A composition having an anti-aging effect and use thereof

Abstract

The application belongs to the technical field of cosmetics, and discloses a composition with an anti-aging effect and application thereof; the composition comprises curcumin, galangal extract and piper methysticum extract; the application mainly uses the optimal proportioning combination of curcumin, galangal extract and piper methysticum extract to achieve the synergistic effect of multiple functions. Test results show that the composition is beneficial to delaying aging, can effectively inhibit the glycosylation reaction of the skin to play an anti-aging activity, and has a good soothing and repairing effect on the glycosylation damage caused by ultraviolet rays. The composition can also inhibit the activity of cell tyrosinase to achieve a whitening effect; in addition, the composition can improve the adhesion function of skin epidermal cells, restore and promote the synthesis of collagen, has the effects of tightening the skin and enhancing the skin barrier, has the advantages of being easy to prepare and mixed with other components to be added into cosmetics.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of cosmetics, and particularly relates to a composition with anti-aging effect and application thereof. BACKGROUND

[0002] Skin aging is a complex, multifactorial, and integrated biological process, and the main clinical manifestations are skin sagging, increased wrinkles, dryness and desquamation, yellowing and darkening, and increased pigmented spots. According to different causes of aging, it can be divided into endogenous aging and exogenous aging. Endogenous aging, also known as natural aging, is caused by the slowing down of skin metabolism, the accumulation of harmful substances, and the weakening of skin repair and renewal ability. Exogenous aging is caused by external factors such as ultraviolet radiation, smoking, and exposure to harmful substances. Glycation and ultraviolet radiation are the main causes of exogenous aging in daily life. With the development of society, people consume more types of high-sugar and high-fat food and drinks, which causes glycation in the skin. Glycation is the reaction of free amino groups of macromolecules such as proteins and DNA with aldehyde groups of reducing sugars to produce end products that are not easily degraded, which is called "advanced glycation end products (AGEs)". The accumulation of AGEs in the skin not only damages collagen and elastin in the skin, making the skin lose elasticity, produce wrinkles, and have dark and rough skin. In addition, modern life makes people exposed to the outdoors for a long time, and various digital environments such as mobile phones and computers. Long-term exposure to long-wave ultraviolet radiation (UVA), medium-wave ultraviolet radiation (UVB), and blue light can also damage the structure of the skin, leading to increased glycation in the skin, rapid increase in AGEs content, and skin aging. Therefore, anti-aging cosmetic products are increasingly popular in the market.

[0003] With the continuous penetration of the concept of "green" and "healthy aging" into the cosmetics industry, various types of anti-aging products have appeared on the market. The current mainstream concept is to add antioxidants to eliminate reactive oxygen species (ROS) and combat the generation of free radicals, such as vitamin C, vitamin E, and glutathione. Anti-glycation and anti-photoaging are also being popularized to the public, and some effective ingredients such as collagen and boswellia are gradually added to skincare products. Due to the advantages of plant extracts, such as being derived from nature and having few side effects, natural plant extracts have been widely used as active ingredients in the production of anti-aging cosmetics. Research has found that many antioxidants also have anti-glycation effects, such as glutathione and resveratrol, but the anti-glycation effect is generally not good. There are not many plant extracts that have been developed and applied to anti-glycation and anti-aging cosmetics, and the plant active ingredients added have single anti-glycation effects and do not have synergistic effects, making it difficult to achieve the desired anti-aging effect. Therefore, it is necessary to find anti-aging compositions with clear effects and good anti-glycation effects. This discovery and research may help to find new anti-aging and skin care methods. SUMMARY

[0004] The present application provides a curcumin and zedoary plant composition with anti-aging effect.

[0005] The present application provides the use of the anti-aging composition of the first aspect.

[0006] The present application also provides a product.

[0007] In order to achieve the above-mentioned purpose, the technical scheme adopted by the present application is:

[0008] An anti-aging composition, characterized in that it comprises curcumin, galangal extract and cardamom extract.

[0009] The composition, characterized in that it comprises the following components in mass fraction: 0.1-8 parts of curcumin, 0.1-8 parts of galangal extract and 0.1-5 parts of cardamom extract.

[0010] The composition, characterized in that it comprises the following components in mass fraction: 0.1-4 parts of curcumin, 1-5 parts of galangal extract and 1-4 parts of cardamom extract.

[0011] The composition, characterized in that the galangal extract and the galangal extract are obtained by the following steps: taking 200g of dried rhizomes of galangal and 200g of dried fruits of cardamom as raw medicinal materials, crushing them into powders with a pulverizer, adding ethanol to obtain extracts of the two medicinal materials respectively by ultrasonic, obtaining different concentrations of ethanol eluents of the two medicinal material extracts by using D101 macroporous resin, taking the 60%-90% ethanol eluted part to dry, obtaining extract, and storing.

[0012] The use of the anti-aging composition in the preparation of a cosmetic product or a food additive.

[0013] The use, characterized in that the cosmetic product comprises facial cleanser, cosmetic water, emulsion, facial mask, cream and essence. A cosmetic product, characterized in that it comprises the anti-aging composition of any one of claims 1 to 3 and cosmetically acceptable adjuvant.

[0014] The cosmetic product, characterized in that the adjuvant comprises at least one of emollient, emulsifier, thickening agent, humectant, pH regulator, skin protectant and preservative.

[0015] Curcumin is a polyphenolic compound isolated from turmeric, which has anti-aging effect, can inhibit inflammatory response, and has whitening effect as a plant polyphenol; curcumin can also capture adducts formed by the addition reaction of dicarbonyl compounds (such as MGO) to the terminal reducing amino group of macromolecules during glycosylation to inhibit the generation of AGEs intermediates and finally achieve the effect of anti-glycation; in addition, curcumin is an important intracellular antioxidant that can scavenge reactive oxygen species (ROS) and a series of oxidative stress reactions, and can be applied to various cosmetics; although curcumin is considered a relatively safe natural compound, it is rich in yellow color, has deep color, is difficult to clean at increased dosage, and has poor solubility, which limits its application in the field of anti-aging cosmetics.

[0016] Rhizoma Alpiniae Officinarum Extract is a natural plant extract, which is pungent and hot in nature, and is used for warming middle energizer, dispelling cold, regulating qi and relieving pain.

[0017] Exocarpium Amomi Rotundus Extract is a natural plant extract, which is warm in nature, can dispel stagnated qi, eliminate upper phlegm, and treat wind-cold pathogen in stomach. Preferably, the anti-aging composition comprises the following components in parts by weight: 0.1-8 parts of curcumin, 0.1-8 parts of Rhizoma Alpiniae Officinarum Extract, and 0.1-5 parts of Exocarpium Amomi Rotundus Extract.

[0018] Preferably, the anti-aging composition comprises the following components in parts by weight: 0.5-4 parts of curcumin, 2-5 parts of Rhizoma Alpiniae Officinarum Extract, and 1-4 parts of Exocarpium Amomi Rotundus Extract.

[0019] Preferably, the anti-aging composition comprises the following components in parts by weight: 1-3 parts of curcumin, 2-4 parts of Rhizoma Alpiniae Officinarum Extract, and 2-3.6 parts of Exocarpium Amomi Rotundus Extract.

[0020] Preferably, the anti-aging composition comprises the following components in parts by weight: 2 parts of curcumin, 3 parts of Rhizoma Alpiniae Officinarum Extract, and 3 parts of Exocarpium Amomi Rotundus Extract.

[0021] Preferably, the Rhizoma Alpiniae Officinarum Extract is a Rhizoma Alpiniae Officinarum alcohol extract.

[0022] Preferably, the Exocarpium Amomi Rotundus Extract is an Exocarpium Amomi Rotundus fruit alcohol extract.

[0023] In a second aspect of the present application, a preparation method of the anti-aging composition of the first aspect is provided, which comprises mixing curcumin, Rhizoma Alpiniae Officinarum Extract and Exocarpium Amomi Rotundus Extract.

[0024] In a third aspect of the present application, the anti-aging composition of the first aspect is used for preparing a product.

[0025] Preferably, the anti-aging composition according to the above embodiments of the second aspect of the present application is used in the preparation of cosmetics. Preferably, the cosmetics include creams, eye creams, serums, masks, lotions and sunscreens.

[0026] Preferably, the product has any one of the functions (1) to (5):

[0027] (1) Anti-glycation;

[0028] (2) Soothing repair;

[0029] (3) Whitening;

[0030] (4) Inhibition of tyrosinase activity;

[0031] (5) Repair of skin barrier;

[0032] (6) Anti-aging.

[0033] In summary, the present application has the following advantages and beneficial effects:

[0034] 1. The present application uses a golden ratio of curcumin, galangal extract and cardamom extract as an anti-aging composition, which has a synergistic effect from multiple levels: 1) inhibits skin glycation, regulates the expression of a series of related genes in the glycation process, and has a soothing and repairing effect on the skin; 2) inhibits cell tyrosinase activity, reduces melanin production in the skin aging process from the root, and achieves whitening effect; 3) enhances skin cell adhesion, reduces collagen loss, and repairs skin barrier; 4) repairs skin glycation damage caused by ultraviolet radiation, and has a comprehensive anti-aging effect.

[0035] 2. The anti-aging composition of the present application combines curcumin, galangal extract and cardamom extract to carry out a number of experiments, and it is found through verification that the three alone and in pairs have good anti-glycation and anti-aging effects, and the combination of the three has a more significant effect than the former, and enhances the effect of each other to achieve a synergistic anti-aging effect.

[0036] 3. The anti-aging composition of the present application uses the three in combination, which on the one hand improves the problem of insufficient anti-aging effect caused by the deep color of curcumin and low solubility; on the other hand, the type of components is simple, easy to prepare, and easy to mix with other components into cosmetics, which has the obvious advantage of being easy to add to various types of formulations, so it has great value and prospect in the development of cosmetics.

[0037] Other features and advantages of the present application will be set forth in the specification, and will become apparent from the specification, or will be learned by practice of the present application. BRIEF DESCRIPTION OF DRAWINGS

[0038] Figure 1 Figure showing the effect of the examples and comparative examples on the survival rate of HaCat cells treated with MGO.

[0039] Figure 2 Figure showing the effect of the examples and comparative examples on the expression of the inflammatory factor IL-6 mRNA in HaCat cells treated with MGO.

[0040] Figure 3 Figure showing the effect of the examples and comparative examples on the expression of the inflammatory factor IL-8 mRNA in HaCat cells treated with MGO.

[0041] Figure 4 Figure showing the effect of the examples and comparative examples on the expression of AGER mRNA in HaCat cells treated with MGO.

[0042] Figure 5 Figure showing the effect of the examples and comparative examples on the expression of MMP-9 mRNA in HaCat cells treated with MGO.

[0043] Figure 6 Figure showing the effect of the examples and comparative examples on the content of type I collagen (COL-1) in HaCat cells treated with MGO.

[0044] Figure 7 Figure showing the effect of the examples and comparative examples on the adhesion of HaCat cells treated with MGO.

[0045] Figure 8 Figure showing the effect of the examples and comparative examples on the activity of tyrosinase in mouse melanoma cells.

[0046] Figure 9 Figure showing the effect of the examples and comparative examples on the fluorescence intensity of β-galactosidase after UVB irradiation of zebrafish skin.

[0047] Figure 10 Figure showing the effect of the examples and comparative examples on the content of glycated hemoglobin after UVB irradiation of zebrafish skin.

[0048] Figure 11 Figure showing the effect of the examples and comparative examples on the fluorescence staining of β-galactosidase after UVB irradiation of zebrafish skin, where A and B are the normal and UVB model groups, C-H are Comparative Examples 1-6, and I-L are Examples 1-4.

[0049] Figure 12 Figure showing the effect of the examples and comparative examples on the reduction of fish tail fins after UVB irradiation of zebrafish skin, where A and B are the normal and UVB model groups, C-H are Comparative Examples 1-6, and I-L are Examples 1-4. DETAILED DESCRIPTION

[0050] The concept and the technical effects of the present application will be described clearly and completely in combination with the embodiments, so that the purpose, features and effects of the present application can be fully understood. Obviously, the described embodiments are only some of the embodiments of the present application, but not all the embodiments. Based on the embodiments of the present application, other embodiments obtained by those skilled in the art without creative labor are within the protection scope of the present application.

[0051] In the present embodiment, curcumin is selected from Solabio Biotech Co., Ltd. (content ≥98%); the extracts of Alpiniae oxyphyllae and Alpiniae galangae are self-extracted by a designed method: 200 g of dried rhizome of Alpiniae oxyphyllae and 200 g of dried fruit of Alpiniae galangae (produced in Bozhou, Anhui) are crushed into powder by a pulverizer, and then ethanol is added to obtain the extracts of the two medicinal materials respectively by ultrasonic, and different concentrations of ethanol eluate of the extracts of the two medicinal materials are obtained by using D101 macroporous resin, and the 60%-90% ethanol eluate is dried to obtain extract, which is stored.

[0052] The relevant commercially available goods can be selected by those skilled in the art according to actual needs. Unless otherwise specified, the reagents and materials used in the following embodiments are commercially available.

[0053] Example 1: A composition with anti-aging effect

[0054] An anti-aging composition, which is composed of the following components in parts by weight: curcumin 2 parts, extract of Alpiniae oxyphyllae 2.4 parts, extract of Alpiniae galangae 3.6 parts.

[0055] Example 2: A composition with anti-aging effect

[0056] An anti-aging composition, which is composed of the following components in parts by weight: curcumin 2 parts, extract of Alpiniae oxyphyllae 3 parts, extract of Alpiniae galangae 3 parts.

[0057] Example 3: A composition with anti-aging effect

[0058] An anti-aging composition, which is composed of the following components in parts by weight: curcumin 2 parts, extract of Alpiniae oxyphyllae 3.6 parts, extract of Alpiniae galangae 2.4 parts

[0059] Example 4: A composition with anti-aging effect

[0060] An anti-aging composition, which is composed of the following components in parts by weight: curcumin 2 parts, extract of Alpiniae oxyphyllae 4.2 parts, extract of Alpiniae galangae 1.8 parts.

[0061] Comparative Example 1: A composition with anti-aging effect

[0062] An anti-aging composition, which is composed of the following components in parts by weight: curcumin 8 parts.

[0063] Comparative Example 2 A composition having an anti-aging effect

[0064] An anti-aging composition consisting of the following components by weight: 8 parts of Alpinia officinarum extract.

[0065] Comparative Example 3 A composition having an anti-aging effect

[0066] An anti-aging composition consisting of the following components by weight: 8 parts of Alpinia galanga extract.

[0067] Comparative Example 4 A composition having an anti-aging effect

[0068] An anti-aging composition consisting of the following components by weight: 2 parts of curcumin, 6 parts of Alpinia officinarum extract.

[0069] Comparative Example 5 A composition having an anti-aging effect

[0070] An anti-aging composition consisting of the following components by weight: 2 parts of curcumin, 6 parts of Alpinia galanga extract.

[0071] Comparative Example 6 A composition having an anti-aging effect

[0072] An anti-aging composition consisting of the following components by weight: 4 parts of Alpinia officinarum extract, 4 parts of Alpinia galanga extract.

[0073] I. Effect of the anti-aging composition on the survival rate of MGO-treated HaCat cells.

[0074] 1. Materials and methods

[0075] Composition preparation: curcumin, Alpinia officinarum extract and Alpinia galanga extract were dissolved in DMSO, mixed and added to the culture medium, so that the final concentration of the examples and comparative examples was 10 μg / mL. Human immortalized keratinocytes (HaCat cells, Shanghai Cell Bank of the Chinese Academy of Sciences), fetal bovine serum, DMEM medium containing double antibodies, 0.25% trypsin (all from Gibico, USA), 37°C constant temperature incubator, 96-well plate (USA Sumefei), microplate reader (Switzerland Tecan).

[0076] Experimental procedure

[0077] HaCat cells were inoculated into complete culture medium and cultured at 37°C, 5% CO2. The cultured HaCat cells were digested with trypsin containing 0.25% EDTA, and 10 4The cells were inoculated in 96-well plates. When the cells in the 96-well plates were incubated to about 50% per well, the culture medium containing the compositions of Comparative Examples 1-6 and Example 1-4 was added for pre-treatment, and the blank group and the model group were added with an equal amount of fresh culture medium. After 12 h, the culture medium was aspirated, and the model group, the comparative example group and the example group were added with culture medium containing 1000 μM MGO, and the blank group was added with an equal amount of fresh culture medium, with a final volume of 100 μL per well. After 24 h of continuous culture, 20 μL of MTT was added per well, and incubated in a 37°C incubator for 4 h. The supernatant was aspirated, 150 μL of DMSO was added per well, and it was allowed to stand for 5 min. The absorbance value was immediately determined at a wavelength of 570 nm on a microplate reader.

[0078] Cell viability (%) = OD value of drug-treated group / OD of solvent control group x 100%.

[0079] 2. Results

[0080] Table 1 Effect of different compositions on cell viability values

[0081] Group Cell viability (%) Blank group 100.64±1.69 Model group 50.88±0.82## Comparative Example 1 63.96±1.29* Comparative Example 2 58.51±0.13* Comparative Example 3 61.84±1.38* Comparative Example 4 67.65±3.49* Comparative Example 5 61.28±2.10* Comparative Example 6 65.36±0.66** Example 1 79.15±2.56** Example 2 75.86±1.67** Example 3 88.49±3.05** Example 4 79.62±1.58**

[0082] *P < 0.05, **P < 0.01 compared with the model group, ##P < 0.01 compared with the blank group.

[0083] As shown in Table 1, the precursor of glycosylated end products, methylglyoxal (MGO) 1000 μM, had an induced damage effect on HaCat cells, with a survival rate of about 50%. In the comparative examples, curcumin, high galangal extract and cardamom extract, and the combination of two of them, had a significant protective effect on MGO-induced cell death. However, after mixing the three in different proportions in the examples, the cell survival rate increased again, indicating that the combination of the three had a synergistic protective effect.

[0084] II. Effect of the anti-aging composition on MGO-treated HaCat cells at the gene and protein levels

[0085] 1. Materials and methods

[0086] Composition preparation: curcumin, alpiniae oxyphyllae fructus extract and myristica fragrans extract were dissolved in DMSO, mixed and then added to the culture medium to make the final concentration of examples and comparative examples 10 μg / mL. Human immortalized keratinocytes (HaCat cells, Shanghai Cell Bank of Chinese Academy of Sciences), fetal bovine serum (Gibico, USA), DMEM medium containing double antibodies (Gibico, USA), 0.25% trypsin (Gibico, USA), 37°C constant temperature incubator (Binder, Germany), six-well plate (USA), RNA extraction kit (Nanjing Novozyme), reverse transcription kit (Nanjing Novozyme), DEPC water (Nanjing Novozyme), qPCR primer (Nanjing Novozyme), human type I collagen Elisa kit (Shanghai Bohu Biological), Light Cycler 480 real-time fluorescence quantitative PCR instrument (Roche, USA).

[0087] Experimental steps

[0088] The cultured HaCat cells were digested with trypsin containing 0.25% EDTA and inoculated on a six-well plate at 1.5×10 5 cells per well. When the cells in the six-well plate were incubated to 60%-70% per well, the culture medium containing the compositions of comparative examples 1-6 and examples 1-4 was added for pretreatment, and the blank group and the model group were added with an equal amount of fresh culture medium. After 12 h, the culture medium was aspirated, and the model group, the comparative example group and the example group were added with culture medium containing 1000 μM MGO, and the blank group was added with an equal amount of fresh culture medium, and the final volume of all wells was 2 mL. After 24 h of continuous culture, the culture medium was aspirated, and the RNA of each group of cells was extracted according to the operation of the RNA extraction kit, and the cell RNA was reverse transcribed into cDNA by the cell RNA reverse transcription kit, and finally the expression amount was determined by primers and fluorescent reagents on a real-time fluorescence quantitative PCR instrument.

[0089] The cells were cultured again as above, and the cells of all groups were collected and operated according to the steps of the human type I collagen Elisa kit instruction manual to determine the results.

[0090] 2. Results

[0091] 2.1 Inflammatory factors

[0092] Table 2 Effect of different compositions on expression of IL-6 and IL-8

[0093]

[0094]

[0095] *P<0.05, **P<0.01, ***P<0.001 compared with the model group, and ####P<0.0001 compared with the blank group.

[0096] As shown in Table 2, MGO has a significant pro-inflammatory effect on HaCat cells, and the induced inflammatory factors IL-6 and IL-8 are significantly increased in gene expression. The use of the comparative example has a significant down-regulating effect on the increase of IL-6 and IL-8 genes, and the use of the three examples has a more obvious down-regulating effect, indicating that they have a synergistic anti-inflammatory effect.

[0097] 2.2 MMP-9

[0098] Table 3 Effect of different compositions on MMP-9 gene expression

[0099] Group Gene expression amount Blank group 1.01±0.01 Model group 2.93±0.05### Comparative Example 1 1.76±0.05** Comparative Example 2 1.89±0.02** Comparative Example 3 1.81±0.08** Comparative Example 4 1.79±0.16* Comparative Example 5 1.80±0.07** Comparative Example 6 1.71±0.05** Example 1 1.01±0.01** Example 2 2.93±0.05** Example 3 1.76±0.05** Example 4 1.89±0.02**

[0100] *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 compared with the model group, ##P < 0.01 compared with the blank group.

[0101] During the glycosylation process, overexpression of MMP-9 can destroy the extracellular matrix, weaken the adhesion and migration function of skin cells, and is closely related to the formation of wrinkles due to impaired skin barrier.

[0102] As shown in Table 3, MGO induction can greatly increase the expression of MMP-9 gene in HaCat cells, and the comparative example has a down-regulating effect. Compared with the comparative example, the examples basically have a synergistic effect in terms of down-regulating MMP-9.

[0103] 2.3 AGER

[0104] Table 4 Effect of different compositions on AGER gene expression

[0105] Group Gene expression amount Blank group 1.08±0.08 Model group 2.17±0.01## Comparative Example 1 1.34±0.05** Comparative Example 2 1.54±0.04** Comparative Example 3 1.63±0.02** Comparative Example 4 1.30±0.02** Comparative Example 5 1.31±0.10* Comparative Example 6 1.32±0.03** Example 1 1.06±0.05** Example 2 0.84±0.02*** Example 3 0.79±0.01**** Example 4 1.14±0.01****

[0106] *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 compared with the model group, ##P < 0.01 compared with the blank group. At the cellular level, the AGER gene encodes a glycosylation end product receptor (RAGE), which mediates the binding of glycosylation end products (AGEs) and their ligands as a signal transduction receptor, and high expression can cause skin aging and inflammation.

[0107] As shown in Table 4, MGO induction can cause overexpression of AGER gene in HaCat cells, and the comparative example has a significant down-regulating effect on this overexpression. The examples further prove that the use of the three examples has a stronger down-regulating effect, and has a synergistic effect.

[0108] 2.4. Type I collagen (COL-1) expression

[0109] Table 5 Effect of different compositions on type I collagen expression

[0110] Group COL-1 (μg / L) Blank group 3.68±0.02 Model group 2.31±0.01### Comparative Example 1 3.08±0.03** Comparative Example 2 2.98±0.03** Comparative Example 3 2.48±0.01** Comparative Example 4 3.08±0.01*** Comparative Example 5 3.11±0.05** Comparative Example 6 2.79±0.06* Example 1 3.18±0.02*** Example 2 4.03±0.01**** Example 3 4.79±0.01**** Example 4 4.38±0.11**

[0111] *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 vs model group, ###P<0.001 vs blank group.

[0112] COL-1 collagen is an important component of the extracellular matrix of the skin, and plays an important role in the migration and adhesion of skin cells. It is also an important reason for the damage of the aging barrier of the skin and the generation of wrinkles.

[0113] As shown in Table 5, MGO induction can cause low expression of COL-1 protein in HaCat cells. The treatment of the comparative group has a significant up-regulation effect on this low expression, and the example group proves that the three combined up-regulation is stronger, even promotes the generation and expression of COL-1, showing a synergistic effect.

[0114] III. Effect of the Anti-aging Composition on the Adhesion Function of MGO-treated HaCat Cells

[0115] 1. Materials and Methods

[0116] Composition preparation: curcumin, high-quality ginger extract and cardamom extract were dissolved in DMSO, mixed and then added to the culture medium, so that the final concentration of the example and comparative example was 10 μg / mL. Human immortalized keratinocytes (HaCat cells, Shanghai Cell Bank of Chinese Academy of Sciences), fetal bovine serum (Gibico, USA), DMEM culture medium containing double antibodies (Gibico, USA), 0.25% trypsin (Gibico, USA), 37°C constant temperature incubator (Binder, Germany), six-well plates (Sumefly, USA), 96-well plates (Sumefly, USA) and CCK-8 kit (Solebao, China).

[0117] Experimental steps

[0118] The cultured HaCat cells were digested with trypsin containing 0.25% EDTA, and 1.5×10 5The cells were inoculated in a six-well plate. When the cells in the six-well plate were incubated to 60%-70% per well, the culture medium containing the compositions of Comparative Examples 1-6 and Example 1-4 was added for pretreatment, and the blank group and the model group were added with an equal amount of fresh culture medium. After 12 h, the culture medium was aspirated, and the model group, the comparative example group and the example group were added with culture medium containing 1000 μM MGO, and the blank group was added with an equal amount of fresh culture medium, and the final volume of all the holes was 2 mL. After 24 h of continuous culture, the culture medium was aspirated, the residual culture medium was washed away with PBS per well, and resuspended by trypsin digestion, and then inoculated into a 96-well plate, 6 holes per group, and continued to culture for 4 h. CCK-8 working solution was directly added to the first 3 holes of each group, and the culture medium was poured out from the last 3 holes, and then washed 3 times with PBS to remove the unadherent cells, and then CCK-8 working solution was added, and then incubated at 37°C for 1.5 h, and then the absorbance value at 450 nm wavelength was detected by an enzyme-labeled instrument, and then the percentage of adherent cells was calculated.

[0119] Adhesion rate: Adhesion Rate (%) = the adhesive cells / the total cells x 100%.

[0120] 1. Results

[0121] Table 6 Influence of different compositions on cell adhesion ability

[0122] Group Adhesion rate (%) Blank group 77.97±8.07 Model group 28.23±11.01### Comparative Example 1 64.91±1.47*** Comparative Example 2 53.91±4.35** Comparative Example 3 65.50±2.0*** Comparative Example 4 62.80±2.24** Comparative Example 5 67.35±8.79** Comparative Example 6 58.30±3.77** Example 1 74.03±2.46*** Example 2 78.07±8.91*** Example 3 86.13±6.07*** Example 4 80.14±11.61***

[0123] **P<0.01, ***P<0.001 compared with the model group, and ###P<0.001 compared with the blank group.

[0124] As shown in Table 6, the induction of MGO can reduce the adhesion ability of HaCat cells, the treatment of the comparative example can increase the adhesion ability of the cells, and the example proves that the up-regulation of the combination of the three is stronger, and has a synergistic effect.

[0125] IV. Influence of the anti-aging composition on the activity of tyrosinase in mouse melanoma cells

[0126] 1. Materials and methods

[0127] Composition preparation: curcumin, high-lingyangjiang extract and cardamom extract were dissolved in DMSO, mixed and then added into the culture medium, so that the final concentration of the example and the comparative example was 10 μg / mL. Mouse melanoma cells (B 16 F 10 Cells, Shanghai Cell Library of Chinese Academy of Sciences), fetal bovine serum (Gibico, USA), DMEM culture medium containing double antibodies (Gibico, USA), trypsin containing 0.25% EDTA (Gibico, USA), 37°C constant temperature incubator (Binder, Germany) and 96-well plate (Thermo, USA).

[0128] Experimental procedure

[0129] Mouse melanoma cells (B 16 F 10 ) were inoculated into complete medium and cultured at 37°C in 5% CO2. The cultured B 16 F 10 cells were digested with trypsin containing 0.25% EDTA and inoculated into 96-well plates at 5 x 10 3 cells per well. When the cells in the 96-well plates were incubated to 50% per well, the medium containing the compositions of Comparative Examples 1-6 and Examples 1-4 was added, and the same amount of fresh medium was added to the blank group. After 24 h of culture, the cells were washed twice with PBS, 50 μL / mL of 1% Triton X-100 solution was added, and the cells were rapidly frozen at -20°C for 1 h. After the cells were thawed to room temperature, 10 μL of 0.1% L-dopa solution was added, and the reaction was performed in a 37°C constant temperature water bath for 4 h. The absorbance A value was measured at 492 nm. The relative activity of tyrosinase was calculated.

[0130] Relative activity of tyrosinase (%) = A 492 of the administration group / A 492 of the blank group x 100%.

[0131] 2. Results

[0132] Table 7 Effect of different compositions on the activity of tyrosinase

[0133]

[0134]

[0135] *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 compared with the blank group.

[0136] Tyrosinase is a key enzyme in the synthesis of melanin in organisms, and it is of great significance to find effective enzyme inhibitors from natural plant extracts. Using mouse melanoma cells as a whitening model, it can be seen from Table 7 that the activity of the enzyme after treatment with the blank group and the comparative example group decreased significantly, and the enzyme inhibition of the example group had a synergistic effect compared with the comparative example group.

[0137] V. Effects of the Anti-aging Composition on Glycation Aging-related Factors Induced by UVB Irradiation in Zebrafish Models Zebrafish models are widely used in the field of cosmetics. Zebrafish larvae within 5 days after fertilization (5dpf) do not need to eat and are not considered animals in terms of ethics, so they are used in cosmetic skin replacement experiments. They can be used not only for raw material screening and safety evaluation but also for efficacy evaluation of final products.

[0138] 1. Materials and Methods

[0139] Composition Preparation: Curcumin, Alpiniae oxyphyllae Fructus extract, and Myristica fragrans extract were dissolved in DMSO, mixed, and then added to the culture solution to make the final concentration of the examples and comparative examples 1.25 μg / mL. Wild-type AB strain zebrafish, ultraviolet phototherapy instrument (SH4B, Shanghai Sigma High Technology Co., Ltd.), electric focusing continuous zoom fluorescence microscope, microplate reader, six-well plate, glycosylated hemoglobin ELISA detection kit (Shanghai Lianzu Biology), and cell senescence β-galactosidase staining kit (China Biyun Tian Biology).

[0140] Experimental Steps

[0141] 2dpf wild-type AB strain zebrafish, 30 fish per group, 3 mL in a 6-well plate, before UVB irradiation, the culture solution containing the corresponding content of the anti-aging composition of the example group and the comparative group was added to the 6-well plate. The total administration time was maintained for 48 h. The endpoint of the experiment was 4dpf, and the zebrafish in each group were collected, photographed, and the fluorescence intensity was calculated using Image J software. The index was determined according to the kit instructions.

[0142] 2.1 Experimental Results

[0143] Table 8 Effects of Different Compositions on the Fluorescence Intensity of β-galactosidase in the Epidermis of Zebrafish

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[0146] ****P <0.0001 compared with the model group, and ####P <0.0001 compared with the blank group.

[0147] Cell senescence refers to the gradual decline in cell proliferation and differentiation ability and physiological function during the performance of life activities over time. When cells senesce, the activity of senescence-associated β-galactosidase (SA-βgal) increases significantly, so the activity of SA-βgal is often detected to distinguish normal cells from senescent cells. Currently, SA-βgal detection is often used as one of the markers of cell senescence after cells are treated with different conditions or compounds.

[0148] From Table 8, after the zebrafish was irradiated by UVB and fluorescently dyed, the fluorescence intensity of the model group was greatly improved compared with the blank group, while the fluorescence intensity of the comparative example group and the example group was significantly reduced compared with the model group, and the example group showed synergistic down-regulation. 2.2

[0150] Table 9 Effect of different compositions on glycated hemoglobin concentration of zebrafish

[0151] Group Glycated hemoglobin concentration (ng / mL) Blank group 44.04±0.53 Model group 64.37±0.55### Comparative Example 1 56.37±1.58* Comparative Example 2 56.94±0.89** Comparative Example 3 56.54±1.67* Comparative Example 4 55.23±2.46* Comparative Example 5 56.20±2.26* Comparative Example 6 56.82±1.85* Example 1 51.62±0.91** Example 2 51.89±2.26** Example 3 51.84±1.16** Example 4 54.62±1.05*

[0152] *P < 0.05, **P < 0.01, compared with the model group, ###P < 0.001 compared with the blank group.

[0153] Clinical studies have found that the glycated hemoglobin content in the body of normal aging population is significantly higher than that of normal young people. The Maillard reaction, sugar autoxidation and other metabolic pathways (such as glycolysis) produce dicarbonyl compounds, which produce AGEs, and combine with hemoglobin in the body to form glycated hemoglobin, which accumulates continuously with age, that is, serum AGEs is linearly related to aging, indicating that glycated hemoglobin is one of the important indicators for evaluating aging.

[0154] From Table 9, after the zebrafish was irradiated by UVB, the glycated hemoglobin content of the model group was increased compared with the blank group, while the content of the comparative example and the example group was significantly reduced compared with the model group, indicating that the anti-aging composition can reduce the increase of glycated hemoglobin induced by UVB, and has synergistic down-regulation.

[0155] The above is a further description of the present application combined with specific examples, but these examples are only exemplary and do not constitute any limitation on the scope of the present application. Those skilled in the art should understand that the details and forms of the technical solutions of the present application can be modified or replaced without departing from the spirit and scope of the present application, and such modifications and replacements all fall within the protection scope of the present application.

[0156] In all the following pictures, Control represents the blank group, MGO represents the MGO-induced model group, UVB represents the UVB-induced model group, A-F represent Comparative Examples 1-6, and 1-4 represent Examples 1-4, respectively.

Claims

1. An anti-aging composition characterized in that, In mass parts, it consists of 0.1-8 parts of curcumin, 0.1-8 parts of high pungent ginger extract and 0.1-5 parts of extract of cardamom; the high pungent ginger extract and the cardamom extract are obtained by the following steps: taking 200 g of high pungent ginger dried rhizome and 200 g of cardamom dried fruit as raw medicinal materials, crushing them into powder by a pulverizer, adding ethanol to obtain the extract of each by ultrasonic, using D101 macroporous resin to obtain different concentrations of ethanol eluate of the extract of each, taking the 60%-90% ethanol eluate part to dry, obtaining the extract, and storing. ​ 2. The composition of claim 1, wherein: In mass parts, it consists of 0.1-4 parts of curcumin, 1-5 parts of high pungent ginger extract and 1-4 parts of extract of cardamom.

3. Use of the composition according to claim 1 or 2 in the preparation of a cosmetic product.

4. Use according to claim 3, characterized in that: The cosmetic product includes cleansing milk, cosmetic water, emulsion, facial mask, cream and serum.

5. A cosmetic product, characterized by, The cosmetic product contains the composition according to claim 1 or 2 and cosmetically acceptable adjuvant.

6. The cosmetic product according to claim 5, characterized in that, The adjuvant contains at least one of emollients, emulsifiers, thickening agents, humectants, pH regulators, skin protectants and preservatives.

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