Soothing, anti-wrinkle and anti-oxidation energy skin care composition

Abstract

The invention discloses a soothing, anti-wrinkle and anti-oxidation energy skin care composition, the composition comprises dendrobe, cordyceps sinensis, sophora flower and ribose, and the dendrobe, the cordyceps sinensis and the sophora flower comprise aqueous extracts and / or alcohol extracts of the dendrobe, the cordyceps sinensis and the sophora flower. The energy skin care composition has multiple effects of relieving, wrinkle resistance, oxidation resistance and the like, and based on contrast experiments, the four components can achieve synergistic interaction, so that a better effect is achieved, and a new thought is provided for development of related cosmetics.

Description

Technical Field

[0001] The invention relates to the technical field of cosmetics, and in particular to an energy skin care composition that is soothing, anti-wrinkle and anti-oxidant. Background Art

[0002] The skin is the largest organ of the human body and is also the part of the human body that is most vulnerable to damage and pollution. Therefore, skin care is extremely important for maintaining skin health. In addition to pathological factors such as skin diseases and skin injuries, factors that endanger skin health also include physiological skin aging (also known as skin aging). Pathological factors can be treated with medical means, but physiological skin aging can only be delayed or improved with functional substances. With the improvement of material conditions and economic conditions, consumers are paying more and more attention to the problem of skin aging and are looking for products that can effectively improve or alleviate skin aging.

[0003] Skin aging is a complex process related to changes in the ultrastructure and biochemistry of the skin. These changes can be the result of endogenous aging (i.e. aging that occurs with age) and / or exogenous aging, either alone or in combination. For example, photoaging caused by UV radiation will further deepen the endogenous aging process; lifestyle, environment, disease, and gravity will also have an impact on skin aging. As researchers delve deeper into the mechanism of skin aging, principles such as how physiological and metabolic changes caused by age lead to changes in skin structure and how external stimuli cause skin damage are constantly being revealed, so that cosmetics with relevant functions can be designed in a targeted manner. However, despite this, relatively few active substances with anti-aging, soothing and other functions have been found in the prior art, and many substances are still facing multiple tests such as actual use effects and safety. Therefore, there are still only a handful of products that can actually produce relevant functions to achieve skin care.

[0004] Therefore, there is an urgent need to develop an energy skin care composition that can achieve soothing, anti-wrinkle and anti-oxidation effects, thereby providing new ideas for the development of cosmetics with related effects. Summary of the invention

[0005] The present invention aims to solve at least one of the above-mentioned technical problems existing in the prior art. To this end, the purpose of the present invention is to provide a soothing, anti-wrinkle and anti-oxidation energy skin care composition. The present invention is based on four raw materials, namely, dendrobium, cordyceps, sophora japonica and ribose, to develop a composition with multiple functions such as soothing, anti-wrinkle and anti-oxidation, and based on comparative experiments, it is found that the four ingredients can synergize with each other to achieve better effects.

[0006] The first aspect of the present invention provides a composition comprising dendrobium, cordyceps sinensis, sophora japonica flower and ribose.

[0007] In some embodiments of the present invention, the dendrobium, cordyceps and sophora japonica include water extracts and / or alcohol extracts of dendrobium, cordyceps and sophora japonica.

[0008] In the present invention, the terms "water extract" and "alcohol extract" refer to extracts obtained by extracting the whole plant, medicinal part or edible part using water or alcohol solution as a solvent according to a conventional extraction method.

[0009] In some embodiments of the present invention, the dendrobium includes Dendrobium nobile.

[0010] In some embodiments of the present invention, the dendrobium, cordyceps sinensis and sophora japonica are water extracts of dendrobium, cordyceps sinensis and sophora japonica.

[0011] In the present invention, Dendrobium nobile Lindl is the stem and leaf parts of Dendrobium nobile Lindl, a plant of the Orchidaceae family and the genus Dendrobium.

[0012] In the present invention, Cordyceps sinensis (Ophiocordyceps sinensis (Berk.) GH Sung, JM Sung, Hywel-Jones & Spatafora) is a fruiting body of Cordyceps sinensis of the family Claviceps and the genus Nematodes.

[0013] In the present invention, the Sophora japonica Linn flower is a flower of the Fabaceae family, the Sophora japonica Linn genus.

[0014] In some embodiments of the present invention, in the composition, the mass ratio of Dendrobium officinale, Cordyceps sinensis and Sophora japonica flower is 5-15:0.2-1:0.2-2.

[0015] In some embodiments of the present invention, in the composition, the mass ratio of Dendrobium officinale, Cordyceps sinensis and Sophora japonica flower is 8-12:0.5-1:0.5-1.5.

[0016] In some embodiments of the present invention, the composition further comprises an adjuvant.

[0017] In some embodiments of the present invention, the adjuvant includes a pharmaceutically acceptable adjuvant, a cosmetically acceptable adjuvant or a food acceptable adjuvant.

[0018] In some embodiments of the present invention, the pharmaceutically acceptable adjuvant, cosmetically acceptable adjuvant or food acceptable adjuvant includes: solvents and propellants: such as ethanol, propylene glycol, glycerol, etc.; solubilizers and cosolvents: such as polysorbates, polyoxyethylene fatty acid esters, benzoic acid, potassium iodide, etc.; emulsifiers: such as Tween 80, Span 80, etc.; colorants: such as food coloring, natural coloring; binders: such as starch slurry, sodium carboxymethyl cellulose (CMC-Na), hydroxypropyl methylcellulose (HPMC), etc.; disintegrants: such as dry starch, sodium carboxymethyl starch (CMS-Na); fillers: such as starch, sucrose, lactose, etc.; lubricants: such as magnesium stearate, micro powder silica gel; Wetting agents: such as polysorbates; osmotic pressure regulators: such as sodium chloride, glucose; stabilizers: such as ascorbic acid, vitamin E; flow aids: such as silica gel; flavoring agents: including sweeteners (such as sucrose, saccharin sodium) and fragrances (such as peppermint oil, lemon oil); preservatives: such as benzoic acid, benzalkonium bromide, etc.; coating materials: such as sugar coating, film coating, etc.; release retardants: such as hydroxypropyl methylcellulose (HPMC), etc.; surfactants: such as Tween 80, Span 80; foaming agents and defoaming agents: such as sodium bicarbonate, etc.; thickeners: such as xanthan gum, carrageenan, etc.; inclusion agents: β-cyclodextrin, etc.; humectants: such as glycerol, propylene glycol; absorbents: such as activated carbon, silica gel, etc.; diluents: such as water, ethanol, etc.

[0019] Of course, those skilled in the art may also reasonably add other adjuvants based on actual use needs, including but not limited to the above-mentioned adjuvants.

[0020] In some embodiments of the present invention, the adjuvant comprises butanediol.

[0021] In some embodiments of the present invention, the composition comprises: by mass, 5-15 parts of dendrobium, 0.2-1 parts of cordyceps, 0.2-2 parts of sophora japonica, 40-50 parts of butanediol and 0.1-1 parts of ribose.

[0022] In some embodiments of the present invention, the composition comprises: by mass, 8-12 parts of dendrobium, 0.5-1 parts of cordyceps, 0.5-1.5 parts of sophora japonica, 42-45 parts of butanediol and 0.5-1 parts of ribose.

[0023] The second aspect of the present invention provides a method for preparing the composition described in the above aspect, comprising the following steps:

[0024] The extracts of dendrobium, cordyceps and sophora japonica are mixed and decolorized, and auxiliary agents and ribose are added to obtain the product.

[0025] or

[0026] The product is obtained by mixing dendrobium, cordyceps and sophora japonica flowers, extracting and decolorizing them, and then adding auxiliary agents and ribose.

[0027] In some embodiments of the present invention, the extract comprises a water extract and / or an alcohol extract.

[0028] In some embodiments of the present invention, the extract is an aqueous extract.

[0029] In some embodiments of the present invention, the adjuvant includes a pharmaceutically acceptable adjuvant, a cosmetically acceptable adjuvant or a food acceptable adjuvant.

[0030] In some embodiments of the present invention, the adjuvant comprises butanediol.

[0031] In some embodiments of the present invention, the preparation method is specifically as follows: grind dendrobium, cordyceps sinensis, and sophora japonica flowers, mix them, add water for water extraction, the water extraction temperature is 75-85°C, centrifuge, take the supernatant, concentrate under constant temperature and reduced pressure, decolorize, and then add butanediol and ribose to obtain.

[0032] In some embodiments of the present invention, the water extraction time is 1-3 hours.

[0033] In some embodiments of the present invention, the water extraction time is 2-3 hours.

[0034] In some embodiments of the present invention, the temperature of the isothermal reduced pressure concentration is 70-80°C.

[0035] In some embodiments of the present invention, the temperature of the isothermal reduced pressure concentration is 75°C.

[0036] In some embodiments of the present invention, the stopping condition of the isothermal reduced pressure concentration is: the specific gravity of the substance is 1.15-1.20.

[0037] Of course, those skilled in the art can also directly use commercially available extracts, mix them directly, and then decolorize them according to subsequent steps, add butanediol and ribose, etc. to prepare the above composition.

[0038] The third aspect of the present invention provides use of the composition described in the above aspects in the preparation of cosmetics or medicines.

[0039] In some embodiments of the present invention, the cosmetic or medicine has at least one of the following functions:

[0040] (1) Antioxidant;

[0041] (2) Anti-wrinkle;

[0042] (3) Soothing and anti-inflammatory.

[0043] In some embodiments of the present invention, the cosmetics are in the form of creams, frosts, honeys, milks, liquids, powders, tablets, gels, aerosols, aerosols, etc.

[0044] The fourth aspect of the present invention provides a product, which comprises the composition described in the above aspects and at least one excipient and / or carrier.

[0045] In some embodiments of the invention, the product is a cosmetic or a pharmaceutical.

[0046] In the present invention, the term "excipient" refers to an additive other than the main drug in a pharmaceutical preparation, such as an auxiliary agent that performs at least one function of bonding, filling, disintegration, lubrication, etc.

[0047] In the present invention, the term "carrier" refers to a substance used to carry and protect active ingredients in cosmetics or medicines, and its functions include: improving the stability and safety of the active ingredients, enhancing their skin permeability, etc.

[0048] The beneficial effects of the present invention are:

[0049] The present invention has developed a safe and effective natural material-based composition with multiple functions such as soothing, anti-wrinkle and anti-oxidation. The four raw materials of dendrobium, cordyceps, sophora japonica and ribose are combined to effectively improve the efficacy of the composition compared with a single substance. At the same time, the development of the composition of the present invention also provides a new idea for soothing, anti-wrinkle and anti-oxidation cosmetics, which has important practical and economic value. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] Figure 1 This is a graph showing the activity of macrophages treated with the composition of an example of the present invention.

[0051] Figure 2 The results of TNF-α content detection in different experimental groups.

[0052] Figure 3 This is a graph showing the activity of fibroblasts treated with the composition of an example of the present invention.

[0053] Figure 4 The figure shows the morphology of fibroblasts treated with the composition in the example of the present invention.

[0054] Figure 5 Fluorescence images of different experimental groups.

[0055] Figure 6 The results of ROS content detection in different experimental groups.

[0056] Figure 7 The results of type I collagen content detection in different experimental groups.

[0057] Figure 8The results of ATP content detection in different experimental groups.

[0058] Fig. 9 Ultrasound images of collagen density in different experimental groups.

[0059] Fig.10 Results of collagen density test in different experimental groups

[0060] Fig.11 This is the ATP content detection result of the synergistic enhancement experiment.

[0061] Fig.12 This is the result of TNF-α content detection in the synergistic enhancement experiment.

[0062] Fig.13 These are the collagen density test results for the synergistic enhancement experiment.

[0063] Fig.14 These are the results of type I collagen content testing in the synergistic enhancement experiment.

[0064] Fig.15 This is the ROS content detection result of the synergistic enhancement experiment. DETAILED DESCRIPTION

[0065] The present invention is further described in detail below by specific examples. Unless otherwise specified, the raw materials, reagents or devices used in the examples and comparative examples can be obtained from conventional commercial sources or can be obtained by prior art methods. Unless otherwise specified, the experiments or test methods are conventional methods in the art.

[0066] In the following examples, the ribose used specifically refers to D-ribose with CAS: 50-69-1. Of course, those skilled in the art may also use ribose obtained from other sources or prepared by conventional methods in the art, including but not limited to D-ribose, and the present invention is not intended to limit the selection of ribose.

[0067] In the following examples, the Dendrobium nobile extract, Cordyceps sinensis extract, and Sophora japonica flower extract may be commercially available products, or may be extracted using conventional methods in the art.

[0068] In the following examples, the extract of Dendrobium nobile, the extract of Cordyceps sinensis, and the extract of Sophora japonica are all water extracts, and the extraction method is as follows:

[0069] After the raw materials are crushed, water is added, and then they are heated and extracted at 70-90°C, and then centrifuged (or filtered) to obtain the product.

[0070] In the following examples, the Dendrobium nobile extract, Cordyceps sinensis extract, and Sophora japonica flower extract can be extracted separately and then mixed to prepare the composition, or the raw materials can be mixed and extracted uniformly to prepare the composition.

[0071] Example 1 A soothing, anti-wrinkle and anti-oxidant energy skin care composition

[0072] In this embodiment, the raw materials of the soothing, anti-wrinkle and anti-oxidant energy skin care composition are: by mass, 10 parts of Dendrobium nobile, 0.5 parts of Cordyceps sinensis, 1 part of Sophora japonica, 45 parts of butylene glycol, and 0.5 parts of ribose.

[0073] The preparation method is:

[0074] After the Dendrobium nobile, Cordyceps sinensis and Sophora japonica flowers are crushed separately, they are mixed according to the above mass parts, and then 43 mass parts of water are added, and extracted at 80°C for 2.5h. Centrifuge and take the supernatant. The supernatant is concentrated under reduced pressure at 75°C until the specific gravity of the substance is 1.15-1.20 under the heating condition, and then it is decolorized (200-300 mesh coconut shell activated carbon decolorization). According to the above mass parts, butanediol and ribose are added to the decolorized solution, and the mixture is fully mixed and filtered to obtain the product.

[0075] During large-scale production, the process also includes: letting the filtrate obtained by filtration stand, conducting microbiological tests, and then filling and shipping the product after it passes the tests.

[0076] Example 2 A soothing, anti-wrinkle and anti-oxidant energy skin care composition

[0077] In this embodiment, the raw materials of the soothing, anti-wrinkle and anti-oxidant energy skin care composition are: by mass, 12 parts of Dendrobium nobile, 1 part of Cordyceps sinensis, 0.5 parts of Sophora japonica, 42.5 parts of butylene glycol, and 1 part of ribose.

[0078] The preparation method is the same as that of Example 1. The amount of water added is maintained at 43 parts by mass.

[0079] Example 3 A soothing, anti-wrinkle and anti-oxidant energy skin care composition

[0080] In this embodiment, the raw materials of the soothing, anti-wrinkle and anti-oxidant energy skin care composition are: by mass, 8 parts of Dendrobium nobile, 0.5 parts of Cordyceps sinensis, 1.5 parts of Sophora japonica, 46 parts of butylene glycol, and 1 part of ribose.

[0081] The preparation method is the same as that of Example 1. The amount of water added is maintained at 43 parts by mass.

[0082] Example 4 Effect Verification of Composition

[0083] (1) Verification of the soothing effect of the composition:

[0084] In this embodiment, the cytotoxicity of the composition is determined based on the cytotoxicity test of macrophages, and the soothing effect of the composition is detected based on the in vitro experiment of detecting the content of inflammatory factors (TNF-α) after stimulation. The specific detection method is as follows:

[0085] A. Cytotoxicity:

[0086] After the macrophages were revived, they were cultured routinely. When the plating rate reached about 60%, the cells were transferred to a 96-well plate and incubated overnight in a CO2 incubator (37°C, 5% CO2). As shown in Table 1, the composition in Example 1 was diluted to different concentrations with culture medium (DMEM).

[0087] Table 1 Composition test concentration

[0088]

[0089] When the cell plating rate in the 96-well plate reached 40%-60%, the grouping and administration were carried out (3 parallels per group). Among them, 200 μL of DMEM culture solution containing the corresponding concentration of the composition was added to each well of the experimental group; the zeroing group had no cell inoculation and only 200 μL of DMEM culture solution was added; the blank group had cells inoculated and only 200 μL of DMEM culture solution was added. After the administration was completed, the 96-well plate was placed in a CO2 incubator and incubated for 24 hours (37°C, 5% CO2).

[0090] After the cells were incubated for 24 hours, the supernatant was discarded, and MTT working solution (0.5 mg / mL) was added, and the cells were incubated at 37°C in the dark for 4 hours. After the incubation, the supernatant was discarded, 150 μL of DMSO was added to each well, and the OD value was read at 490 nm. The relative cell viability was calculated according to the following formula:

[0091]

[0092] The results are shown in Table 2 and Figure 1 shown.

[0093] Table 2MTT test results

[0094]

[0095] According to the MTT and morphological results, it can be considered that the composition of the present invention does not show obvious cytotoxicity within the concentration range of 0.15625% (v / v) based on the macrophage in vitro experimental results.

[0096] B. Soothing effect verification:

[0097] At the same time, the experiment was repeated according to the above method, and the dosing scheme was adjusted as follows: 1.8 mL of culture solution was added to each well of the blank control group and the negative control group, and 1.8 mL of culture solution containing the corresponding concentration of the composition was added to each well of the experimental group. After the dosing was completed, the 6-well plate was placed in a CO2 incubator for incubation for 2 hours (37°C, 5% CO2). Three parallels were set for each group.

[0098] After the incubation, 200 μL of PBS containing 1 μg / mL LPS was added to each well of the remaining groups except the blank control group, and the cells were placed in a CO2 incubator for further 22 h. After the incubation, the cell culture supernatant was collected by centrifugation, and the TNF-α content was detected using a commercially available ELISA kit. At the same time, the inhibition rate was calculated according to the following formula.

[0099]

[0100] GraphPad Prism was used to draw the graphs, and the results were expressed as Mean ± SD. The t-test was used for statistical analysis between the groups. All statistical analyses were two-tailed. P < 0.05 was considered to be significantly different, and P < 0.01 was considered to be extremely significantly different.

[0101] The results are as follows Figure 2 And as shown in Table 3.

[0102] Table 3 TNF-α content detection results

[0103] Group Average concentration (pg / mL) SD P-value Inhibition rate (vs NC) BC 851.38 17.41 / / NC 51108.45 3949.29 0.000## / 0.15626% (v / v) composition 43229.05 1708.57 0.034 15.42％

[0104] Among them, when the t-test method was used for statistical analysis, compared with the BC group, the significance was indicated by #, P<0.05 was indicated by #, and P<0.01 was indicated by ##; compared with the NC group, the significance was indicated by *, P<0.05 was indicated by *, and P<0.01 was indicated by **.

[0105] The results showed that compared with the BC group, the TNF-α content of the NC group increased significantly, indicating that the LPS stimulation conditions were effective. Compared with the NC group, the TNF-α content of the combination also decreased significantly, and the inhibition rate could reach 15.42%.

[0106] In summary, based on the macrophage experimental results, compared with the control group, the content of inflammatory factor (TNF-α) in the composition at a concentration of 0.15625% (vw) was significantly decreased, and the inhibition rate was 15.42%, indicating that the sample at this concentration can inhibit the secretion of inflammatory factor (TNF-α) and has a soothing effect.

[0107] (2) Antioxidant effect verification of the composition:

[0108] In this embodiment, the cytotoxicity of the composition is determined based on a cytotoxicity test of fibroblasts, and the antioxidant effect of the composition is detected based on an in vitro experiment to detect the change in ROS content after UVA irradiation. The specific detection method is:

[0109] A. Cytotoxicity and morphological changes:

[0110] The fibroblast toxicity test was performed according to the method in (1). The test concentration of the composition was the same as that in Table 1. At the same time, a 24-well plate was used to select the concentrations shown in Group 1 to Group 5 for repeated experiments (3 parallels, each adding 1 mL of the culture medium of the composition of the corresponding concentration), and the supernatant was discarded after incubation for 24 hours, and observation and photography were performed using an inverted microscope.

[0111] The results are shown in Table 4 and Figure 3-4 shown.

[0112] Table 4 MTT test results

[0113]

[0114] According to the MTT and morphological results, it can be considered that the composition of the present invention does not show obvious cytotoxicity within the concentration range of 0.3125% (v / v) based on the in vitro experimental results of fibroblasts.

[0115] B. Antioxidant effect verification based on fibroblasts:

[0116] Fibroblasts were cultured in 6-well plates, and then different dosing was performed according to the groups. 2 mL of culture medium without additives was added to each well of the negative control group (NC) and the blank control group (BC). 2 mL of culture medium containing the corresponding concentration (0.3125%) of the composition was added to each well of the experimental group. After the dosing was completed, it was placed in a CO2 incubator and incubated for 24 hours (37°C, 5% CO2). Three parallels were set up for each group.

[0117] After the incubation, all groups except the blank control group were irradiated with UVA at a dose of 30 J / cm 2 . After irradiation, continue incubation for 24 hours. After incubation, wash the cells 3 times with PBS, add 1 mL of 10 μM DCFH-DA probe (Biyuntian) to each well, and incubate in a CO2 incubator for 30 minutes (37°C, 5% CO2). Discard the culture medium containing DCFH-DA, wash 3 times with PBS, digest the cells with trypsin (0.25%), wash the cells once with PBS, add fresh PBS, and use flow cytometry to detect the ROS content therein, and calculate the downregulation rate according to the following formula:

[0118]

[0119] GraphPad Prism was used to draw the graphs, and the results were expressed as Mean ± SD. The t-test was used for statistical analysis between the groups. All statistical analyses were two-tailed. P < 0.05 was considered to be significantly different, and P < 0.01 was considered to be extremely significantly different.

[0120] The results are as follows Figure 5 And as shown in Table 5.

[0121] Table 5 ROS content detection and analysis results

[0122]

[0123]

[0124] Wherein, MFI represents mean fluorescence intensity. When the t-test method was used for statistical analysis, the significance was indicated by #, P<0.05 was indicated by #, and P<0.01 was indicated by ## compared with the BC group; the significance was indicated by *, P<0.05 was indicated by *, and P<0.01 was indicated by ** compared with the NC group.

[0125] The results showed that the ROS content of the NC group increased significantly compared with the BC group, indicating that the UVA irradiation stimulation conditions were effective. Compared with the NC group, the ROS content of the combination decreased significantly, and its downregulation rate reached 20.86%.

[0126] Based on fibroblasts, compared with the control group, the reactive oxygen species (ROS) content of the composition at a concentration of 0.3125% was significantly reduced, with a downregulation rate of 20.86%, indicating that the composition, at least at this concentration, can reduce the reactive oxygen species (ROS) content and has an antioxidant effect.

[0127] (3) Verification of the anti-wrinkle effect of the composition:

[0128] In this embodiment, the anti-wrinkle efficacy of the composition to be tested is evaluated based on the changes in the content of type I collagen (Collagen I) and adenosine triphosphate (ATP) in fibroblasts after UVA irradiation, and the changes in the collagen density of ex vivo skin tissue after UVA and UVB combined irradiation. The specific detection method is:

[0129] A. Anti-wrinkle effect verification based on fibroblasts:

[0130] Based on the verified cytotoxicity and morphological conditions in (2), a composition with a relevant concentration gradient was selected for anti-wrinkle experiment, and the specific steps were as follows:

[0131] Fibroblasts were cultured in 6-well plates, and then different dosing was performed according to the groups. 2 mL of culture medium without additives was added to each well of the negative control group (NC) and the blank control group (BC). 2 mL of culture medium containing the corresponding concentration (0.3125%) of the composition was added to each well of the experimental group. After the dosing was completed, it was placed in a CO2 incubator and incubated for 24 hours (37°C, 5% CO2). Three parallels were set up for each group.

[0132] After the incubation, all groups except the blank control group were irradiated with UVA at a dose of 30 J / cm 2 . After irradiation, continue incubation for 24 hours. After incubation, collect part of the cell culture, centrifuge and take the supernatant, and use a commercially available ELISA detection kit to detect the type I collagen content. In addition, after washing the 6-well plate with 2 mL / well PBS, add 200 μL / well cell lysis solution, blow and lyse the cells, and use a commercially available ATP detection kit (Biyuntian) to detect the ATP content.

[0133] Based on the test results, the improvement rate is calculated according to the following formula:

[0134]

[0135] GraphPad Prism was used to draw the graphs, and the results were expressed as Mean ± SD. The t-test was used for statistical analysis between the groups. All statistical analyses were two-tailed. P < 0.05 was considered to be significantly different, and P < 0.01 was considered to be extremely significantly different.

[0136] The results are as follows Figure 6-7 As shown in Table 6-7.

[0137] Table 6 Type I collagen content detection results

[0138] Group Average concentration (ng / mL) SD P-value Improvement rate (vs NC) BC 62.75 1.84 / / NC 39.97 1.26 0.000## / 0.3125% (v / v) composition 54.75 1.40 0.000** 36.98％

[0139] Among them, when the t-test method was used for statistical analysis, compared with the BC group, the significance was indicated by #, P<0.05 was indicated by #, and P<0.01 was indicated by ##; compared with the NC group, the significance was indicated by *, P<0.05 was indicated by *, and P<0.01 was indicated by **.

[0140] The results showed that compared with the BC group, the type I collagen content of the NC group decreased significantly, indicating that the stimulation conditions were effective. Compared with the NC group, the type I collagen content after the composition treatment increased significantly, with an increase rate of 36.98%.

[0141] Table 7 ATP content test results

[0142] Group ATP content (μM / mg) SD P-value Improvement rate (vs NC) BC 587.90 33.12 / / NC 323.41 6.77 0.000## / 0.3125% (v / v) composition 383.22 15.87 0.004** 18.49％

[0143] Among them, when the t-test method was used for statistical analysis, compared with the BC group, the significance was indicated by #, P<0.05 was indicated by #, and P<0.01 was indicated by ##; compared with the NC group, the significance was indicated by *, P<0.05 was indicated by *, and P<0.01 was indicated by **.

[0144] The results showed that compared with the BC group, the ATP content in the NC group decreased significantly, indicating that the stimulation conditions were effective. Compared with the NC group, the ATP content after the combination treatment increased significantly, with an increase rate of 18.49%.

[0145] B. Anti-wrinkle effect verification based on ex vivo skin tissue:

[0146] Take freshly obtained skin tissue and immerse it in 75% alcohol, wash it for 30 seconds, and then wash it three times with sterile PBS buffer. Cut the washed skin tissue into 24±2mm 2 The small pieces were placed in a culture mold with the epidermis facing upward and the dermis facing downward. The culture mold was then transferred to a 6-well plate. 3.7 L of in vitro tissue culture medium (Huibai Biotechnology) was added to each well. The plates were incubated at 37°C and 5% CO2, and the medium was changed every day.

[0147] After 2 days of in vitro skin tissue culture, the cells were irradiated and divided into groups for drug administration (three parallel groups per group). The irradiation dose of the negative control group (NC) and the experimental group was UVA (30 J / cm 2 ) and UVB (50J / cm 2 ), irradiated continuously for 4 days. After each irradiation, fresh culture medium was replaced and drug treatment was carried out. The experimental group adopted the method of subsurface drug administration. The negative control group added 1mL of culture medium without additives, while the experimental group added 1mL of culture medium containing the composition of the corresponding concentration (0.3125%) to each well. At the same time, a blank control group (BC) was set up without any treatment or drug administration. After 4 days of irradiation and drug administration, culture was continued for 3 days and drug administration was continued.

[0148] After the experiment, the collagen density was measured using a skin ultrasound machine, and the improvement rate was calculated using the following formula:

[0149]

[0150] GraphPad Prism was used to draw the graphs, and the results were expressed as Mean ± SD. The t-test was used for statistical analysis between the groups. All statistical analyses were two-tailed. P < 0.05 was considered to be significantly different, and P < 0.01 was considered to be extremely significantly different.

[0151] The results are as follows Figure 8-9 and as shown in Table 8.

[0152] Table 8 Collagen density test results

[0153] Group Average density (%) SD P-value Improvement rate (vs NC) BC 13.67 2.52 / / NC 6.33 0.58 0.008## / 0.3125% (v / v) composition 10.00 1.00 0.005** 57.98％

[0154] Among them, when the t-test method was used for statistical analysis, compared with the BC group, the significance was indicated by #, P<0.05 was indicated by #, and P<0.01 was indicated by ##; compared with the NC group, the significance was indicated by *, P<0.05 was indicated by *, and P<0.01 was indicated by **.

[0155] As shown in the figure, in the ultrasound image of the dermis, both green (low-density area) and yellow-white (high-density area) represent collagen. Therefore, the greater the proportion of the relevant color in the image and the brighter it is, the higher the collagen density in the corresponding tissue. The results showed that compared with the BC group, the collagen density of the NC group decreased significantly, indicating that the stimulation conditions were effective. Compared with the NC group, the collagen density after treatment with the combination increased significantly, with an increase rate of 57.98%.

[0156] In summary, combined with the results of fibroblasts and ex vivo skin tissue, it can be found that compared with the control group, the composition of the present invention can significantly increase the content of type I collagen (Collagen I) and adenosine triphosphate (ATP) at a concentration of 0.3125% (vw), with an increase rate of 36.98% and 18.49% respectively. For ex vivo skin tissue, it can significantly increase the collagen density, with an increase rate of 57.98%. This shows that the composition of the present invention has an anti-wrinkle effect.

[0157] Example 5: Synergistic Effect Verification

[0158] In order to fully illustrate the synergistic effect between the components in the composition of the embodiment of the present invention, the inventors conducted the following verification experiments.

[0159] According to the test methods in the above examples, the soothing, anti-wrinkle and antioxidant effects of the single components (water extracts of Dendrobium nobile, Cordyceps sinensis and Sophora japonica treated according to the examples, and nucleic acids) and the composition in Example 1 were verified. The dosage concentration of each experimental group was 0.3125% (v / v), and the others were the same as in Example 4.

[0160] The results are as follows Figure 10-14 and as shown in the table below.

[0161] Table 9 ATP content test results

[0162]

[0163]

[0164] Table 10 TNF-α content detection results

[0165]

[0166] Table 11 Collagen density test results

[0167]

[0168] Table 12 Type I collagen content detection results

[0169]

[0170]

[0171] Table 13 ROS content detection results

[0172] Sample Group Test results SD P-value Downward adjustment rate Blank control (BC) 81521.6 1164.50 / / Normal control (NC) 134545.0 2493.79 0.0003### / Sophora japonica flower extract 118399.6 3143.18 0.0134* 12.0％ Dendrobium nobile extract 121090.5 1723.54 0.0437* 10.0％ Cordyceps sinensis extract 127817.8 1540.80 0.5000 5.0％ Ribose 115708.7 11314.48 0.0040** 14.0％ Composition 106475.8 11044.93 ＜0.0001**** 20.9％

[0173] It can be seen from the results that, compared with a single component, the composition of the present invention has a more significant effect on increasing ATP content, collagen density and type I collagen content, and can significantly reduce TNF-α content and ROS content. Therefore, it can be determined that the components in the present invention can produce a significant synergistic effect.

[0174] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A composition, characterized in that The composition comprises dendrobium, cordyceps sinensis, sophora japonica and ribose; Wherein, the dendrobium, cordyceps and sophora japonica include water extract and / or alcohol extract of dendrobium, cordyceps and sophora japonica; Preferably, the dendrobium includes Dendrobium nobile.

2. The composition according to claim 1, characterized in that In the composition, the mass ratio of dendrobium, cordyceps and sophora japonica is 5-15:0.2-1:0.2-2; Preferably, in the composition, the mass ratio of dendrobium, cordyceps and sophora japonica is 8-12: 0.5-1：0.5-1.5。 3. The composition according to claim 1 or 2, characterized in that The composition also includes an adjuvant; The adjuvant includes a pharmaceutically acceptable adjuvant, a cosmetically acceptable adjuvant or a food acceptable adjuvant; Preferably, the adjuvant comprises butanediol.

4. The composition according to claim 3, characterized in that The composition comprises: by weight, 5-15 parts of dendrobium, 0.2-1 parts of cordyceps sinensis, 0.2-2 parts of sophora japonica, 40-50 parts of butanediol and 0.1-1 parts of ribose; Preferably, the composition comprises: by mass, 8-12 parts of dendrobium, 0.5-1 parts of cordyceps, 0.5-1.5 parts of sophora japonica, 42-45 parts of butanediol and 0.5-1 parts of ribose.

5. A method for preparing the composition according to any one of claims 1 to 4, comprising the steps of: The extracts of dendrobium, cordyceps and sophora japonica are mixed and decolorized, and then auxiliary agents and ribose are added to obtain the product; or The product is obtained by mixing dendrobium, cordyceps and sophora japonica flowers, extracting and decolorizing them, and then adding auxiliary agents and ribose.

6. The preparation method according to claim 5, characterized in that: The extract includes water extract and / or alcohol extract.

7. The preparation method according to claim 5, characterized in that: The adjuvant includes a pharmaceutically acceptable adjuvant, a cosmetically acceptable adjuvant or a food acceptable adjuvant; Preferably, the adjuvant comprises butanediol.

8. Use of the composition according to any one of claims 1 to 4 in the preparation of cosmetics or medicines.

9. The use according to claim 8, characterized in that The cosmetic or medicine has at least one of the following functions: (1) Antioxidant; (2) Anti-wrinkle; (3) Soothing and anti-inflammatory.

10. A product, characterized in that The product comprises the composition according to any one of claims 1 to 4, and at least one excipient and / or carrier; Preferably, the product is a cosmetic or a medicine.

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