Anti-aging skin care composition with four-resistance effect as well as preparation method, product and application of anti-aging skin care composition

Abstract

The invention belongs to the technical field of cosmetics, and particularly relates to an anti-aging skin care composition with four-resistance effects as well as a preparation method, a product and application of the anti-aging skin care composition. The composition is prepared from ergothioneine, kava pepper and pomegranate fruits. The mass ratio of the ergothioneine to the Kava pepper to the pomegranate fruit is 1: (10-40): (1-4). The composition provided by the invention can be used for protecting skin through multiple ways of resisting oxidation, resisting inflammation, resisting saccharification, resisting light aging and the like, so that the anti-aging effect is realized. The technical scheme provided by the invention not only meets diversified skin care requirements, but also improves the skin care effect, and has great advantages in the anti-aging aspect.

Description

Technical Field

[0001] The present invention belongs to the technical field of cosmetics, and in particular relates to an anti-aging skin care composition with quadruple anti-effects, a preparation method thereof, a product and an application thereof. Background Art

[0002] Skin aging is a complex biological process, which is affected by many factors such as genetics, environment and lifestyle. Aging is divided into endogenous aging and exogenous aging. The factors causing endogenous aging include changes in cell metabolism, changes in hormone levels, oxidative stress response, etc. The most important factor causing exogenous aging is ultraviolet rays, followed by lifestyle, such as high-sugar and high-fat diet, staying up late, improper skin care, etc. Based on these two aspects, the ways to fight skin aging are divided into anti-oxidation, anti-glycation, anti-inflammatory, and anti-photoaging.

[0003] Anti-aging products are numerous and include essences, lotions, creams, masks and other categories of products, but the core of these products is anti-aging ingredients. Commonly added anti-aging ingredients include retinol, retinal substances, peptides, vitamin C, niacinamide, and nicotinamide. Retinol, retinal and niacinamide are highly irritating and sensitive to light, so adding them has certain safety risks; peptides have significant anti-aging effects, but the anti-aging process is slow and they are easily affected by other ingredients, temperature, pH value, etc. and lose their activity; nicotinamide is expensive and slow in action; vitamin C and other antioxidant substances are easily oxidized and their activity is unstable, which affects the anti-aging effect.

[0004] In addition to the anti-aging ingredients that are yet to be discussed, the products in the prior art have a single anti-aging pathway, usually only adding antioxidant ingredients, or only adding anti-ultraviolet ingredients, and the formula design is unreasonable. Skin conditions vary from person to person. Even for the same skin type, the effects of use will be different due to the environment, eating habits, and lifestyle. Some people see results, while others see little effect from anti-aging products. Single-pathway ingredients have limited effects. If anti-aging effects are achieved through single-pathway ingredients, more serious problems will arise. For example, in order to reduce costs, the amount of anti-aging ingredients added is reduced, and the expected anti-aging effect cannot be achieved. For example, some creams claim to contain multiple peptides, but the actual content is extremely low; or in order to ensure the effect, the dosage is increased, thereby ignoring the scientific nature and balance of the overall formula, which may break the natural balance of the skin and cause new skin problems. For example, high-concentration retinol products are prone to skin intolerance.

[0005] Therefore, there is an urgent need to develop a multi-channel skin care composition to meet diverse skin care needs and improve skin care effects and safety. Summary of the invention

[0006] In view of the above-mentioned deficiencies, the present invention provides a multi-pathway skin care composition, which can achieve anti-oxidation, anti-inflammation, anti-glycation and anti-photoaging, and ultimately achieve anti-aging effects through four anti-pathways.

[0007] The technical solution of the present invention is: On the one hand, the present invention provides a composition, comprising ergothioneine, kava and pomegranate fruit; the mass ratio of ergothioneine, kava and pomegranate fruit is 1:10-40:1-4.

[0008] Specifically, the mass ratio of ergothioneine, kava and pomegranate fruit can be 1:10:1, 1:10:2, 1:10:3, 1:10:4, 1:11:1, 1:11:2, 1:11:3, 1:11:4, 1:15:1, 1:15:2, 1:15:3, 1:15:4, 1:20:1, 1:20:2, 1:20:3, 1:20:4, 1:30:1, 1:30:2, 1:30:3, 1:30:4, 1:40:1, 1:40:2, 1:40:3, 1:40:4 and any ratio within the range.

[0009] Preferably, the mass ratio of ergothioneine, kava and pomegranate fruit can be 1:20:2, 1:10:1, 1:40:2 or 1:20:4.

[0010] Specifically, the composition further comprises one or more of gentian, carnosine, rose extract and palmitoyl tripeptide.

[0011] In another aspect, the present invention provides a method for preparing the aforementioned composition, comprising mixing ergothioneine, kava and pomegranate fruit according to a proportion.

[0012] In another aspect, the present invention provides a cosmetic comprising the aforementioned composition.

[0013] Specifically, the cosmetic further comprises one or more of an emulsifier, a moisturizer, a thickener, a chelating agent, a preservative or a pH adjuster.

[0014] Preferably, the emulsifier can be one or more of steareth-2, steareth-21, oleth-10, PEG-30-dipolyhydroxystearate, PEG-100 stearate, oleic acid, linoleic acid, linolenic acid, palmitic acid, behenic acid, stearic acid, lauric acid, polyglycerol esters, methyl glucose sesquistearate, sucrose distearate, sucrose stearate, sucrose cocoate, potassium cetyl phosphate, sodium lauryl sulfate, sodium isostearyl lactylate, sodium stearoyl glutamate, sodium cetearyl sulfate, cetrimonium, stearyl trimethyl ammonium chloride, distearyl dimethyl ammonium chloride, palmitamidopropyl trimethyl ammonium chloride or soy lecithin; The moisturizing agent may be one or more of glycerin, sorbitol, mannitol, xylitol, cetyl alcohol, lanolin alcohol, sodium lactate, sodium pyrrolidone, hexyl laurate, isopropyl myristate, isopropyl palmitate, allantoin or ceramide; The thickener may be one or more of starch, xanthan gum, carrageenan, pectin, sodium alginate, guar gum, gum arabic, carbomer, methylcellulose, hydroxypropyl methylcellulose, hydroxyethyl cellulose or sodium polyacrylate; The chelating agent may be one or more of EDTA, disodium EDTA, nitrilotriacetic acid, tetrasodium glutamate diacetate, caprylhydroxamic acid or sodium phytate; The preservative may be one or more of phenoxyethanol, benzyl alcohol, methylparaben, ethylparaben, propylparaben, butylparaben, sodium benzoate, potassium sorbate or DMDM ​​hydantoin; The pH adjuster may be one or more of citric acid, lactic acid, tartaric acid, glycolic acid, malic acid, salicylic acid, disodium hydrogen phosphate, borax or ammonia water.

[0015] Specifically, the cosmetics also include one or more of gentian, carnosine, rose extract and palmitoyl tripeptide.

[0016] Specifically, the types of cosmetics include but are not limited to: cream, lotion, essence, moisturizing water or facial mask.

[0017] In another aspect, the present invention provides the use of the aforementioned composition or cosmetic, the use comprising: (1) Preparation of antioxidant products; (2) Preparation of anti-inflammatory products; (3) Preparation of anti-glycation products; (4) Preparation of anti-photoaging products; and / or, (5) Preparation of anti-aging products.

[0018] Specifically, the products include but are not limited to: cosmetics or medicines.

[0019] Specifically, the product may further include other active ingredients; the other active ingredients include but are not limited to one or more of gentian, carnosine, rose extract and palmitoyl tripeptide.

[0020] In another aspect, the present invention provides a medicine, which comprises the aforementioned composition Specifically, the medicine also includes pharmaceutically acceptable excipients.

[0021] More specifically, the auxiliary material is selected from one or a combination of two or more of wetting agents, emulsifiers, preservatives, antioxidants, buffers, excipients, diluents, lubricants, antibacterial agents, suspending agents, suspending aids, solubilizers, thickeners, stabilizers, sweeteners and flavors.

[0022] Preferably, the food science or pharmaceutically acceptable excipient can be selected from at least one of lactose, mannose, starch, gum arabic, calcium phosphate, alginate, gelatin, calcium silicate, polyvinyl pyrrolidone, cellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, magnesium stearate and mineral oil.

[0023] The beneficial effects of the present invention are: (1) The composition provided by the present invention has strong antioxidant capacity.

[0024] (2) The composition provided by the present invention has strong anti-inflammatory ability.

[0025] (3) The composition provided by the present invention has excellent anti-sugar effect.

[0026] (4) The composition provided by the present invention has good anti-photoaging ability.

[0027] (5) The composition provided by the present invention has anti-aging effects. DETAILED DESCRIPTION

[0028] The present invention will be further described in detail below by way of examples. The following examples are only a part of the present invention and are not intended to limit the present invention, but are only intended to illustrate the present invention. The experimental methods used in the following examples are all routine experiments unless otherwise specified, and the materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial sources.

[0029] Sources of the products / raw materials of the present invention: See Table 1 for purchase sources: Table 1

[0030] Example 1 The invention provides a composition consisting of ergothioneine, kava and pomegranate fruit.

[0031] The mass ratio of ergothioneine, kava and pomegranate fruit is 1:20:2.

[0032] The preparation method of the composition is: uniformly mix the components according to the above mass ratio to obtain the composition.

[0033] Example 2 The difference from Example 1 is that the mass ratio of ergothioneine, kava pepper and pomegranate fruit is 1:10:1. Others are the same as Example 1.

[0034] Example 3 The difference from Example 1 is that the mass ratio of ergothioneine, kava pepper and pomegranate fruit is 1:40:2. Others are the same as Example 1.

[0035] Example 4 The difference from Example 1 is that the mass ratio of ergothioneine, kava pepper and pomegranate fruit is 1:20:4. Others are the same as Example 1. Comparative Example 1 The difference from Example 1 is that Piper mesenteriae is replaced by Gentiana scabrae in equal weight. The rest is the same as Example 1.

[0036] Comparative Example 2 The difference from Example 1 is that kava pepper is replaced by gentian of equal weight, and pomegranate fruit is replaced by carnosine of equal weight. The rest is the same as Example 1.

[0037] Comparative Example 3 The difference from Example 1 is that pomegranate fruit is replaced with carnosine of equal weight. Other parts are the same as Example 1.

[0038] Comparative Example 4 The difference from Example 1 is that ergothioneine is replaced by ellagic acid in equal parts by weight. Others are the same as in Example 1.

[0039] Comparative Example 5 The difference from Example 1 is that the composition is replaced with a commercially available anti-aging lotion (double anti-aging brightening essence).

[0040] Comparative Example 6 The difference from Example 1 is that kava pepper is replaced with pomegranate fruit of equal weight. The rest is the same as Example 1.

[0041] Comparative Example 7 The difference from Example 1 is that pomegranate fruit is replaced with kava pepper in equal weight. Other parts are the same as Example 1.

[0042] Comparative Example 8 The difference from Example 1 is that kava pepper and pomegranate fruit are not added. Others are the same as Example 1.

[0043] Basic Experimental Example: Determination of Cell Safety Concentration The compositions described in the present invention are all compositions prepared according to Examples 1 to 4 and Comparative Examples 1 to 8.

[0044] The method for determining the safe concentration of cells is as follows: Logarithmic phase cells (macrophages RAW264.7 / human skin fibroblasts HFF1 / human fibroblasts L929) were inoculated in a 96-well plate and cultured for 24 hours, and then divided into a normal control group (cells and culture medium) and a sample group (cells, culture medium and composition). The old solution was discarded, the cells were rinsed once with PBS, the control group was added with culture medium, the sample group was added with sample solution (composition + culture medium), and placed in a 37°C 5% CO2 incubator for 24 hours, the old solution was discarded, the cells were rinsed twice with PBS, 110 μL of culture medium containing 10% v / v CCK-8 solution was added to each well, and a blank group was set up. Only 110 μL of CCK-8 solution was added to the blank group wells. After incubation at 37°C for 2 hours, the absorbance OD at 450 nm was detected by an enzyme reader to calculate the cell survival rate. The highest concentration of the composition with a cell survival rate greater than 95% is the cell safety concentration of the sample.

[0045] The culture medium used for the human skin fibroblast HFF1 and mouse fibroblast L929 is: DMEM high glucose basal medium 85% v / v + fetal bovine serum 15% v / v, both purchased from Gibco, USA; The culture medium used for the macrophage RAW264.7 is: 90% v / v DMEM high-glucose basal medium + 10% v / v fetal bovine serum, both purchased from Gibco, USA.

[0046] Cell survival rate = (OD value of sample group - OD value of blank group) / (OD value of normal control group - OD value of blank group) × 100%.

[0047] Table 2 Maximum safe cell concentration

[0048] The final concentrations of the compositions tested in the cell experiments in the following effect examples except 1.1 and 3.1 were all half of the corresponding concentrations in Table 2.

[0049] Effect Example 1 Evaluation of Antioxidant Capacity 1.1 Free radical scavenging ability determination method (DPPH evaluation method) Take 2.0 mL of the composition, place it in a 10 mL centrifuge tube, add 2.0 mL of DPPH solution, shake vigorously, react at room temperature in the dark for 30 minutes, and use anhydrous ethanol as a blank, add it to a 1 cm cuvette, and measure the absorbance at a wavelength of 517 nm. Calculate the IC50 value based on the standard curve and regression equation.

[0050] 1.2 Method for determining the Nrf2 enhancement factor in skin fibroblasts Logarithmic phase HFF1 cells (Hefei Wanwu Biotechnology Co., Ltd.) were inoculated into a 24-well plate. After overnight culture, the supernatant was discarded, culture medium was added to normal control wells, and culture medium containing the composition was added to sample wells. Culture was continued under different conditions. After 24-48 hours of culture, the cell culture supernatant was discarded, the cells were washed twice with PBS buffer, and after sufficient lysis with lysis buffer, the cell lysate was collected, centrifuged and the supernatant was taken for later use. The Nrf2 content in the cell lysate was determined according to the instructions of the Nrf2 kit (Wuhan Aidikang Biotechnology Co., Ltd., Catalog No. AD32448).

[0051] The culture medium is: DMEM high-glucose basal medium 85% v / v + fetal bovine serum 15% v / v.

[0052] The calculation formula for Nrf2 increase factor is: increase factor = (Nrf2 content in sample wells - Nrf2 content in normal control wells) / Nrf2 content in normal control wells.

[0053] The results are shown in Table 3: Table 3 Evaluation of antioxidant capacity

[0054] Note: The lower the IC50 value of the scavenging rate, the stronger the oxygen free radical scavenging ability and the stronger the exogenous antioxidant capacity; the greater the Nrf2 increase, the stronger the endogenous antioxidant capacity.

[0055] Effect Example 2 Evaluation of Anti-inflammatory Ability 2.1 Method for determining the percentage of NO reduction relative to the model Logarithmic phase RAW264.7 cells (purchased from the National Laboratory Cell Resource Sharing Platform) were inoculated into 24-well plates and cultured for 24 hours. The supernatant was carefully discarded, and normal control wells, LPS-induced model wells (culture medium + 1μg / mL LPS), and sample wells containing the composition (culture medium + 1μg / mL LPS + composition) were set up. The above corresponding solutions were added to each well and cultured for 24 hours. According to the instructions of the nitric oxide kit (purchased from Biyuntian Biotechnology Co., Ltd., item number S0021S), 50μL of culture supernatant was taken from each well, 50μL of reagent 1 was added first, and then 50μL of reagent 2 was added. OD540 was measured, and the NO concentration in each well was calculated according to the standard curve.

[0056] The culture medium is: 90% v / v DMEM high glucose culture medium + 10% v / v fetal bovine serum.

[0057] The calculation formula for the NO reduction percentage is: NO reduction percentage = (NO concentration in model well - NO concentration in sample well) / NO concentration in model well.

[0058] 2.2 Method for determining the percentage reduction of NFκB relative to the model Logarithmic phase RAW264.7 cells were inoculated into 24-well plates and cultured for 24 hours. The supernatant was carefully discarded, and normal control wells (culture medium), LPS-induced model wells (culture medium + 1μg / mL LPS), and sample wells containing the composition (culture medium + 1μg / mL LPS + composition) were set up. The above corresponding solutions were added to each well and cultured for 24 hours. The cell culture supernatant was taken, centrifuged and the supernatant was taken for use. The NF-κB content in the cell lysate was determined according to the instructions of the NF-κB kit (purchased from Wuhan Aidikang Biotechnology Co., Ltd., catalog number AD42446).

[0059] The culture medium is: 90% v / v DMEM high-glucose basal culture medium + 10% v / v fetal bovine serum.

[0060] The calculation formula of NF-κB reduction percentage is: NF-κB reduction percentage = (NF-κB concentration in model well - NF-κB concentration in sample well) / NF-κB concentration in model well.

[0061] The results are shown in Table 4: Table 4 Evaluation of anti-inflammatory ability

[0062] Note: " / " indicates no effect. The model is that RAW264.7 cells are treated with bacterial endotoxin lipopolysaccharide to cause cellular inflammatory response. Compared with the model group, the higher the percentage of NO reduction, the stronger the anti-inflammatory ability; NFκB is activated by lipopolysaccharide, and after sample treatment, the inflammatory response is inhibited. Therefore, the higher the percentage of reduction, the better the anti-inflammatory effect.

[0063] Effect Example 3 Evaluation of Anti-glycation Ability 3.1 Determination of the inhibition rate of AGEs formation in the BSA-fructose model Fructose and BSA were dissolved in phosphate buffer (0.2 mol / L, pH 7.4, containing 0.02%wt proclin 300) to concentrations of 1.5 mol / L and 60 mg / mL, respectively. 500 μL BSA and 500 μL fructose solution were incubated with 500 μL of the composition (1 mg / mL) in a 10 mL centrifuge tube at 50 °C for 24 h as the experimental group. The blank group was replaced with 500 μL deionized water, and the positive control group was replaced with 500 μL aminoguanidine. After incubation, it was diluted 20 times with phosphate buffer, and the fluorescence intensity was detected at an excitation wavelength of 360 nm and an emission wavelength of 460 nm using a fluorescence microplate reader to calculate the amount of AGEs generated. If the sample itself has fluorescence, the background absorption needs to be detected and the final result needs to be corrected. The background absorption only contains the composition, and other solutions are replaced with phosphate buffer.

[0064] The inhibition rate of AGEs production was calculated according to the following formula: inhibition rate (%) = [1-(fluorescence intensity of the experimental group / fluorescence intensity of the blank group)] × 100%.

[0065] 3.2 Determination of anti-glycation ability of skin fibroblasts (1) ROS level reduction rate Take the logarithmic phase L929 cells and inoculate them into a 24-well plate. Set up control wells (medium containing 25mM glucose), model wells (medium containing 270mM glucose), sample wells (medium containing 270mM glucose and combination), and 4 parallels in each group. Treat L929 cells with the corresponding solutions in the above wells for 48h. Dilute the DCFH-DA in the ROS kit (purchased from Biyuntian Biotechnology Co., Ltd., catalog number S0033M) 1000 times with serum-free medium (DMEM high-glucose medium), remove the supernatant in the 24-well plate, and add 500μL of diluted DCFH-DA to each well. Incubate in a cell culture incubator at 37℃ for 20min. Wash the cells three times with serum-free cell culture medium, and detect the fluorescence intensity with a fluorescence microplate reader.

[0066] The culture medium is: DMEM high-glucose basal culture medium 85% v / v + fetal bovine serum 15% v / v, and the control wells and the sample wells and the model wells differ only in the glucose content in the culture medium.

[0067] The formula for calculating the ROS level reduction rate is: ROS level reduction rate = (fluorescence intensity of the model well - fluorescence intensity of the sample well) / (fluorescence intensity of the model well - fluorescence intensity of the control well).

[0068] (2) AGEs generation inhibition rate Logarithmic phase L929 cells were inoculated into 24-well plates, and control wells (medium containing 25mM glucose), model wells (medium containing 270mM glucose), and sample wells (medium containing 270mM glucose and a combination) were set up, with 4 parallels in each group, and L929 cells were treated with the corresponding solutions in the above wells for 48h. The supernatant was discarded, washed once with PBS, 60μL Western and IP lysis buffer was added to each well, lysed on ice for 20min, transferred to a 1.5mL centrifuge tube, centrifuged at 12000rpm and 4℃ for 15min, and the supernatant was taken and detected according to the steps of the AGEs kit (purchased from Shanghai ELISA Biotechnology Co., Ltd., catalog number m1002154). With the concentration of the standard as the horizontal axis and the corresponding OD value as the vertical axis, the linear regression curve of the standard was drawn, and the AGEs concentration value of each sample was calculated according to the curve equation.

[0069] The culture medium is: DMEM high-glucose basal culture medium 85% v / v + fetal bovine serum 15% v / v, and the control wells and the sample wells and the model wells differ only in the glucose content in the culture medium.

[0070] The calculation formula of AGEs generation inhibition rate is: AGEs generation inhibition rate = (AGEs concentration value in model well - AGEs concentration value in sample well) / (AGEs concentration value in model well - AGEs concentration value in control well).

[0071] (3) Increased rate of collagen I Logarithmic phase L929 cells were inoculated into 24-well plates, and control wells (medium containing 25mM glucose), model wells (medium containing 270mM glucose), and sample wells (medium containing 270mM glucose and a combination) were set up, with 4 parallels in each group. L929 cells were treated with the corresponding solutions in the above wells for 48h. The supernatant was taken and tested according to the steps of the collagen I kit (purchased from Shanghai ELISA Biotechnology Co., Ltd., catalog number ml058169). With the concentration of the standard as the horizontal axis and the corresponding OD value as the vertical axis, the linear regression curve of the standard was drawn, and the concentration value of each sample was calculated according to the curve equation.

[0072] The culture medium is: DMEM high-glucose basal culture medium 85% v / v + fetal bovine serum 15% v / v, and the control wells and the sample wells and the model wells differ only in the glucose content in the culture medium.

[0073] The calculation formula of collagen I increase rate is: collagen I increase rate = (collagen I concentration value in sample well - collagen I concentration value in model well) / (collagen I concentration value in control well - collagen I concentration value in model well).

[0074] The results are shown in Table 5: Table 5 Evaluation of anti-glycation ability

[0075] Note: " / " indicates no effect. The higher the inhibition rate of AGEs generation in the BSA-fructose model, the stronger the anti-glycation effect; when high sugar was used to create a cell glycation model, the ROS in the model group increased significantly, the AGEs generation decreased significantly, and the collagen I decreased significantly. Therefore, the higher the percentage of ROS reduction relative to the model group, the higher the inhibition rate of AGEs generation, the higher the percentage of collagen I increase, and the better the anti-glycation effect.

[0076] Effect Example 4: Evaluation of Anti-light Aging Ability Determination method of anti-photoaging ability based on human skin fibroblasts: (1) Activity enhancement rate relative to the UV model group Logarithmic phase HFF1 cells were inoculated into a 96-well plate, and normal control wells (culture medium), model wells (ultraviolet UVA irradiation + culture medium), and sample wells (ultraviolet UVA irradiation + culture medium containing the composition) were set up, with 6 parallels in each group. L929 cells were treated with the corresponding solutions in the above wells for 24 hours, the supernatant was discarded, and PBS buffer was added to each well. The normal group was wrapped with tin foil, and the model group and the sample group were irradiated with UVA for 20 minutes, the supernatant was discarded, and culture medium was added for 24 hours. The supernatant was discarded, and 100 μL of culture medium containing 10% v / v CCK-8 solution was added to each well, incubated for 2 hours, and the OD value of each well at 450nm was detected.

[0077] The culture medium is: DMEM high-glucose basal medium 85% v / v+fetal bovine serum 15% v / v.

[0078] The formula for calculating the activity enhancement rate relative to the UV model group is: Activity enhancement rate = (OD value of sample well - OD value of model well) / (OD value of normal control well - OD value of model well) (2) ROS level reduction rate Logarithmic phase HFF1 cells were inoculated into 24-well plates, and normal control wells (culture medium), model wells (ultraviolet UVA irradiation + culture medium), and sample wells (ultraviolet UVA irradiation + culture medium containing the composition) were set up, with 4 parallels in each group. L929 cells were treated with the corresponding solution in the above wells for 24 hours, the supernatant was discarded, and PBS buffer was added to each well. The normal group was wrapped with tin foil, and the model group and sample group were irradiated with UVA for 20 minutes, the supernatant was discarded, and the culture medium was added for 24 hours. The DCFH-DA in the ROS kit (purchased from Biyuntian Biotechnology Co., Ltd., item number S0033M) was diluted 1000 times with serum-free culture medium, the supernatant in the 24-well plate was removed, and 500 μL of the diluted DCFH-DA was added to each well. Incubate in a cell culture incubator at 37°C for 20 minutes. Wash the cells three times with serum-free cell culture medium, and detect the fluorescence intensity with a fluorescence microplate reader.

[0079] The culture medium is: DMEM high-glucose basal medium 85% v / v+fetal bovine serum 15% v / v.

[0080] The formula for calculating the ROS level reduction rate is: ROS level reduction rate = (fluorescence intensity of the model well - fluorescence intensity of the sample well) / (fluorescence intensity of the model well - fluorescence intensity of the normal control well).

[0081] The results are shown in Table 6: Table 6 Evaluation of light aging resistance

[0082] Note: The UV model group irradiates skin fibroblasts with a certain amount of ultraviolet radiation, causing a significant decrease in cell activity and a significant increase in ROS. Therefore, relative to the model group, the higher the percentage of cell activity enhancement, the higher the percentage of ROS reduction, and the stronger the anti-photoaging effect.

[0083] Effect Example 5: Evaluation of Anti-aging Effect Collagen and elastin determination method: Logarithmic phase HFF1 cells were inoculated into 24-well plates, and normal control wells (culture medium), model wells (UVA irradiation + culture medium), and sample wells (UVA irradiation + culture medium containing the composition) were set up, with 4 parallels in each group. L929 cells were treated with the corresponding solution in the above wells for 24 hours, the supernatant was discarded, and PBS buffer was added to each well. The normal group was wrapped with tin foil, and the model group and sample group were irradiated with UVA for 20 minutes, the supernatant was discarded, and the culture medium was added for 24 hours. The supernatant was taken and detected according to the steps of the collagen and elastin kit (all purchased from Shanghai ELISA Biotechnology Co., Ltd.; type I collagen: item number ml058169; type Ш collagen: item number ml028126; elastin: item number ml106720.). With the concentration of the standard as the horizontal axis and the corresponding OD value as the vertical axis, the linear regression curve of the standard was drawn, and the concentration value of each sample was calculated according to the curve equation.

[0084] The culture medium is: DMEM high-glucose basal medium 85% v / v+fetal bovine serum 15% v / v.

[0085] The formula for calculating the protein enhancement rate is: protein enhancement rate = (sample porin concentration value - model porin concentration value) / (normal control porin concentration value - model porin concentration value).

[0086] The results are shown in Table 7: Table 7 Anti-aging effect evaluation

[0087] Note: After cell aging, the levels of type I and type II collagen and elastin content decrease significantly. Therefore, the higher the percentage of collagen enhancement and the percentage of elastin enhancement, the stronger the anti-aging effect.

[0088] In summary, compared with the comparative example, Examples 1 to 4 have strong antioxidant, anti-inflammatory, anti-glycation and anti-photoaging effects, and the final anti-aging effect is excellent, especially the various effects of Example 1 are the best.

[0089] The above detailed description is a specific description of one of the feasible embodiments of the present invention, and the embodiment is not intended to limit the scope of the present invention. It should be pointed out that any equivalent implementation or change that does not deviate from the present invention should be included in the scope of the technical solution of the present invention. Therefore, the protection scope of the patent of the present invention shall be based on the attached requirements.

Claims

1. A composition, characterized in that The composition comprises ergothioneine, kava and pomegranate fruit; the mass ratio of ergothioneine, kava and pomegranate fruit is 1:10-40:1-4.

2. The composition according to claim 1, characterized in that The mass ratio of ergothioneine, kava and pomegranate fruit is 1:20:2, 1:10:1, 1:40:2 or 1:20:

4.

3. The method for preparing the composition according to claim 1 or 2, characterized in that: The method comprises mixing ergothioneine, kava and pomegranate fruit according to a certain ratio.

4. A cosmetic, characterized in that: The cosmetic comprises the composition according to claim 1 or 2.

5. The cosmetic according to claim 4, characterized in that: The cosmetic may further include one or more of an emulsifier, a moisturizer, a thickener, a chelating agent, a preservative or a pH adjuster.

6. The cosmetic according to claim 5, characterized in that: The emulsifier is one or more of steareth-2, steareth-21, oleth-10, PEG-30-dipolyhydroxystearate, PEG-100 stearate, oleic acid, linoleic acid, linolenic acid, palmitic acid, behenic acid, stearic acid, lauric acid, polyglycerol esters, methyl glucose sesquistearate, sucrose distearate, sucrose stearate, sucrose cocoate, potassium cetyl phosphate, sodium lauryl sulfate, sodium isostearyl lactylate, sodium stearoyl glutamate, sodium cetearyl sulfate, cetrimonium, stearyl trimethyl ammonium chloride, distearyl dimethyl ammonium chloride, palmitamidopropyl trimethyl ammonium chloride or soy lecithin; The moisturizing agent is one or more of glycerin, sorbitol, mannitol, xylitol, cetyl alcohol, lanolin alcohol, sodium lactate, sodium pyrrolidone, hexyl laurate, isopropyl myristate, isopropyl palmitate, allantoin or ceramide; The thickener is one or more of starch, xanthan gum, carrageenan, pectin, sodium alginate, guar gum, gum arabic, carbomer, methyl cellulose, hydroxypropyl methyl cellulose, hydroxyethyl cellulose or sodium polyacrylate; The chelating agent is one or more of EDTA, disodium EDTA, nitrilotriacetic acid, tetrasodium glutamate diacetate, caprylhydroxamic acid or sodium phytate; The preservative is one or more of phenoxyethanol, benzyl alcohol, methylparaben, ethylparaben, propylparaben, butylparaben, sodium benzoate and potassium sorbate or DMDM ​​hydantoin; The pH regulator is one or more of citric acid, lactic acid, tartaric acid, glycolic acid, malic acid, salicylic acid, disodium hydrogen phosphate, borax or ammonia water.

7. The cosmetic according to any one of claims 4 to 6, characterized in that: The types of cosmetics include creams, lotions, essences, moisturizers or masks.

8. Use of the composition according to claim 1 or 2 or the cosmetic according to any one of claims 4 to 7, characterized in that: The applications include: (1) Preparation of antioxidant products; (2) Preparation of anti-inflammatory products; (3) Preparation of anti-glycation products; (4) Preparation of anti-photoaging products; and / or, (5) Preparation of anti-aging products.

9. The use according to claim 8, characterized in that: The product is a cosmetic or a medicine.

10. The use according to claim 8 or 9, characterized in that: The product also includes one or more of gentian, carnosine, rose flower extract and palmitoyl tripeptide.

11. A drug, characterized in that The medicine comprises the composition according to claim 1 or 2.

Images