A composition for resisting post-inflammatory hyperpigmentation of the skin, its preparation method and application

Abstract

The application discloses a kind of compositions to resist skin inflammation post pigmentation, the preparation raw material of the composition includes white atractylodes and lophatherum gracile, and the mass ratio of the white atractylodes and lophatherum gracile is (1-50):(6-15).The present application can regulate micro ribonucleic acid miR-146a expression, control skin inflammation from source, reduce inflammatory factor release, regulate the content of melanin, maintain the healthy homeostasis of epidermis, achieve the effect of whitening and brightening skin color, fading color spots, and restoring the healthy state of skin.And resist the negative effect of inflammation on skin color, by adjusting the expression of key inflammatory factors PGE2 affecting skin pigmentation in inflammation and melanin formation, so as to resist inflammatory pigmentation.

Description

Technical Field

[0001] This invention relates to a composition, preparation method and application for resisting post-inflammatory hyperpigmentation of the skin, and relates to A61K, specifically to the field of cosmetic products. Background Technology

[0002] As people place increasing importance on their appearance, the pursuit of skin whitening and fading dark spots is becoming more and more fervent. Plant-based whitening methods offer relatively gentle effects and high safety. Researchers have found that sun exposure and ultraviolet light are not the only factors causing skin darkening. Skin can also experience pigmentation after inflammation, mechanical damage, trauma, and phototherapy, especially in people of color, where pigment is more easily deposited, leading to a darker complexion. Therefore, addressing post-inflammatory pigmentation is a crucial approach to solving skin whitening problems. This invention focuses on the role of inflammatory regulatory factors in the entire pigmentation process, restoring the skin to its normal state from the source by inhibiting skin inflammation and regulating microRNA expression, thereby reducing skin pigmentation caused by various factors.

[0003] Chinese invention patent CN201210110090.0 discloses a herbal whitening and freckle-removing facial mask. It involves scientifically blending various Chinese herbs, pulverizing them into an extremely fine powder, and then mixing them with fresh milk to form a paste. This achieves the effects of nourishing and beautifying the skin, moisturizing and preventing wrinkles, lightening melanin, and whitening and removing freckles. However, it uses a large number of Chinese herbs, requires immediate preparation, has a complex preparation method, is inconvenient to use, and is costly. Chinese invention patent CN201810386075.6 discloses a cosmetic composition containing Chinese herbal extracts and its preparation method. It uses multiple Chinese herbal skincare products in synergy, providing comprehensive care and conditioning through inhibiting melanin production, moisturizing, and acne removal, achieving whitening, freckle removal, moisturizing, anti-wrinkle, anti-inflammatory, and acne-removing effects, thus achieving both symptomatic and root-cause treatment. However, it involves extracting the Chinese herbs separately and then combining the extracts for use, resulting in a less significant whitening effect and failing to specifically address post-inflammatory hyperpigmentation. Summary of the Invention

[0004] In order to specifically address the problem of post-inflammatory hyperpigmentation and achieve the effects of reducing melanin content, anti-inflammatory, antioxidant, and repairing the skin barrier, the first aspect of the present invention provides a composition for resisting post-inflammatory hyperpigmentation of the skin. The raw materials for preparing the composition include Atractylodes macrocephala and Lophatherum gracile, wherein the mass ratio of Atractylodes macrocephala to Lophatherum gracile is (1-50):(6-15).

[0005] In a preferred embodiment, the mass ratio of Atractylodes macrocephala to Lophatherum gracile is (5-20):(9-12).

[0006] In a preferred embodiment, the mass ratio of Atractylodes macrocephala to Lophatherum gracile is 5:12 or 20:9.

[0007] In a preferred embodiment, the raw materials for preparing the composition also include one or more of the following: a complex enzyme, a monohydric alcohol solution, a pH adjuster, a polyol, and a solvent.

[0008] In a preferred embodiment, the monohydric alcohol solution is an aqueous ethanol solution.

[0009] In a preferred embodiment, the polyol is selected from one or a combination of several of propylene glycol, butanediol, glycerol, sorbitol, and ethylene glycol.

[0010] In a preferred embodiment, the enzyme activity of the composite enzyme is 35,000-50,000 g. -1 The amount of compound enzyme added is 0.5-3% of the substrate mass.

[0011] In a preferred embodiment, the enzyme activity of the composite enzyme is 40,000-45,000 g. -1 The amount of compound enzyme added is 1-2% of the amount of substrate to be hydrolyzed.

[0012] In a preferred embodiment, the enzyme activity of the composite enzyme is 40,000 g. -1 The amount of compound enzyme added is 1% of the mass of the substrate to be hydrolyzed.

[0013] In a preferred embodiment, the complex enzyme is selected from one or a combination of cellulase, pectinase, protease, and amylase.

[0014] In a preferred embodiment, the composite enzyme is a combination of cellulase and pectinase. Preferably, the weight ratio of cellulase to pectinase is (1-3):1, and more preferably, the weight ratio of cellulase to pectinase is 2:1.

[0015] During the experiment, the applicant discovered that extracting Atractylodes macrocephala and Lophatherum gracile at a weight ratio of 20:9, compared to extracting Atractylodes macrocephala and Lophatherum gracile separately, resulted in a higher tyrosinase inhibition rate. The applicant speculates that this may be because, after enzymatic hydrolysis with a compound enzyme, components in Lophatherum gracile can promote the dissolution of flavonoids in Atractylodes macrocephala into the solvent, improving the extraction efficiency of flavonoids. Simultaneously, at the 20:9 weight ratio, the extraction rate of atractylodes macrocephala's trace component, atractylodes lactone, is higher, which can synergistically work with the Lophatherum gracile extract to regulate the expression of microRNA miR-146a, inhibiting melanin deposition caused by skin inflammation at its source. The applicant further discovered that atractylodes lactone, atractylodes glycoside, and atractylodes aglycone in Atractylodes macrocephala can synergistically work with flavonoid glycosides in Lophatherum gracile to regulate the expression of PGE2, a key inflammatory factor affecting skin pigmentation in inflammation and melanin formation, reducing the release of inflammatory factors. When Atractylodes macrocephala and Lophatherum gracile are extracted together, the extract contains higher levels of trace components such as atractylodes lactone, atractylodes glycoside, and atractylodes aglycone, which in turn enhance the ability to inhibit tyrosinase activity.

[0016] A second aspect of the present invention provides a method for preparing a composition for resisting post-inflammatory hyperpigmentation of the skin, comprising the following steps:

[0017] S1 Weigh Atractylodes macrocephala and Lophatherum gracile according to the weight ratio, crush them and add them to the solvent, adjust the pH with a pH adjuster, and heat the reaction.

[0018] Add the compound enzyme to S2 and stir well;

[0019] S3 enzyme hydrolysis for 1-2 hours, followed by high-temperature inactivation of the complex enzyme;

[0020] S4 ultrasonic extraction for 20-60 min, followed by centrifugation, filtration, and collection of extract and residue;

[0021] S5 uses a monohydric alcohol solution for ultrasonic extraction of the filter residue, filters it, and collects the filtrate;

[0022] S6 concentrates the filtrate, removes the monohydric alcohol solution, and obtains the extract;

[0023] S7 dissolves the extract in a polyol, then mixes it with the extract solution, and finally filters it through an ultrafiltration membrane to obtain the final product.

[0024] In a preferred embodiment, the pH value in step S1 is 5-6, the heating temperature is 40-55℃, and the weight ratio of the total mass of Atractylodes macrocephala and Lophatherum gracile to the solvent is 1:(5-15).

[0025] In a preferred embodiment, the pH value in step S1 is 5, the heating temperature is 50°C, and the total mass ratio of Atractylodes macrocephala and Lophatherum gracile to the solvent is 1:10.

[0026] In a preferred embodiment, the ultrasonic extraction power in step S4 is 140-300W, and the ultrasonic temperature is 40-55℃.

[0027] In a preferred embodiment, the ultrasonic extraction power in step S4 is 200-280W, and the ultrasonic temperature is 45-20℃.

[0028] In a preferred embodiment, the ultrasonic extraction power in step S4 is 260W and the ultrasonic temperature is 50℃.

[0029] In a preferred embodiment, the ultrasonic extraction time in step S4 is 30-40 min; the ultrasonic extraction time in step S5 is 20-30 min; more preferably, the ultrasonic extraction time in step S4 is 35 min; and the ultrasonic extraction time in step S5 is 25 min.

[0030] In a preferred embodiment, the temperature for high-temperature inactivation of the complex enzyme in step S3 is 80-95℃.

[0031] In a preferred embodiment, the filtration in steps S4 and S5 is microfiltration, and the pore size of the microfiltration membrane is 0.1-1 μm; the pore size of the ultrafiltration membrane in step S7 is 0.01-0.05 μm.

[0032] In a preferred embodiment, the microfiltration membrane has a pore size of 0.45 μm; and the ultrafiltration membrane in step S7 has a pore size of 0.01-0.05 μm.

[0033] In a preferred embodiment, the concentration method in step S6 is water bath heating or rotary evaporation concentration.

[0034] In a preferred embodiment, the volume concentration of the monohydric alcohol solution in step S5 is 60-90%.

[0035] In a preferred embodiment, the monohydric alcohol solution in step S5 is an aqueous ethanol solution with a volume concentration of 70%.

[0036] In a preferred embodiment, the volume ratio of the extract dissolved in polyol to the extract is 1:(1-3); preferably, the volume ratio of the extract dissolved in polyol to the extract is 1:(1-2); more preferably, the volume ratio of the extract dissolved in polyol to the extract is 1:1.

[0037] A third aspect of the invention provides the application of a composition for resisting post-inflammatory hyperpigmentation of the skin in cosmetics or skin care products.

[0038] In a preferred embodiment, the solvents in which the prepared composition is soluble when applied include, but are not limited to, deionized water, ethanol, propylene glycol, and butanediol.

[0039] In one preferred embodiment, the skincare products include, but are not limited to, toners, lotions, sprays, creams, masks, and serums. The cosmetics include, but are not limited to, foundations, BB creams, and cushion foundations.

[0040] Compared with the prior art, the present invention has the following beneficial effects:

[0041] (1) The composition for resisting post-inflammatory hyperpigmentation of the skin described in this invention, wherein Atractylodes macrocephala and Lophatherum gracile are mixed and extracted in a weight ratio of 20:9 or 5:12, has a higher tyrosinase inhibition rate after use compared with the individual extraction of Atractylodes macrocephala and Lophatherum gracile.

[0042] (2) The composition for resisting post-inflammatory hyperpigmentation of the skin described in this invention is extracted by a combination of cellulase and pectinase. The composition has a high content of the trace component atractylodes lactone, which can regulate the expression of microRNA miR-146a, regulate skin inflammation from the source, reduce the release of inflammatory factors, regulate the content of melanin, maintain the healthy homeostasis of the epidermis, and achieve the effects of whitening and brightening skin tone, fading spots, and restoring the skin to a healthy state.

[0043] (3) The composition of the present invention for resisting post-inflammatory hyperpigmentation of the skin resists the negative effects of inflammation on skin color by regulating the expression of PGE2, a key inflammatory factor that affects skin pigmentation in inflammation and melanin formation, thereby resisting inflammatory hyperpigmentation. Attached Figure Description

[0044] Figure 1 These are data graphs showing the inhibition rates of the compositions in Examples 1-9 and Comparative Examples 1-2 against the inflammatory factor PGE2;

[0045] Figure 2 The graphs show the miR-146a expression levels of the compositions in the blank control group, irradiation group, Examples 5 and 7, and Comparative Examples 1 and 2.

[0046] Figure 3 These are experimental graphs showing the effects of the compositions of the blank control group, irradiation group, Examples 5 and 7, and Comparative Examples 1 and 2 on melanin expression levels.

[0047] Figure 4 Before (top) and after four weeks (bottom) using an emulsion containing 1 wt%, 2 wt%, and 5 wt% of the composition of Example 5 (from left to right), the skin brightening is improved. Detailed Implementation

[0048] Example 1

[0049] A composition for resisting post-inflammatory hyperpigmentation of the skin, the raw materials for preparing the composition include Atractylodes macrocephala and Lophatherum gracile, wherein the mass ratio of Atractylodes macrocephala and Lophatherum gracile is 1:6.

[0050] The raw materials for preparing the composition also include one or more of the following: a complex enzyme, a monohydric alcohol solution, a pH adjuster, a polyol, and a solvent.

[0051] The monohydric alcohol solution is an aqueous ethanol solution with a volume concentration of 70%. The pH adjuster is an acetate-ammonium acetate buffer solution.

[0052] The polyol is butanediol. The solvent is deionized water.

[0053] The enzyme activity of the complex enzyme is 40,000 g. -1 The amount of the compound enzyme added is 1% of the weight of the substrate. The compound enzyme is a combination of cellulase and pectinase in a weight ratio of 2:1.

[0054] A method for preparing a composition for resisting post-inflammatory hyperpigmentation of the skin, comprising the following steps:

[0055] S1 Weigh Atractylodes macrocephala and Lophatherum gracile according to the weight ratio, crush them and add them to the solvent, adjust the pH with a pH adjuster, and heat the reaction.

[0056] Add the compound enzyme to S2 and stir well;

[0057] S3 enzyme hydrolysis for 1 hour, followed by high-temperature inactivation of the complex enzyme;

[0058] S4 ultrasonic extraction for 35 min, followed by centrifugation, filtration, and collection of extract and residue;

[0059] S5 uses a monohydric alcohol solution for ultrasonic extraction of the filter residue, filters it, and collects the filtrate;

[0060] S6 concentrates the filtrate, removes the monohydric alcohol solution, and obtains the extract;

[0061] S7 dissolves the extract in a polyol, then mixes it with the extract solution, and finally filters it through an ultrafiltration membrane to obtain the final product.

[0062] In step S1, the pH value is 5, the heating temperature is 50℃, and the total mass of Atractylodes macrocephala and Lophatherum gracile is 1:10 with the weight of the solvent.

[0063] In step S5, the weight ratio of the monohydric alcohol solution to the filter residue is 20:1.

[0064] In step S4, the ultrasonic extraction power is 260W and the ultrasonic temperature is 50℃. In step S5, the ultrasonic extraction time is 25min.

[0065] The temperature at which the complex enzyme is inactivated at high temperature in step S3 is 85°C.

[0066] The microfiltration membrane has a pore size of 0.45 μm; the ultrafiltration membrane in step S7 has a pore size of 0.01-0.05 μm.

[0067] In step S6, the concentration method is rotary evaporation. In step S7, the weight ratio of polyol to extract is 59:1.

[0068] The volume ratio of the extract dissolved in polyol to the extract solution is 1:1.

[0069] Example 2

[0070] A composition for resisting post-inflammatory hyperpigmentation of the skin and its preparation method are described. The specific steps are the same as in Example 1, except that the mass ratio of Atractylodes macrocephala and Lophatherum gracile is 5:6.

[0071] Example 3

[0072] A composition for resisting post-inflammatory hyperpigmentation of the skin and its preparation method are described. The specific steps are the same as in Example 1, except that the mass ratio of Atractylodes macrocephala and Lophatherum gracile is 20:6.

[0073] Example 4

[0074] A composition for resisting post-inflammatory hyperpigmentation of the skin and its preparation method are described. The specific steps are the same as in Example 1, except that the mass ratio of Atractylodes macrocephala and Lophatherum gracile is 50:6.

[0075] Example 5

[0076] A composition for resisting post-inflammatory hyperpigmentation of the skin and its preparation method are described. The specific steps are the same as in Example 1, except that the mass ratio of Atractylodes macrocephala and Lophatherum gracile is 5:12.

[0077] Example 6

[0078] A composition for resisting post-inflammatory hyperpigmentation of the skin and its preparation method are described. The specific steps are the same as in Example 1, except that the mass ratio of Atractylodes macrocephala and Lophatherum gracile is 5:15.

[0079] Example 7

[0080] A composition for resisting post-inflammatory hyperpigmentation of the skin and its preparation method are described. The specific steps are the same as in Example 1, except that the mass ratio of Atractylodes macrocephala and Lophatherum gracile is 20:9.

[0081] Example 8

[0082] A composition for resisting post-inflammatory hyperpigmentation of the skin and its preparation method are described. The specific steps are the same as in Example 1, except that the mass ratio of Atractylodes macrocephala and Lophatherum gracile is 20:12.

[0083] Example 9

[0084] A composition for resisting post-inflammatory hyperpigmentation of the skin and its preparation method are described, with the specific steps being the same as in Example 1, except that the mass ratio of Atractylodes macrocephala and Lophatherum gracile is 20:15.

[0085] Comparative Example 1

[0086] A composition for resisting post-inflammatory hyperpigmentation of the skin and its preparation method are described. The specific steps are the same as in Example 1, except that the mass ratio of Atractylodes macrocephala and Lophatherum gracile is 1:0.

[0087] Comparative Example 2

[0088] A composition for resisting post-inflammatory hyperpigmentation of the skin and its preparation method are described. The specific steps are the same as in Example 1, except that the mass ratio of Atractylodes macrocephala and Lophatherum gracile is 0:1.

[0089] Performance testing

[0090] 1. Cytotoxicity assessment assay:

[0091] The MTT assay is a widely used experimental method for detecting cytotoxicity or cell proliferation by measuring the number of viable cells. Its principle is that succinate dehydrogenase in the mitochondria of viable cells reduces the water-soluble yellow salt MTT (3-(4,5-dimethylthiazol-2)-2,5-diphenyltetrazolium bromide) to a water-insoluble blue formazan, which is then deposited in the cells. Dead cells do not have this function. The resulting crystalline formazan is generally dissolved in DMSO (dimethyl sulfoxide) and the absorbance is measured. The specific experimental method involves preparing a 96-well plate at a density of 1 × 10⁻⁶ cells / well. 4 100 μL each of DMEM medium containing 10 wt% bovine serum and keratinocytes (HaCaT) were seeded at a density of cells / well and cultured for 24 hours. The culture was then replaced with serum-free medium. Different concentrations of the combined solution (50 μg / mL, 100 μg / mL, 200 μg / mL) were added to the serum-free medium for treatment, followed by 24 hours of culture. The medium was then removed, and the cells were treated with 20 μL of MTT solution (5 mg / mL) at 37°C for 2 hours. After removing the MTT solution, 200 μL of isopropanol was added to the cells, and the mixture was shaken for 30 min to completely dissolve the formazan crystals. The absorbance was measured at 570 nm, and cell viability was calculated using the following formula.

[0092]

[0093] The blank group was tested without the composition. Table 1 shows the cytotoxicity results of the compositions obtained in Examples 1-9 and Comparative Examples 1-2.

[0094] Table 1

[0095]

[0096]

[0097]

[0098] 2. Tyrosinase activity inhibition test:

[0099] In the biosynthesis of melanin in the skin, tyrosinase is a key enzyme that acts on dopa to form dopaquinone, which then spontaneously undergoes a series of reactions to ultimately form melanin. Tyrosinase catalyzes the conversion of dopa to dopaquinone in a phosphoric acid solution at pH 6.8, and the absorbance can be measured at 475 nm using a spectrophotometer. Raw materials that inhibit tyrosinase activity can reduce the conversion of dopa to dopaquinone, thereby lowering the absorbance. The inhibitory effect of the raw materials on tyrosinase activity can be evaluated based on changes in absorbance.

[0100] As shown in Table 2, phosphate buffer solution with pH 6.8 was added to the test tubes sequentially. Then, the anti-pigmentation compositions prepared in Examples 1-9 and Comparative Examples 1-2, along with arbutin, were added to each tube. Tyrosinase solution was then added to each tube, mixed, and incubated at 37°C for 10 minutes. Tyrosine solution was then added to each tube sequentially, and the tubes were incubated at 37°C for 5 minutes. The samples were then quickly transferred to cuvettes, and the absorbance was measured at 475 nm. The experimental groups were Examples 1-9 and Comparative Examples 1-2. The negative control was without the anti-pigmentation composition and arbutin, and the positive control was with arbutin added. The experimental groups, negative control, and positive control were zeroed using blank control 1, blank control 2, and blank control 3, respectively.

[0101] The concentration of the anti-pigmentation composition and arbutin solution used in the experiment was 1 mg / mL.

[0102] Tyrosinase inhibition rate = [(AB) / A] × 100%

[0103] A represents the absorbance value of the negative control, and B represents the absorbance value of the experimental group or the positive control.

[0104] The test results are shown in Table 3.

[0105] Table 2

[0106]

[0107]

[0108] Table 3

[0109] Tyrosinase inhibition rate (%) Tyrosinase inhibition rate (%) Positive control 59.32 Example 6 45.25 Example 1 31.75 Example 7 62.86 Example 2 36.72 Example 8 44.98 Example 3 42.17 Example 9 43.28 Example 4 44.78 Comparative Example 1 29.93 Example 5 60.24 Comparative Example 2 19.87

[0110] As shown in Table 3:

[0111] 1) The tyrosinase inhibition rate of the compositions in Examples 1-9 was higher than that of Comparative Examples 1-2, indicating that the extraction of two single raw materials by combining them in a certain proportion can achieve a better tyrosinase inhibition effect than the sum of the single extracts, with a technical effect of 1+1>2.

[0112] In Examples 1-9, the tyrosinase inhibition effect of the compositions increased with the increase of the proportion of Atractylodes macrocephala, reaching the highest point of tyrosinase inhibition rate after reaching a certain amount, then slowly decreasing, and then increasing again with the increase of the proportion of Lophatherum gracile, before leveling off at a certain limit.

[0113] 3. Inhibition of the inflammatory factor PGE2:

[0114] Cell culture: Raw264.7 cells in logarithmic growth phase were harvested, the culture medium was discarded, and the cells were washed twice with PBS (phosphate-buffered saline). 0.25 wt% trypsin was added, and the cells were incubated at 37°C for 5 min to stop digestion. The cells were centrifuged at 1000 rpm for 3 min, resuspended in complete DMEM medium, and counted. The cell concentration was adjusted to 5 × 10⁶ cells / mL. 5 100 μL / mL was added to each well of a 96-well plate, and the cells adhered after 24 hours.

[0115] Drug treatment: After culturing cells for 24 h, add 100 μL of different concentrations (2×) of the test substance and 1 μg / mL of LPS to each well, mix well, and incubate for 24 h ± 1 h. Then collect the supernatant for ELISA detection.

[0116] PGE2 inhibition rate % = (OD450 LPS experimental group - OD450 control group) / OD450 control group × 100%.

[0117] Depend on Figure 1 It can be known that:

[0118] (1) The PGE2 inhibition rate of the compositions in Examples 1-9 was higher than that of Comparative Examples 1-2, indicating that the PGE2 inhibition rate can be improved and the anti-inflammatory effect of the compositions can be enhanced by combining two single raw materials in a certain proportion.

[0119] (2) The PGE2 inhibition rate of the compositions in Examples 1-4 first increased and then decreased with the increase of the proportion of Atractylodes macrocephala. Therefore, the PGE2 inhibition rate is relatively high when Atractylodes macrocephala accounts for a certain proportion of the composition.

[0120] (3) The PGE2 inhibition rate of the compositions in Examples 6, 8 and 9 increased slowly with the increase of the proportion of Lophatherum gracile, indicating that Lophatherum gracile has a significant anti-inflammatory effect.

[0121] Based on this, considering the various effects, cost and stability of the composition, the preferred embodiments are Examples 5 and 7 (Atractylodes macrocephala: Lophatherum gracile = 5:12, 20:9).

[0122] 4. miR-146a and melanin expression level assay

[0123] Normal human epidermal melanocytes (NHEMs) were seeded in 96-well cell culture plates and cultured until the cell density reached 80%. The cultured normal human epidermal melanocytes (NHEMs) were then divided into the following three groups according to the experiment. miR-146a expression assay:

[0124] (1) Blank control group, no treatment was given;

[0125] (2) 15J / cm 2 UVA + 30mJ / cm 2 On day 2, total RNA was extracted from normal human epidermal melanocytes (NHEMs) in the UVB irradiation group and blank control group, and the expression level of miR-146a was detected by real-time quantitative PCR.

[0126] (3) Experimental group: 0.1 wt% of Examples 5 and 7, and Comparative Examples 1 and 2 were added to normal human epidermal melanocytes (NHEMs) and subjected to 15 J / cm² treatment. 2 UVA + 30mJ / cm 2 After UVB irradiation, total RNA was extracted on day 2, and the expression level of miR-146a was detected by real-time quantitative PCR.

[0127] △CT experimental group = CT target gene - CT internal reference gene, △CT control group = CT target gene - CT internal reference gene, △△CT = △CT experimental group - △CT control group;

[0128] The relative expression level of the target gene = 2 －△△CT ;

[0129] Melanin expression inhibition assay:

[0130] Cells were treated according to the above cell culture and grouping methods. After treatment, the cells were washed three times with PBS. After cell digestion was terminated, cell counting was performed. Then, 0.2 mol / L NaOH was added to dissolve the cells in a metal bath at 80°C. The cells were then added to an ELISA plate, and the differences in melanin production in each experimental group were detected by ultraviolet spectrophotometry.

[0131] Melanin inhibition rate % = (OD450 irradiation group - OD450 experimental group) / OD450 irradiation group × 100. Figure 2 , Figure 3 It can be known that:

[0132] (1) The miR-146a quantification values ​​of the compositions in Examples 5 and 7 were higher than those in Comparative Examples 1-2, indicating that the expression level of miR-146a can be increased by combining the two single raw materials in a certain proportion.

[0133] (2) The melanin expression levels of the compositions in Examples 5 and 7 were lower than those in Comparative Examples 1-2, and showed an inverse inhibitory effect on the expression level of miR-146a, indicating that the compositions can regulate the expression level of melanin by regulating the expression of microRNA miR-146a.

[0134] 5. Comparison Test of Human Body Whiteness Improvement

[0135] Thirty healthy volunteers, aged 24-38 years, were selected. Emulsions containing the composition of Example 5 (1 wt%, 2 wt%, and 5 wt% of the composition of Example 5, respectively) were used; the emulsion formulations are shown in Table 4. The emulsions were applied twice daily, morning and evening, for four consecutive weeks. Skin whiteness was measured using a CM2500D Spectrometer (KM, Japan), and the results are as follows: Figure 4 As shown. Figure 4 Comparison chart showing the improvement in skin whiteness after using an emulsion containing the composition of Example 5.

[0136] Table 4

[0137]

Claims

1. A composition for combating post-inflammatory hyperpigmentation of the skin, characterized in that it comprises, The preparation raw materials of the composition include Atractylodes and Ginkgo biloba leaves, and the mass ratio of the Atractylodes and the Ginkgo biloba leaves is 5:12 or 20:9; The preparation method of the composition for resisting post-inflammatory hyperpigmentation of skin comprises the following steps: S1, the Atractylodes and the Ginkgo biloba leaves are weighed according to a mass ratio, are crushed, are added into deionized water, and pH is adjusted by using a pH adjuster, and then the temperature is increased for reaction; S2, a composite enzyme is added, and stirring is uniformly performed; the composite enzyme is a combination of cellulase and pectinase; S3, enzymolysis is performed for 1-2 hours, and then the composite enzyme is inactivated at high temperature; S4, ultrasonic extraction is performed for 20-60 minutes, then centrifugal separation is performed, filtration is performed, and the extract and the residue are collected; S5, the residue is subjected to ultrasonic extraction by using a monohydric alcohol solution, filtration is performed, and the filtrate is collected; the monohydric alcohol solution is an ethanol aqueous solution, and the volume concentration is 60%-90%; S6, the filtrate is concentrated, the monohydric alcohol solution is removed, and an extract is obtained; S7, the extract is dissolved by using a polyhydric alcohol, and then is mixed with the extract, and after mixing, filtration is performed by using an ultrafiltration membrane, and the composition is obtained.

2. The composition to combat post-inflammatory hyperpigmentation according to claim 1, characterized in that, The enzyme activity of the complex enzyme is 35,000 U / g -1 - 50,000 U / g -1 The added amount of the complex enzyme is 0.5-3% of the mass of the enzymolysis substrate.

3. A process for the preparation of a composition for combating post-inflammatory hyperpigmentation of the skin according to any one of claims 1 to 2, characterized in that, The preparation method of the composition for resisting post-inflammatory hyperpigmentation of skin comprises the following steps: S1, the Atractylodes and the Ginkgo biloba leaves are weighed according to a mass ratio, are crushed, are added into deionized water, and pH is adjusted by using a pH adjuster, and then the temperature is increased for reaction; S2, a composite enzyme is added, and stirring is uniformly performed; the composite enzyme is a combination of cellulase and pectinase; S3, enzymolysis is performed for 1-2 hours, and then the composite enzyme is inactivated at high temperature; S4, ultrasonic extraction is performed for 20-60 minutes, then centrifugal separation is performed, filtration is performed, and the extract and the residue are collected; S5, the residue is subjected to ultrasonic extraction by using a monohydric alcohol solution, filtration is performed, and the filtrate is collected; the monohydric alcohol solution is an ethanol aqueous solution, and the volume concentration is 60%-90%; S6, the filtrate is concentrated, the monohydric alcohol solution is removed, and an extract is obtained; S7, the extract is dissolved by using a polyhydric alcohol, and then is mixed with the extract, and after mixing, filtration is performed by using an ultrafiltration membrane, and the composition is obtained.

4. The method for preparing the composition for resisting post-inflammatory hyperpigmentation of the skin according to claim 3, characterized in that, In the step S1, the pH value is 5-6, the temperature is increased to 40-55 DEG C, and the total mass of the Atractylodes and the Ginkgo biloba leaves is 1: (5-15) times the weight of the solvent.

5. The method of preparing a composition for combating post-inflammatory hyperpigmentation according to claim 3, characterized in that, In the step S4, the ultrasonic extraction power is 140-300 W, and the ultrasonic temperature is 40-55 DEG C.

6. The method of preparing a composition for combating post-inflammatory hyperpigmentation according to claim 3, characterized in that, In the steps S4 and S5, the filtration is microfiltration membrane filtration, the pore size of the microfiltration membrane is 0.1-1 μm, and the pore size of the ultrafiltration membrane in the step S7 is 0.01-0.05 μm.

7. Use of a composition according to any one of claims 1 to 2 to combat post-inflammatory hyperpigmentation of the skin, characterized in that, The composition is applied to cosmetics.

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