Homologous bioactive peptide of yeast fermentation product and application of homologous bioactive peptide

Abstract

The invention relates to the field of biological medicines and daily chemicals, and discloses a bioactive peptide, the amino acid sequence of which is ETRIL, or hydroxylation, carboxylation, carbonylation, methylation, acetylation, phosphorylation, esterification or glycosylation modification is carried out on the amino acid side chain group, the amino terminal or carboxyl terminal of the bioactive peptide, and the obtained derivative has the same biological activity. The bioactive peptide has good anti-aging, anti-oxidation, sunscreen and other skin care effects, is homologous with a yeast fermentation product, can be used in the fields of medicines, health care products, food, daily chemical products and the like, and has a good application prospect.

Description

Technical Field

[0001] This application relates to the fields of biomedicine and daily chemicals. Specifically, it relates to the application of a bioactive peptide with anti-aging activity, as well as skin care products or drugs with this bioactive peptide as the main ingredient. Background Art

[0002] Bioactive peptides are a class of polypeptides beneficial to the health of the body or with certain specific biological functions, and are specific protein fragments composed of short amino acid sequences; generally composed of 2 or more and less than 20 amino acids polymerized by peptide bonds, with a molecular weight below 6 kDa; due to their special biological activity, they become the basic components of protein structure and function. They can be obtained from protein hydrolysates by means of enzymatic hydrolysis, microbial fermentation, acid and alkali treatment, etc. Bioactive peptides have a wide range of biological activities, including antioxidant, ACE inhibition, anti-tumor, immunomodulatory, anti-aging and anti-inflammatory, etc. Due to the characteristics of high specificity, low toxicity, high structural diversity and low molecular weight of bioactive peptides, they are widely used in industries such as medicine, food, and cosmetics.

[0003] Skin aging is an external manifestation of human aging. Research shows that the main cause of skin aging is that oxygen free radicals generated by factors such as ultraviolet rays affect the normal growth cycle of the skin, promote the hydrolysis of extracellular matrix such as collagen by proteases, and cause DNA damage resulting in a decrease in the synthesis of matrix proteins, ultimately leading to skin relaxation and reduced elasticity. Since the 1990s, researchers have tried to prevent and modify the skin physiological structure or exogenous factors causing skin aging by adding biological macromolecules such as DNA and proteins. However, it is difficult for these macromolecular active ingredients to penetrate the epidermis and be absorbed. Current research shows that small molecule bioactive peptides have the characteristics of good safety and high biological activity, and are more likely to penetrate the skin barrier. Polypeptides with anti-aging activity can control or mediate the skin aging process, thereby protecting the damaged skin and delaying skin aging.

[0004] HiMurchaSin ® (HiMurchaSin ® ) is a fermentation product of a Saccharomyces cerevisiae strain derived from the Himalayas (CN114081862A, the preservation number in the China General Microbiological Culture Collection Center is CGMCC No. 19732), containing various ingredients beneficial to the skin, such as proteins, peptides, amino acids, minerals and vitamins, etc., with excellent repair ability and cell renewal ability, and can promote the proliferation of skin cells and the regeneration of collagen, thus achieving the effects of anti-aging and repair. By studying the protein and polypeptide information identified in the HiMurchaSin sample, if bioactive peptides with skin care effects, especially short peptides with a smaller molecular weight, can be found, it will play an important role in promoting skin absorption, improving the anti-aging activity or other skin care effects of the product. Summary of the Invention

[0005] Based on the deficiencies of the prior art, the present invention provides a bioactive peptide with anti-aging effects and its applications.

[0006] In a first aspect, the present invention relates to a bioactive peptide EL-5, whose amino acid sequence is ETRIL, as shown in SEQ ID No. 1.

[0007] The bioactive peptide EL-5 is homologous to Hymecromone and is a short peptide obtained from the fermentation product of the Saccharomyces cerevisiae strain (CGMCC No. 19732) through experiments and bioinformatics analysis. Its molecular weight is about 700 Da, and it has good skin care effects such as anti-aging, antioxidant, and sunscreen.

[0008] In some embodiments, modifications such as hydroxylation, carboxylation, carbonylation, methylation, acetylation, phosphorylation, esterification, or glycosylation can be carried out on the amino acid side chain groups, amino terminus, or carboxyl terminus of the bioactive peptide to obtain derivatives of the bioactive peptide without affecting its biological activity.

[0009] The bioactive peptide EL-5 can promote the production of collagen, especially significantly increasing the content of type I collagen after damage caused by ultraviolet irradiation.

[0010] The bioactive peptide EL-5 can increase the expression and content of ki67 and increase the ATP content, and can promote cell proliferation and skin repair. Especially after induction by UVA or hydrogen peroxide, it can increase the expression of ki67 and promote cell proliferation. Therefore, it has anti-aging, sunscreen, and anti-photoaging effects.

[0011] The bioactive peptide EL-5 also has antioxidant effects, inhibiting hydroxyl radicals and oxygen radicals, especially reducing the content of ROS after induction by hydrogen peroxide.

[0012] The bioactive peptide EL-5 can also reduce the inflammatory response, specifically the inflammatory response induced by ultraviolet light, especially inhibiting the release of the IL-8 inflammatory factor induced by UVB.

[0013] In some embodiments, the N-terminus of the bioactive peptide is acetylated and the C-terminus is amidated.

[0014] In a second aspect, the present invention relates to a pharmaceutical composition comprising the above-mentioned bioactive peptide EL-5. Further, it also contains a pharmaceutically acceptable carrier and / or the fermentation product of the Saccharomyces cerevisiae with the preservation number of CGMCC No. 19732.

[0015] In some embodiments, the fermentation product is selected from any one or any combination of fermentation broth, fermentation filtrate, concentrated fermentation broth or fermentation filtrate, and dry powder of fermentation broth or fermentation filtrate.

[0016] In some embodiments, the pharmaceutically acceptable carrier is selected from any one or any combination of isotonic saline, ethanol, phosphate buffered saline, sorbitol, mannitol, starch, acacia gum, gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, and water.

[0017] In some other embodiments, the composition can be administered orally, buccally, topically, by inhalation, rectally, nasally, ophthalmically, or parenterally.

[0018] In some other embodiments, the dosage form of the composition includes any one or any combination of oral dosage form, parenteral dosage form, buccal dosage form, sublingual dosage form, nasal dosage form, inhaler, nebulizer, topical dosage form, transdermal dosage form, and suppository.

[0019] In a third aspect, the present invention relates to a daily chemical composition comprising the above-mentioned bioactive peptide EL-5.

[0020] In some embodiments, the daily chemical composition further contains a fermentation product of Saccharomyces cerevisiae with the preservation number of CGMCC No. 19732. The fermentation product is selected from any one or any combination of fermentation broth, fermentation filtrate, concentrated fermentation broth or fermentation filtrate, and dry powder of fermentation broth or fermentation filtrate.

[0021] In a fourth aspect, the present invention relates to the use of the above-mentioned bioactive peptide, pharmaceutical composition or daily chemical composition for preparing a drug, health product or food with anti-aging efficacy.

[0022] In some embodiments, the bioactive peptide, pharmaceutical composition or daily chemical composition can reduce collagen degradation and smooth fine lines; accelerate skin cell growth, inhibit oxygen free radicals and hydroxyl free radicals; and as a strong antioxidant, can inhibit photoaging; can also repair damaged skin, reconstruct the epidermal structure, and tighten the skin; reduce inflammatory responses and provide long-term soothing.

[0023] In some other embodiments, the bioactive peptide, pharmaceutical composition or daily chemical composition can increase the content of type I collagen, increase the expression of ki67, and increase the content of ATP; inhibit ROS generation, and antagonize IL-8 inflammatory factors, especially IL-8 inflammatory factors induced by UVB.

[0024] The bioactive peptide EL-5 and the fermentation product (Ximoyin) of Saccharomyces cerevisiae with the preservation number of CGMCC No. 19732 can produce synergistic effects.

[0025] EL-5 and the composition of the fermentation product of Hymecromone have a synergistic effect in terms of the ability to increase the content of Col I (type I collagen), especially significantly increasing the content of type I collagen after ultraviolet irradiation.

[0026] In a fifth aspect, the present invention relates to the use of the bioactive peptide or the pharmaceutical composition or the daily chemical composition for preparing skin care products or cosmetics. In particular, skin care products with anti-aging and sunscreen effects.

[0027] In some embodiments, the cosmetics or skin care products have anti-aging effects.

[0028] In some embodiments, the cosmetics or skin care products have the effects of antioxidant, sunscreen, and anti-photoaging.

[0029] In some embodiments, the cosmetics or skin care products have antioxidant effects and can inhibit hydroxyl radicals and oxygen radicals, especially ROS induced by hydrogen peroxide. The cosmetics or skin care products can increase the expression of ki67 and the content of ATP, promote the proliferation of skin cells; can also increase the content of type I collagen, promote the production and secretion of type I collagen, especially promoting the production of type I collagen after damage caused by UV irradiation. And, it has the effect of reducing inflammatory reactions and can inhibit the release of IL-8 inflammatory factors induced by UVB.

[0030] In a sixth aspect, the present invention relates to a method for preparing the bioactive peptide, which is prepared by genetic engineering, enzyme engineering or chemical synthesis methods.

[0031] The beneficial effects of the present invention are as follows: a bioactive peptide that promotes collagen secretion and cell proliferation, has better anti-inflammatory and antioxidant effects, and has good anti-aging, antioxidant, anti-photoaging and other effects is provided. This active peptide is derived from the identification of the protein spectrum of Hymecromone samples and is homologous to the yeast fermentation product; and compared with the prior art, this bioactive peptide has a shorter length, a smaller molecular weight, reduces the production cost on the basis of improved activity, and is more conducive to absorption. It can be used in the fields of pharmaceuticals, health products, foods, daily chemical products, etc., and has good application prospects. Description of the Drawings

[0032] Figure 1 It is the result of the biological activity test of EL-5 and EL-6 promoting the secretion of type I collagen by HFF-1 cells after ultraviolet irradiation.

[0033] Figure 2 It is the result of the biological activity test of EL-5 and Hymecromone promoting the secretion of type I collagen by HFF-1 cells after ultraviolet irradiation.

[0034] Figure 3 The test experimental results of EL-5 promoting the expression of ki67 in HFF-1 cells.

[0035] Figure 4 The test experimental results of EL-5 promoting the expression of ki67 in HaCaT cells.

[0036] Figure 5 The experimental results of EL-5 inhibiting ROS in FB cells.

[0037] Figure 6 The test experimental results of EL-5 increasing the ATP content of fibroblasts.

[0038] Figure 7 The test experimental results of EL-5 antagonizing IL-8 induced by UVB. Detailed implementation manners

[0039] Unless otherwise defined, all scientific and technical terms used herein have the same meaning as commonly understood by those of ordinary skill in the art. Regarding the definitions and terms in this field, professionals can specifically refer to Current Protocols in Molecular Biology (Ausubel). The abbreviations of amino acid residues are the standard 3-letter and / or 1-letter codes used in the art to refer to one of the 20 common L-amino acids.

[0040] Although the numerical ranges and parameter approximations shown in the broad scope of this application, the values shown in the specific embodiments are recorded as accurately as possible. However, any value inherently necessarily contains certain errors, which are caused by the standard deviations existing 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" 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 from the minimum value 1 or greater, such as 1 to 6.1, and sub-ranges ending with the maximum value 10 or less, such as 5.5 to 10. In addition, any reference cited as "incorporated herein" should be understood to be incorporated in its entirety.

[0041] Example 1 Obtaining of EL-5 peptide CN114081862A discloses a Saccharomyces cerevisiae from highland barley wine koji with the preservation number of CGMCC No. 19732, and its fermentation product (Ximoyin) has the functions of scavenging free radicals, promoting cell proliferation, inhibiting tyrosinase, protecting skin cells from damage caused by oxidation or ultraviolet rays, and regulating skin flora and microecology. Therefore, it can achieve the effects of antioxidant, anti-aging, sunscreen, improving skin cell vitality and protecting skin barrier, etc.

[0042] After component analysis, it contains various types of active ingredients such as organic acids, phenolic vitamins, free amino acids, peptides, and protein components.

[0043] Furthermore, mass spectrometry analysis was performed on the protein components from the fermentation products, and multiple candidate protein polypeptide molecules were obtained. The amino acid sequence of one polypeptide is ETRIL (abbreviated as EL-5), and its molecular weight is approximately 700 Da.

[0044] EL-5 can be prepared by genetic engineering, enzyme engineering, or chemical synthesis methods. The following are the tests for various effects of EL-5 respectively.

[0045] Example 2 Col I test experiment EL-5 and its analog EL-6 (amino acid sequence: ETRILL, SEQ ID No.2) were taken to detect their effects on promoting the production of type I collagen (Col I) in human fibroblasts (HFF-1) after ultraviolet irradiation, with TGF-β1 as the positive control.

[0046] (1) Plating: After digestion and resuspension of HFF-1 cells, the cell count was calculated using a counstar cell counter. Finally, the DMEM complete medium containing cells (WISENT, 319-010-CL) was added to a 24-well plate, and the final cell density was 8×10 4 . After plating, it was placed in a cell culture incubator and cultured for 24 h; (2) Induction and sample addition: Drugs were administered when the cell culture reached a confluence of about 40% - 60%. 1 mL of DMEM complete medium was added to each well in the blank control group and the model group; 1 mL of DMEM complete medium containing 100 ng / mL of TGF-β1 was added to each well in the positive control group; 1 mL of DMEM complete medium containing the corresponding concentrations (10 ppm, 25 ppm) of the test substance was added to each well in the sample group.

[0047] (3) Ultraviolet induction: 24 h after drug administration, the model group, the positive control group, and the sample groups (EL-6 / EL-5: 10 ppm and 25 ppm) received UVA radiation with a total dose of 8 mJ / cm 2 for 10 min 25 s. At the same time, the blank control group was placed in the same environment (UVA radiation dose of 0 J / cm 2 ).

[0048] (4) Sample collection: After incubation for 24 h, the cell culture supernatant was collected into an EP tube and stored frozen at -80 °C. And 0.5 mL of lysis buffer was added to each well and pipetted to lyse. The cell lysate was collected into an EP tube, and the samples collected for detection were stored frozen in a -80 °C refrigerator. Detection was performed according to the type I collagen content detection kit.

[0049] As Figure 1 , the experimental results show that: compared with the blank control group, the content of Col I in the model group after inducing cells decreased significantly; compared with the model group, the promotion rate of the positive control group (TGF-β1 100 ng / mL) was 354.88%; compared with the model group: 10 ppm EL-5 could significantly improve the decrease in ColI content caused by induction, and the promotion rate was 268.97%; the promotion rate of 25 ppm EL-5 was 338.18%. In addition, compared with EL-6, EL-5 with a shortened length under the same dosage significantly enhanced the ability to increase the ColI content, achieving an unexpected technical effect.

[0050] In step (2), the DMEM complete medium containing 0.1 ppm EL-5, 1 ppm EL-5, 5% (wt) Hymain (fermentation broth), 0.1 ppm EL-5 + 5% (wt) Hymain (fermentation broth, prepared according to the method of CN114081862A) was added to the sample groups respectively. After incubation, ultraviolet induction and sample collection were carried out according to the methods of steps (3) and (4) to detect their relative fluorescence intensity.

[0051] As Figure 2 , the experimental results show that: compared with the blank control group, the content of Col I in the model group after inducing cells decreased significantly; compared with the model group, the promotion rate of the positive control group (TGFβ1 100 ng / mL) was 329%. Compared with the model group: 0.1 ppm EL-5 could significantly increase the content of Col I to 125%, and 1 ppm EL-5 could significantly increase the content of Col I to 300%, exceeding the effect of 5% Hymain yeast fermentation product. At the same time, the combination of EL-5 and Hymain yeast fermentation product has a synergistic effect on the ability to increase the content of Col I.

[0052] The biological experimental results show that compared with the analogue EL-6 (sequence: ETRILL), EL-5 has a stronger ability to promote the secretion of type I collagen by HFF-1 cells after UVA damage. It has been confirmed that the functional peptide segment EL-5 is 100% matched with some fragments of the proteins identified in the Hymain sample, and then the anti-aging activity and anti-photoaging activity of the functional polypeptide EL-5 in Hymain are deeply explored.

[0053] Example 3 HFF cell ki67 test experiment Ki67 is a protein related to cell proliferation. By detecting the content of ki67 in HFF-1 (human skin fibroblasts), it was found that EL-5 could increase the content of ki67, especially after UVA induction.

[0054] Experimental materials: HFF-1 cells, ki67 content induced by UVA (UVA induction dose: 40 mJ / cm 2 ) Experimental method: The cell seeding culture and drug administration methods were similar to those in Example 2; 24 hours after drug administration, the model group, positive control group, and sample groups (EL-6 / EL-5: 10 ppm and 25 ppm) were irradiated with UVA with a total dose of 40 mJ / cm 2 for 30 minutes; meanwhile, the blank control group was placed in the same environment (UVA radiation dose: 0 J / cm 2 ).

[0055] The expression of ki67 was detected by immunofluorescence method. Specifically: For immunofluorescence staining of cells, cells were seeded in a confocal bottom glass dish. After the cells were co-cultured for 24 hours or 48 hours to form a vascular network structure, the following cell fixation and staining steps were carried out: 1) Discard the culture medium and wash the cells 3 times with PBS; 2) Add 4% paraformaldehyde and fix at room temperature for 15 minutes; 3) Wash again 3 times with PBS, 5 - 10 minutes each time; 4) Treat with 0.5% TritonX-100 for 3 - 5 minutes; 5) Block with PBS containing 5% FBS for 1 hour; 6) Dilute the primary antibody with PBS containing 5% FBS to 5 - 10 μg / ml and incubate overnight at 4°C; 7) Wash 3 times with PBS, 5 - 10 minutes each time; 8) Dilute the fluorescently labeled secondary antibody with PBS containing 5% FBS at a ratio of 1:100 and incubate in the dark at room temperature for 2 hours; 9) Wash once with PBS, add Hoechst 33342 or DAPI to PBS and stain in the dark for 15 minutes; Subsequently, wash 2 more times with PBS and perform fluorescence imaging observation or glycerol mounting.

[0056] Test groups: EL-5 (0.1 ppm, 1 ppm, 10 ppm) The results are as Figure 3, The experimental results showed that: compared with the blank control group (Control), the content of ki67 in HFF-1 cells induced by UVA decreased significantly, indicating that the model group (Model) was successfully established; compared with the model group, the ki67 content was increased by the EL-5 samples at three concentrations (154.22%, 314.14% and 453.16% respectively), showing a concentration-dependent manner. In conclusion, the EL-5 sample has a strong ability to enhance the expression of ki67 induced by UVA, can promote skin cell proliferation and repair the skin, especially after UVA irradiation, and thus has an anti-photoaging effect.

[0057] Example 4 Ki67 test experiment on HaCaT cells Experimental materials: Human immortalized keratinocytes (HaCaT cells, ki67 test after H2O2 induction (induction intensity: 200 μM, 6 h), H2O2 Test groups: EL-5 (0.1 ppm, 1 ppm, 10 ppm) Experimental method: The cell plating and culture method was the same as in Example 3; for induction and sample addition: when the cell plating rate reached 40% - 60%, grouping and drug administration were carried out. Each group had 3 replicate wells, and the volume of liquid added to each well was 2 mL. The blank control (Control) group added culture medium to each well, the model group (Model) added culture medium containing 200 μM H2O2 to each well, and the sample group added culture medium containing 0.1, 1, 10 ppm of EL-5 sample and H2O2 to each well. After 6 h, it was placed in a CO2 incubator (37 °C, 5% CO2) and continued to be cultured for 24 h.

[0058] The detection method was the same as in Example 3.

[0059] The experimental results are as Figure 4 shown. Compared with the blank control group (Control), the content of ki67 in cells induced by H2O2 decreased significantly, indicating that the model group (Model) was successfully established; compared with the model group, the ki67 content was increased by the EL-5 samples at three concentrations (98.19%, 153.82% and 251.29% respectively), showing a concentration-dependent manner. Thus, EL-5 has the ability to enhance the expression of ki67 induced by H2O2, and is therefore a strong antioxidant with an anti-aging effect.

[0060] Example 5 ROS test experiment on fibroblasts Experimental materials: Fibroblasts FB; positive control VE, sample group: 0, 1, 5 ppm EL-5.

[0061] Experimental method: (1)Cell seeding: After resuscitating fibroblast FB and when the seeding rate reaches about 60%, seed the cells into a 6-well plate and incubate overnight in a CO2 incubator (37 °C, 5% CO2).

[0062] (2)Induction and sample addition: When the seeding rate of the cells in the 6-well plate reaches 40% - 60%, perform grouped drug administration. Set 3 replicates for each group, and add 2 mL of liquid to each well. Add culture medium to each well in the blank control (BC) group, add culture medium containing 300 μM H2O2 to each well in the negative control (NC) group, add culture medium containing 0.05% VE and 300 μM H2O2 to each well in the positive control (PC) group, and add culture medium containing 0.1, 1, 5 ppm of EL-5 sample and H2O2 to each well in the sample group. After completion, place the 6-well plate in a CO2 incubator (37 °C, 5% CO2) and culture for 24 h.

[0063] (3)ROS content detection: After the induction ends, directly wash the cells 3 times with PBS, add 1 mL of 10 μM DCFH-DA probe to each well, and incubate in a CO2 incubator (37 °C, 5% CO2) for 30 min; discard the culture medium containing DCFH-DA, wash 3 times with PBS, digest the cells with trypsin (0.25%), wash the cells 1 time with PBS, add a certain amount of fresh PBS, perform flow cytometry detection, and calculate the down-regulation rate according to the formula: Down-regulation rate (%) = (Negative control group - Sample group) / Negative control group × 100%.

[0064] The experimental results are as Figure 5 shown. Compared with the blank control group BC, the ROS content was significantly up-regulated after H2O2-induced cells, indicating that the experimental model was successfully established (NC).

[0065] Compared with the negative control group (NC), the three concentrations of EL-5 samples could significantly down-regulate the ROS content (the inhibition rates were 4.35%, 13.82%, and 23.22% respectively), and showed a concentration-dependent relationship. Thus, under the experimental conditions of this study, the EL-5 sample has the ability to inhibit the ROS content in fibroblasts induced by H2O2, indicating that EL-5 has potential antioxidant properties.

[0066] Example 6 Fibroblast ATP Content Test Experiment Human primary dermal fibroblasts were amplified and cultured under the conditions of 5% CO2 and 37°C. When the cells grew to 70-80% confluence, they were digested with trypsin and seeded in a 96-well plate for 48 h. After changing to starvation medium for 18 h, grouping and drug administration were carried out. A blank control group (only adding medium) and a sample group (adding medium containing 0.1, 1, 5 ppm EL-5 samples) were set up, with 3 parallel samples in each group. After 24 h of culture, the cells were lysed after the experiment, and the ATP content of the cells was detected using CellTiter-Glo® reagent: reacting at room temperature in the dark for 10 min, and reading with a microplate reader (Luminescence). The test data were processed by combining with the ATP standard curve.

[0067] The experimental results are as Figure 6 shown. Compared with the NT group, the three concentrations of EL-5 samples could significantly promote the ATP content of fibroblasts (increased to 106.4%, 117.9%, 116.1% respectively). Thus, under the experimental conditions of this study, the EL-5 sample has the ability to increase the ATP content in fibroblasts, promote cell proliferation, and promote skin repair.

[0068] Example 7 IL-8 test experiment on HaCaT cells (1) Cell culture: Resuscitate the keratinocyte HaCaT. After the cell aggregation degree reaches 80%, seed the cells into a 24-well plate and incubate overnight in a CO2 incubator (37°C, 5% CO2).

[0069] (2) Ultraviolet induction: Remove the medium in the well plate, add the culture buffer DPBS. The model group, positive control group, and sample group received UVB irradiation with a total dose of 11 mJ / cm 2 At the same time, the blank control group was placed in the same environment (UVB radiation dose of 0 J / cm 2 ).

[0070] (3) Induction and sample addition: After the induction, remove the DPBS in the well plate. DMEM medium was added to each well of the blank control group and the model group, DMEM medium containing 1 μg / mL dexamethasone was added to each well of the positive control group, and DMEM medium containing 0.1, 1, 5 ppm EL-5 samples was added to each well of the sample group. After completion, the 24-well plate was placed in a CO2 incubator (37°C, 5% CO2) for 24 h.

[0071] (4) Determination of IL-8 content: Collect the cell culture medium, centrifuge at 3000 rpm for 3 min, and take the supernatant. Detect the IL-8 content according to the kit instructions, use the standard concentration as the horizontal axis and the OD value as the vertical axis, and use computer software to perform regression fitting to generate a standard curve. Determine the best fitting curve by regression analysis. Calculate the corresponding IL-8 content in the sample to be tested.

[0072] The experimental results are as follows Figure 7 Compared with the blank control group, the IL-8 content in HaCaT cells induced by UVB was significantly increased, indicating that the experimental model was successfully established.

[0073] Compared with the model group, the three concentrations of EL-5 (0.1, 1, 5 ppm) can significantly inhibit the secretion of IL-8, reducing it by 17%, 23% and 12% respectively, and the differences between the concentration groups are statistically significant. This shows that EL-5 can effectively antagonize the release of IL-8 inflammatory factors induced by UVB, suggesting that it plays an important role in regulating the inflammatory pathway related to skin photodamage.

[0074] The above is an explanation of the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A bioactive peptide, characterized in that: Its amino acid sequence is as shown in SEQ ID No.1, or it is a derivative with the same biological activity obtained by hydroxylating, carboxylating, carbonylating, methylating, acetylating, phosphorylating, esterifying or glycosylating the amino acid side chain group, amino terminal or carboxyl terminal of the SEQ ID No.1 sequence.

2. A pharmaceutical composition, characterized in that The invention comprises the bioactive peptide according to claim 1.

3. The pharmaceutical composition according to claim 2, characterized in that The invention also contains a pharmaceutically acceptable carrier and / or a fermentation product of Saccharomyces cerevisiae with a preservation number of CGMCC No.19732.

4. The pharmaceutical composition according to claim 3, wherein the pharmaceutically acceptable carrier is selected from any one or any combination of isotonic saline, ethanol, phosphate buffered saline, sorbitol, mannitol, starch, acacia gum, gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl pyrrolidone, cellulose and water; and the fermentation product is selected from any one or any combination of fermentation broth, fermentation filtrate, concentrated solution of fermentation broth or fermentation filtrate, and dry powder of fermentation broth or fermentation filtrate.

5. The pharmaceutical composition of any one of claims 2 to 4, wherein the dosage form comprises any one of an oral dosage form, a parenteral dosage form, a buccal dosage form, a sublingual dosage form, a nasal dosage form, an inhaler, a nebulizer, a topical dosage form, a transdermal dosage form and a suppository, or any combination thereof.

6. A daily chemical composition, characterized in that: The invention comprises the bioactive peptide according to claim 1.

7. The daily chemical composition according to claim 6, characterized in that: The invention also contains fermentation products of saccharomyces cerevisiae with the deposit number of CGMCC No.19732.

8. Use of the bioactive peptide according to claim 1, the pharmaceutical composition according to any one of claims 2 to 5, or the daily chemical composition according to any one of claims 6 to 7 in the preparation of drugs, health products or foods with anti-aging effects.

9. Use of the bioactive peptide according to claim 1, the pharmaceutical composition according to any one of claims 2 to 5, or the daily chemical composition according to any one of claims 6 to 7 in the preparation of drugs, health products or foods having antioxidant and / or skin cell proliferation promoting effects and / or type I collagen content increasing effects.

10. Use of the bioactive peptide according to claim 1, the pharmaceutical composition according to any one of claims 2 to 5, or the daily chemical composition according to any one of claims 6 to 7 in the preparation of skin care products or cosmetics with anti-aging, and / or sun protection, and / or anti-photoaging effects.

11. Use of the bioactive peptide according to claim 1, the pharmaceutical composition according to any one of claims 2 to 5, or the daily chemical composition according to any one of claims 6 to 7 in the preparation of skin care products or cosmetics having antioxidant effects, and / or promoting skin cell proliferation, and / or increasing the content of type I collagen, and / or promoting the production and secretion of type I collagen, and / or reducing inflammatory response effects.

12. A skin care product or cosmetic, characterized in that: Contains the bioactive peptide according to claim 1, the pharmaceutical composition according to any one of claims 2 to 5, or the daily chemical composition according to any one of claims 6 to 7.

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