Composition for repairing cell light damage

Abstract

The invention belongs to the technical field of cosmetics, and particularly relates to a composition for repairing cell light damage and a preparation method and application thereof. The composition contains 0.01%-2% by mass of rice chaff extract and at least one antioxidant active matter. The composition containing the antioxidant active matter and the rice chaff extract has a definite synergistic effect in repairing cell light damage.

Description

Technical Field

[0001] The present invention belongs to the technical field of cosmetics, and in particular relates to a composition for repairing photodamage of cells, and a preparation method and application thereof. Background Art

[0002] Sunlight exposure can lead to many negative consequences, and the damage is related to its wavelength. Current research shows that harmful light mainly includes but is not limited to: ultraviolet rays and blue light; among them, the light with greater impact on the human body includes: UVA and UVB. UVB is mainly related to DNA damage and sunburn; UVA is mainly related to DNA oxidation and immunosuppression, and tanning; these two types of ultraviolet rays not only cause short-term damage to the skin, but also cause long-term damage. For example, short-term damage to the skin by UVA includes but is not limited to melanin deposition, darkening of the skin, and the production of spots; its long-term damage includes: damage to cell DNA, damage to collagen, skin aging, and even skin cancer; in contrast, UVB damage to the skin is concentrated on the cellular level, including: sunburn, tanning, erythema, edema, pain and even blisters (short-term) and damage to cell DNA (long-term); another component in sunlight that can cause skin damage is called: blue light, which is also called high-energy visible light (HIGHENERGY VISIBLE LIGHT), which is an important part of sunlight, can directly reach the basal layer of the skin, has strong penetration, and has a relatively long-lasting negative impact on the skin.

[0003] Interleukin 17 (IL-17) is one of more than 30 interleukins that have been discovered. It is secreted by CD4+T cells and can induce epithelial cells, endothelial cells, and fibroblasts to synthesize and secrete IL-6, IL-8, G-CSF, and PGE2, and promote the expression of ICAM-1.

[0004] There are more and more studies on damage caused by blue light. Generally speaking, the more common targets reported by blue light are photoaging target TRPV1 and pigmentation target OPN3. This patent discovered the IL-17 signaling pathway and its corresponding anti-inflammatory and anti-aging downstream targets for the first time through RNAseq sequencing, and obtained related compositions that can regulate the above signaling pathways. Summary of the invention

[0005] In order to achieve the above-mentioned purpose and overcome the deficiencies in the prior art, the present invention provides a composition for repairing photodamage of cells, and a preparation method and application thereof.

[0006] The present invention first discovered that a mixture obtained by mixing rice bran extract with one or both of the antioxidant composition ascorbic acid polypeptide or scutellaria root extract has the effect of resisting light damage and repairing, and applying the mixture to cosmetics can obtain cosmetics with the effect of resisting light damage and repairing. The present invention provides a cosmetic formula with a formulated ratio, and performs a corresponding light damage repair experiment on the cosmetic formula, verifying that the relevant composition of the present invention has a significant light damage repair effect.

[0007] In order to achieve the above-mentioned purpose and overcome the deficiencies in the prior art, the present invention provides a composition for repairing photodamage of cells, a preparation method and application.

[0008] In one aspect, the present invention provides a composition for repairing photodamage of cells, comprising 0.01% to 2% by weight of a rice bran extract and at least one antioxidant active substance, and further comprising water.

[0009] In some embodiments, the amount of the rice bran extract is preferably 0.025%-1% by mass, most preferably 0.025% by mass.

[0010] In some embodiments, the antioxidant active substance is one or both of ascorbic acid polypeptide (a solution material containing 1000 ppm EAC-L-Carnosine) or Scutellaria baicalensis root extract.

[0011] In some embodiments, the mass fraction of the ascorbic acid polypeptide is 0.001%-2%, preferably: 0.01%-1% by mass, and most preferably 0.05% by mass;

[0012] In some embodiments, the mass fraction of the scutellaria root extract is 0.001%-2%, and the dosage is preferably 0.005%-0.1% by mass, and most preferably 0.005% by mass.

[0013] In another aspect, the present invention also relates to the use of the composition for preparing cosmetics.

[0014] In some embodiments, the dosage form of the cosmetic is selected from one of cream, ointment, powder, lotion, and transparent liquid.

[0015] On the one hand, the present invention provides a composition for repairing photodamage of cells, characterized in that the composition comprises 0.01%-2% by mass of rice bran extract, and also comprises one or both of 0.01%-2% by mass of ascorbic acid polypeptide and 0.001%-2% by mass of Scutellaria baicalensis root extract.

[0016] In some embodiments, the amount of the rice bran extract is preferably: 0.025%-1%, for example: 0.025%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1% by mass.

[0017] In some embodiments, the composition includes 0.01%-1% by mass of rice bran extract, 0.005%-1% by mass of ascorbic acid polypeptide, and 0.001%-0.02% by mass of Scutellaria baicalensis root extract.

[0018] In some embodiments, the amount of the ascorbic acid polypeptide is preferably: 0.05%-1%, for example: 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1% by weight.

[0019] In some embodiments, the dosage of the Scutellaria baicalensis root extract is preferably: 0.005%-0.01%, for example: 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01% by weight.

[0020] In some embodiments, the dosage of the Scutellaria baicalensis root extract is preferably: 0.005%-1%, for example: 0.005%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1% by mass.

[0021] In some embodiments, the ratio of the rice bran extract: ascorbic acid polypeptide: scutellaria baicalensis root extract is 5:10:1-250:100:1.

[0022] In some preferred embodiments, the ratio of the rice bran extract: ascorbic acid polypeptide: scutellaria root extract is preferably 5:10:1 or 250:100:1.

[0023] In some embodiments, the ratio of rice bran extract: ascorbic acid polypeptide is 1:2-2.5:1.

[0024] In some preferred embodiments, the ratio of rice bran extract to ascorbic acid polypeptide is 1:1.

[0025] In some more preferred embodiments, the composition of the present invention consists of 0.01%-0.05% by weight of rice bran extract, 0.05%-1% by weight of ascorbic acid polypeptide, and 0.001%-0.01% by weight of Scutellaria baicalensis root extract and water.

[0026] In some most preferred embodiments, the composition of the present invention consists of 0.025% by mass of rice bran extract, 0.05% by mass of ascorbic acid polypeptide, 0.005% by mass of Scutellaria baicalensis root extract and water.

[0027] In some preferred embodiments, the composition for repairing photodamage of cells comprises 0.025% by mass of rice bran extract, 0.05% by mass of ascorbic acid polypeptide and 0.005% by mass of Scutellaria baicalensis root extract.

[0028] In some most preferred embodiments, the composition of the present invention consists of 0.025% by mass of rice bran extract, 0.05% by mass of ascorbic acid polypeptide, 0.005% by mass of Scutellaria baicalensis root extract and water.

[0029] In some preferred embodiments, the composition for repairing photodamage of cells comprises 0.25% by mass of rice bran extract, 0.1% by mass of ascorbic acid polypeptide and 0.001% by mass of Scutellaria baicalensis root extract.

[0030] In some preferred embodiments, the composition of the present invention consists of 0.25% by mass of rice bran extract, 0.1% by mass of ascorbic acid polypeptide, and 0.001% by mass of scutellaria root extract and water.

[0031] In some preferred embodiments, the composition of the present invention consists of 0.05% by mass of rice bran extract and 0.05% by mass of ascorbic acid polypeptide and or consists of 0.025% by mass of rice bran extract and 0.025% by mass of ascorbic acid polypeptide.

[0032] In another aspect, the present invention provides use of the composition in preparing cosmetics / skin care products / drugs for repairing photodamage.

[0033] In another aspect, the present invention provides use of the aforementioned composition for preparing cosmetics.

[0034] In another aspect, the present invention provides a cosmetic comprising the aforementioned composition.

[0035] Preferably, the dosage form of the cosmetic is selected from one of cream, ointment, powder, lotion, and transparent liquid. Preferably, the cosmetic is selected from any one of body lotion, face cream, eye cream, sunscreen, facial mask, facial mask liquid, essence, BB cream, CC cream, essence, liquid foundation, etc.

[0036] On the other hand, the present invention also provides a medicine containing the aforementioned composition;

[0037] In some embodiments, the medicament further comprises excipients and adjuvants.

[0038] In some embodiments, the excipient is selected from: one or more of hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyethylene glycol, polyacrylamide, polyacrylic acid, polyvinyl alcohol, dextran, lactic acid, citric acid, pyrrolidone carboxylic acid, glycerol, urea, sorbitol, amino acids, lanolin, and petrolatum.

[0039] In some embodiments, the adjuvant is selected from: one or more of a solubilizer, a thickener, a gelling agent, a softener, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a flavoring agent, a surfactant, water, an ionic or non-ionic emulsifier, a chelating agent, a preservative, a vitamin, a retardant, a wetting agent, an essential oil, a dye, and a pigment.

[0040] On the other hand, the present invention also relates to a method for preparing a rice bran extract, comprising:

[0041] The rice bran was dried and soaked in 50% propylene glycol for 3 days, and then the supernatant was filtered to obtain the rice bran extract.

[0042] On the other hand, the present invention also relates to the use of the composition with anti-photodamage and repair effects in the preparation of IL-17 inhibitors and / or IL-17 signaling pathway inhibitors.

[0043] On the other hand, the present invention also relates to the use of the composition for repairing photodamage of cells in the preparation of anti-aging and / or antioxidant agents.

[0044] On the other hand, the present invention also relates to use of the composition for repairing photodamage of cells in preparing a preparation for protecting fibroblast collagen from degradation caused by UVA.

[0045] In some embodiments, light damage repair includes protecting against cell damage caused by UVB. In some embodiments, light damage repair includes protecting against cell damage caused by blue light, and the main action pathway is the IL-17 pathway of keratinocytes.

[0046] In some embodiments, photodamage repair includes protection against UVA-induced fibroblast collagen degradation.

[0047] After years of research, the inventors of this application have obtained a composition that can repair cell photodamage (for example: blue light damage repair), and its main active ingredients involve: rice bran extract, scutellaria root extract and ascorbic acid polypeptide. On the one hand, experimental data have confirmed that rice bran extract and ascorbic acid polypeptide have significant synergistic effects in light damage repair (for example: resistance to cell damage repair caused by UVB). On the other hand, through comprehensive transcriptomics, genomics, and sequencing library construction and other molecular biological technical means, it is confirmed that the above-mentioned composition repairs the damage caused by blue light irradiation of HaCaT cells by affecting multiple pathways. Among them, the most significant change is the downregulation of the IL-17 signaling pathway. Therefore, it is confirmed that blue light damage to the skin can be repaired by regulating the IL-17 signaling pathway, and unexpected technical effects have been achieved.

[0048] In addition, the inventors of the present application have found through research that a composition containing ascorbic acid polypeptide and rice bran extract has a clear synergistic effect in repairing cell photodamage, and has achieved unexpected technical effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1 :The synergistic effect of ascorbic acid peptide and rice bran extract in repairing UVB damage. A = ascorbic acid peptide B = rice bran extract

[0050] Figure 2 : Transcriptomics sequencing workflow.

[0051] Figure 3 : Library construction process.

[0052] Figure 4 : Scatter plot and volcano plot of differentially expressed genes between the HaCaT group treated with the composition (treat group) and the group not treated with the composition (model group) after blue light irradiation.

[0053] Figure 5 : Heat map of differential gene clustering between the group treated with the composition (treat group) and the group not treated with the composition (model group) after HaCaT was irradiated with blue light.

[0054] Figure 6 : GO function analysis bar graph of HaCaT treated with the composition group (treat group) and the group not treated with the composition group (model group) after blue light irradiation.

[0055] Figure 7 : GO functional analysis column gene list of the group treated with the composition (treat group) and the group not treated with the composition (model group) after HaCaT was irradiated with blue light.

[0056] Figure 8: Expression content of SLC family genes in the composition-treated group (treat group) after HaCaT was irradiated with blue light.

[0057] Fig. 9 : Expression levels of MMP family genes in the combination-treated group (treat group) after HaCaT was irradiated with blue light.

[0058] Fig.10 : KEGG analysis gene list of differentially expressed genes in the combination-treated group (treat group) and the group not treated with the combination (model group) after HaCaT was irradiated with blue light.

[0059] Fig.11 : KEGG pathway enrichment of differentially expressed genes in the combination-treated group (treat group) and the group not treated with the combination (model group) after HaCaT was irradiated with blue light.

[0060] Fig.12 : Effect of the combination treatment group (treat group) on the IL-17 signaling pathway in HaCaT cells after blue light irradiation.

[0061] Fig.13 : Effect of the combination treatment group (treat group) on the expression of genes related to the IL-17 signaling pathway in HaCaT after blue light irradiation.

[0062] Fig.14 : Experimental effect diagram of the composition in repairing collagen degradation caused by UVA. DETAILED DESCRIPTION

[0063] definition

[0064] Although the numerical ranges and parameter approximations shown in the broad scope of the present invention, the numerical values ​​shown in the specific embodiments are recorded as accurately as possible. However, any numerical value is bound to contain a certain error, which is caused by the standard deviation in their respective measurements. In addition, all ranges disclosed herein should be understood to cover any and all sub-ranges contained therein. For example, the range of "1 to 10" recorded should be considered to include any and all sub-ranges between the minimum value 1 and the maximum value 10 (including the endpoints); that is, all sub-ranges starting with a minimum value of 1 or greater, such as 1 to 6.1, and sub-ranges ending with a maximum value of 10 or less, such as 5.5 to 10. In addition, any reference referred to as "incorporated herein" should be understood to be incorporated in its entirety.

[0065] Amino acids referred to in this application are represented herein by standard three-letter or single-letter abbreviations.

[0066] Example 1: Synergistic effect of ascorbic acid polypeptide and rice bran extract in repairing cell light damage

[0067] This experiment aims to evaluate the repair effects of the test products ascorbic acid peptide, rice bran extract, and ascorbic acid peptide + rice bran extract by analyzing. The zebrafish tail fins damaged by UVB were treated with 0.05% ascorbic acid peptide, 0.025% ascorbic acid peptide, 0.05% rice bran extract, 0.025% rice bran extract, 0.025% ascorbic acid peptide + 0.025% rice bran extract, 0.0125% ascorbic acid peptide + 0.0125% rice bran extract, water treatment control (Control) and untreated blank control (Blank), instrumental test and analysis of the zebrafish tail fin area, and comparison between the test product and the control group to verify the repair efficacy of the product. The specific steps are as follows:

[0068] 1. Collect zebrafish embryos that develop normally on the second day after fertilization, treat the embryos with different concentrations of the test substances for 24 hours, and expose them to UVB at a setting of 8100mJ / cm 2 (9mW / cm 2 , 15min). It includes a water treatment control group, an ascorbic acid polypeptide treatment group (0.05%, 0.025%), a rice bran extract treatment group (0.05%, 0.025%), and an ascorbic acid polypeptide + rice bran extract treatment group (0.025%, 0.0125%). In addition, an untreated blank control is set up without UVB exposure treatment. The preparation method of the above rice bran extract and ascorbic acid polypeptide is shown in Example 2.

[0069] 2. After UVB exposure, the test substance was incubated for 48 hours. Five zebrafish were randomly selected from each group on the fifth day after fertilization to observe their tail fins under a dissecting microscope. The images were analyzed to calculate the tail fin area. The experimental results are as follows:

[0070]

[0071] Image analysis and statistical analysis software were used to analyze the tail fin area of ​​each group. Figure 1). The results showed that the ascorbic acid polypeptide treatment group (0.05%, 0.025%), the rice bran extract treatment group (0.05%, 0.025%), and the ascorbic acid polypeptide + rice bran extract treatment group (0.025%, 0.0125%) all had significant repair effects on damage after UVB exposure. Among them, the tail fin repair effect of the 0.025% ascorbic acid polypeptide + 0.025% rice bran extract treatment group was significantly higher than that of the 0.05% ascorbic acid polypeptide and 0.05% rice bran extract treatment groups; the tail fin repair effect of the 0.0125% ascorbic acid polypeptide + 0.0125% rice bran extract treatment group was significantly higher than that of the 0.025% ascorbic acid polypeptide and 0.025% rice bran extract treatment groups.

[0072] The widely used Burgi formula method is used to calculate synergy: q = E〔A / 2+B / 2〕 / EA (or EB), where E〔A / 2+B / 2〕 refers to the effect of half the dose of A and B, and EA and EB are the effects of A and B alone. The q calculated by the above formula is greater than 1, which proves that the two have a synergistic effect.

[0073] In summary, the ascorbic acid polypeptide and rice bran extract of the present application have a clear synergistic effect in repairing ultraviolet damage and have achieved unexpected technical effects.

[0074] Example 2: Preparation of a composition for repairing photodamage of cells

[0075] 1. Dry the rice bran and soak it in 50% propylene glycol (mass ratio: rice bran: propylene glycol = 1:7). After soaking for 3 days, filter and take the supernatant to obtain the rice bran extract;

[0076] 2. Ascorbic acid polypeptide: The synthesis route of ascorbic acid polypeptide (containing 1000 ppm EAC-L-Carnosine, 3-O-ethyl ascorbic acid-carnosine) is as follows (the entire content of WO2022 / 228475A1 patent is cited in this application): comprising the following steps:

[0077]

[0078] ①, Boc-L-Carnosine (13 g, 39.9 mmol) was dissolved in DMF (400 mL);

[0079] ②, add DCC (7.98 g, 38.8 mmol) and HOSu (4.45 g, 38.8 mmol);

[0080] ③. Stir the solution at room temperature for about 15 hours, and filter to remove the solid by-product (N,N-dicyclohexylurea).

[0081] ④. Add 3-O-ethyl ascorbic acid ether (8.2 g, 40.2 mmol) and DIPEA (16 mL, 92 mmol) to the filtrate;

[0082] ⑤. The mixture was stirred at room temperature for 3 h, then concentrated and dried in vacuo;

[0083] ⑥. The solid was recrystallized from methanol and isopropyl ether to obtain Boc-EAC-L-Carnosine (16.4 g, 32 mmol) as a light yellow solid;

[0084] ⑦, Boc-EAC-L-Carnosine was dissolved in 1,4-dioxane (100 mL), and 4M hydrochloric acid and 1,4-dioxane (99 mL, 396 mmol) were added;

[0085] ⑧. The mixture was stirred at room temperature for 4 h, then concentrated, and the solid was triturated with acetone and isopropanol to purify EAC-L-Carnosine (3-O-ethyl ascorbic acid-carnosine) (5.2 g, 98%);

[0086] ⑨. After freeze-drying, EAC-L-Carnosine is a white powder (yield 4.9%).

[0087] Note:

[0088] DCC = dicyclohexylcarbodiimide; HOSu = N-hydroxysuccinimide

[0089] DIPEA = diisopropylethylamine

[0090] 3. Prepare a mixed solution containing 0.005% scutellaria root extract (Shandong Mingren Furuida Pharmaceutical Co., Ltd., scutellaria root purity> 85%) and 0.05% ascorbic acid polypeptide. Then use the solution to prepare a rice bran extract solution so that its final concentration is 0.025% to obtain an anti-blue light composition. Therefore, the anti-blue light composition finally obtained contains the following ingredients: 0.05% ascorbic acid polypeptide, 0.005% scutellaria root extract, and 0.025% rice bran extract.

[0091] Example 3: Transcriptomic sequencing analysis of blue light-induced HaCaT cell injury model

[0092] The basic analysis steps of transcriptomics are as follows:

[0093] 1. RNA extraction: discard the culture supernatant and collect the cell pellet, then extract RNA according to the instructions of the Trizol kit and determine the RNA concentration.

[0094] 2. Construction of sequencing library. The library construction process is as follows: Figure 2 , Figure 3 shown.

[0095] (1) Construction of mRNA library

[0096] 1) Use Qubit 2.0 RNA detection kit to accurately quantify total RNA.

[0097] 2) Purification and fragmentation of mRNA

[0098] After enrichment of mRNA, the mRNA is broken down into short fragments.

[0099] 3) Synthesis of double-stranded cDNA

[0100] The interrupted mRNA is used as a template to synthesize the first-strand cDNA, and then a second-strand synthesis reaction system is prepared to synthesize the second-strand cDNA.

[0101] 4) Purification of double-stranded cDNA

[0102] Hieff NGSTM DNA Selection Beads DNA purification of second strand cDNA.

[0103] 5) End repair / dA tail addition

[0104] The fragmented dscDNA, Endprep Buffer and Endprep Enzyme were placed in a PCR instrument for end repair / dA tailing reaction.

[0105] 6) Connector connection

[0106] dA-tailed DNA, Ligation Enhancer, Quick T4 DNALigase and DNA Adapter were placed in a PCR instrument for adapter ligation reaction.

[0107] 7) Purification of ligation product

[0108] 8) Library amplification

[0109] 2×Super Canace TM High-Fidelity Mix, Primer mix and Adapter Ligated DNA were placed in a PCR instrument for PCR amplification, and then the library amplification products were purified using Hieff NGSTM DNA Selection Beads.

[0110] (2) Library quality control

[0111] Detection was performed by gel electrophoresis.

[0112] (3) Quantitative mixing

[0113] Accurately quantify DNA and mix equal amounts in a 1:1 ratio.

[0114] Raw data processing and analysis:

[0115] The fastq sequence files were analyzed for quality using fastQC software, and then the adapters in the sequence files were removed using Trimmomatic software, and the bases were appropriately modified, and then the bases were trimmed and low-quality sequences were filtered. The fasta sequences were aligned to the reference genome using hisat2 software. The reads were then accurately counted using featuresCounts software, and finally the reads of all samples were merged into one file, and the RNA-Seq_Practice_countstable file was exported. The expression of each gene in each sample was quantified according to the calculation formula of FPKM and TPM. The gene ID was converted to Gene description using the website (https: / / www.ncbi.nlm.nih.gov / gene) to understand its function and related information. The generated RNA-Seq_Practice_countstable was saved locally, and then the FPKM and TPM values ​​were calculated, and related calculations were performed in R language. DESeq2 software was used to calculate differentially expressed genes based on the original reads number of the samples, and the run-featurecounts.R script was used to quantify the number of reads of each sample. Then, the abundance_estimates_to_matrix.pl script was used to merge all the quantitative results, and finally DESeq2 was used to analyze the differentially expressed genes.

[0116] GO enrichment analysis was performed using an online website (https: / / www.metascape.org), and the input file was the ID information table of differentially expressed genes.

[0117] KEGG enrichment analysis was performed using three R packages: clusterProfiler, KEGG.db, and ggplot2 in R language (4.2.2).

[0118] Transcriptomics results: A total of 6 cDNA libraries were constructed on the Illumina Casava 1.8 platform, and the raw data were filtered to obtain Clean data. The Q20 Bases Ratio (%) of the Clean data was >97%, and the Q30 Bases Ratio (%) was >91%, indicating that the sequence results obtained by sequencing were of high quality. 10,000 sequences were randomly selected from the Clean data and blastn compared with the NCBINT database. The results showed that Homo sapiens accounted for more than 96.5%, and there was no obvious contamination in the samples.

[0119] 3. Determination of Differentially Expressed Genes

[0120] The screening conditions for differentially expressed genes were: p-Value < 0.05, fold change |FoldChange| ≥ 2, and the expression value was ≥ 5 in at least one of the two comparison groups.

[0121] After HaCaT cells were irradiated with blue light, a total of 646 genes with significant differences were identified between the group treated with the anti-blue light composition (containing 0.025% rice bran extract, 0.05% ascorbic acid polypeptide, 0.005% scutellaria root extract, and the rest was water) and the group not treated with the composition (P<0.05), of which 134 were up-regulated genes and 138 were down-regulated genes. The scatter plot and volcano plot of the differential gene distribution are shown in Figure 2. Figure 4 The heat map of genes with a difference of more than 5 is shown in Figure 5 shown.

[0122] 4. GO analysis of up-regulated and down-regulated differentially expressed genes

[0123] GO (Gene Ontology) has a total of three ontologies involving 67 subcategories, which can be used to clarify the main biological functions of differentially expressed genes.

[0124] After HaCaT cells were irradiated with blue light, GO analysis was performed in the group treated with the anti-blue light composition (containing 0.025% rice bran extract, 0.05% ascorbic acid peptide, and 0.005% scutellaria root extract) and the group not treated with the composition. Figure 6 and Figure 7 As shown in the figure, the up-regulated genes and down-regulated genes were involved in 15 biological functions, thus affecting HaCaT after blue light irradiation. Among them, the anti-blue light composition can up-regulate genes related to the biological function of cell plasma membrane transport ( Figure 8 ) and down-regulate genes related to matrix metalloproteinase synthesis and collagen degradation ( Fig. 9), thereby accelerating the transport of active factors such as metal ions, organic anions, and amino acids in the human body, further inhibiting NF-κB, promoting GSH synthesis, protecting cells from oxidative stress damage caused by blue light irradiation, maintaining cell redox balance, and preventing cell death caused by lipid peroxidation. It can also reduce the degradation of collagen and elastin in skin cells after blue light irradiation, thereby delaying aging.

[0125] 5. KEGG analysis of up-regulated and down-regulated differentially expressed genes

[0126] KEGG (Kyoto Encyclopedia of Genes and Genomes) is a database that integrates genome, chemical substances and system function information. Different genes in organisms coordinate with each other to form pathways to exert their biological functions. Enriching differentially expressed genes in pathways is conducive to a systematic and comprehensive understanding of the physiological functions exerted by differentially expressed genes.

[0127] After HaCaT cells were irradiated with blue light, KEGG analysis of up- and down-regulated genes in the group treated with the anti-blue light composition (containing 0.05% ascorbic acid peptide, 0.005% scutellaria root extract, and 0.025% rice bran extract) and the group not treated with the composition was as follows: Fig.10 and Fig.11 As shown. After treatment with the anti-blue light composition, the upregulated differentially expressed genes were mainly enriched in pathways related to rheumatoid arthritis, cytokine-cytokine receptor interaction, hypercortisolism, MAPK, etc., while the downregulated genes were mainly enriched in signaling pathways such as IL-17 signaling pathway, rheumatoid arthritis, TNF signaling, and cofactor biosynthesis. This shows that the anti-blue light composition repairs the damage caused by blue light irradiation to HaCaT cells by affecting multiple pathways. Among them, the most significant change was the downregulation of the IL-17 signaling pathway.

[0128] Based on the above data, the effects of the anti-blue light composition on the IL-17 signaling pathway are as follows:

[0129] The IL-17 family is a subset of cytokines composed of IL-17A-F. IL-17 mediates the production of inflammatory mediators and mobilizes immunosuppressive and angiogenic myeloid cells. In addition, studies have shown that IL-17 secreted by skin cells promotes inflammation and accelerates skin aging. IL-17 can also stimulate the production of a variety of chemokines, including CXCL1, CCL2, CXCL5, etc. These chemokines can recruit monocytes and neutrophils to the synovial membrane of the joints, thereby aggravating inflammation. IL-17 can also cause extracellular matrix degradation by inducing matrix metalloproteinases (MMP) 1, 2, 3, 9, and 13, while inhibiting the synthesis of extracellular matrix, ultimately making the skin barrier repair ability worse while reducing the skin's ability to repair damage, and at the same time, epidermal cells are enriched with various inflammatory factors. Therefore, blocking the IL-17-mediated signaling pathway helps to delay skin inflammation associated with skin aging and improve the characteristics of aging skin. The regulation of the IL-17 signaling pathway by the anti-blue light composition is as follows: Fig.12 After treatment with the anti-blue light composition, the gene expression of the trending factors CXCL1, CCL2, and CXCL5 was significantly reduced, thereby inhibiting the aggregation of immune cells and reducing the inflammatory response (see Fig.13 ). In addition, after treatment, the expression of MMP-1, MMP-3 and MMP-13 genes decreased. Therefore, the anti-blue light composition can reduce the inflammatory response and aging of HaCaT cells caused by blue light irradiation by inhibiting the secretion of downstream trend factors and matrix metalloproteins in the IL-17 signaling pathway.

[0130] Example 4: Effect of the composition in repairing collagen degradation caused by UVA

[0131] The steps for determining the content of type I collagen by ultraviolet light treatment are as follows:

[0132] Move the fibroblasts (HFF cells) to a 6-well plate. After 24 hours of cell inoculation, the fibroblast fusion rate reaches 45% to 65%. Remove the original culture medium and replace it with PBS containing the sample to be tested. Place the cells in a carbon dioxide incubator at 37°C for 24 hours ± 2 hours, and the UVA radiation dose is 5J / cm 2 After the incubation, 200 μL of cell culture supernatant was taken from each well, placed in a 1.5 mL sterile centrifuge tube, and stored in a -80°C ultra-low temperature refrigerator. First, the standard was used to detect and draw a standard curve, and then the amount of collagen contained in the fibroblast supernatant diluted in an appropriate ratio was detected. ELISA detection was performed using the Human Type I Collagenase Immunoassay Kit (Cusabio, China).

[0133] in,

[0134] Blank control: untreated normal HFF cells

[0135] Negative control: 15J / cm 2 UVA-treated HFF cells

[0136] Positive control: HFF cells treated with UVA + TGF-β1 (100 ng / mL)

[0137] Composition to be tested: HFF cells treated with UVA + composition (containing 0.25% by mass of rice bran extract, 0.1% by mass of ascorbic acid polypeptide and 0.001% by mass of scutellaria root extract, and the rest being water).

[0138] The experimental results show that (see Fig.14 ): The composition claimed in the present application is significantly better than the control and 0.5% rice bran extract in resisting collagen degradation caused by UVA, and has achieved unexpected technical effects.

Claims

1. A composition for repairing photodamage to cells, characterized in that: The composition contains 0.01% to 2% by weight of rice bran extract and at least one antioxidant active substance.

2. The composition according to claim 1, characterized in that The amount of the rice bran extract is 0.025%-1% by weight.

3. The composition according to claim 1, characterized in that The antioxidant active substance is selected from one or both of ascorbic acid polypeptide and scutellaria baicalensis root extract; the ascorbic acid polypeptide is a solution raw material containing 1000 ppm EAC-L-Carnosine.

4. The composition according to claim 3, characterized in that The mass fraction of the ascorbic acid polypeptide is 0.001%-2%, wherein the synthesis path of EAC-L-Carnosine is as follows:

5. The composition according to claim 3, characterized in that The mass fraction of the scutellaria root extract is 0.001%-2%.

6. The composition according to claim 1, characterized in that The composition comprises 0.05% by mass of rice bran extract and 0.05% by mass of ascorbic acid polypeptide; or, the composition comprises 0.025% by mass of rice bran extract and 0.025% by mass of ascorbic acid polypeptide.

7. Use of the composition according to any one of claims 1 to 6 for preparing cosmetics.

8. The use according to claim 7, characterized in that The dosage form of the cosmetic is selected from one of cream, ointment, powder, emulsion and transparent liquid.

9. A medicine comprising the composition according to any one of claims 1 to 6.

10. Use of the composition in preparing a drug for repairing photodamage and / or in preparing a cosmetic or skin care product for repairing photodamage.

11. The use of the composition according to claim 10, characterized in that: The photodamage repair includes the repair of cell damage caused by UVB.

12. The use of the composition according to claim 10, wherein the photodamage repair comprises protecting against cell damage caused by blue light, and the main action pathway is the IL-17 pathway of keratinocytes.

13. The use of the composition according to claim 10, wherein the photodamage repair comprises protecting against fibroblast collagen degradation caused by UVA.

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