 12,000 rpm and 4°C for 15 minutes, and all supernatants were removed, washed by adding 800 ㎕ of cold 70% ethanol, and centrifuged again at 12,000 rpm and 4°C for 15 minutes. The supernatant was completely removed, the pellet was dried until it became transparent, and DEPC DW was added according to the amount of RNA. RNA was quantified using Nanodrop, PCR was performed, and the band was detected by electrophoresis on a 2% agarose gel. The bands were quantified using Image J 1.47 software and quantified based on the expression of beta-actin. Meanwhile, Table 5 below shows the calculated recovery rate (%) of filaggrin expression compared to the cortisone treatment group.

sample Filaggrin expression recovery rate (%) Example 1 (㎍/㎖)
(Bitter melon extract) 10 93.03 30 220.66 50 220.99

As a result of the experiment, as shown in Table 5 and Figure 5, the expression of filaggrin was significantly suppressed by cortisone treatment, and the bitter melon extract of the present invention restored the expression of filaggrin reduced by cortisone treatment in a concentration-dependent manner. In addition, it was confirmed that the effect of promoting filaggrin was as high as 93.03% at a concentration of 10 μg/ml, and that the expression of filaggrin was promoted by up to 220.99% at a concentration of 50 μg/ml. Meanwhile, Figure 5 shows the results confirming the effect of restoring the expression of filaggrin, which was reduced by cortisone treatment of bitter melon extract.

In this way, the bitter melon extract of the present invention inhibits the production of cortisol, a stress hormone, suppresses the expression of 11β-HSD1, an enzyme that regulates cortisol production, and inhibits aquaporin-3 (AQP-3) and filaggrin, which are skin barrier factors reduced due to stress. It was confirmed that the recovery effect of filaggrin relieves stress and improves the skin barrier damaged by stress.

Claims (4)

Contains bitter melon ( Momordica charantia ) ethanol extract as an active ingredient,
The extract was extracted at room temperature using 60-80% (v/v) ethanol as an extraction solvent, and inhibited the expression of 11β-HSD1, an enzyme that regulates cortisol production and cortisol production, and aquaporin-3 (aquaporin-3) reduced due to stress. A cosmetic composition for improving skin stress and improving skin barrier damage caused by stress, characterized in that it promotes the expression of AQP-3) and filaggrin.
delete According to paragraph 1,
The extract is,
A cosmetic composition characterized in that it is an extract obtained from any one or more parts selected from the flowers, leaves, fruits, stems, or roots of bitter melon. delete