Spirulina phycocyanin polypeptide with moisturizing, repairing and anti-aging effects and application thereof

Abstract

The invention relates to spirulina phycocyanin polypeptide with moisturizing, repairing and anti-aging effects and application of the spirulina phycocyanin polypeptide, and belongs to the technical field of active polypeptide. The active peptide with the moisturizing, repairing and anti-aging effects and a specific sequence is screened from enzymolysis products of spirulina phycocyanin, and the active peptide is polypeptide with the amino acid sequence being NSPL or GHP. The natural, safe and efficient RXR alpha receptor binding agent is prepared, the application of small-molecule polypeptide in the field of cosmetics is expanded, and the RXR alpha receptor binding agent has good application prospects and excellent economic value.

Description

Technical Field

[0001] The invention relates to a spirulina phycocyanin polypeptide with moisturizing, repairing and anti-aging effects and application thereof, belonging to the technical field of active polypeptides. Background Art

[0002] Phycobiliprotein is a natural light-harvesting pigment protein found in microalgae, cyanobacteria and some eukaryotic algae. Phycobiliprotein located on the thylakoid membrane of algae is stacked and assembled into phycobilisomes according to a certain structure. According to the type of chromophores and absorption spectra they contain, phycobiliprotein can be divided into four categories: phycocyanin (PC), phycoerythrin (PE), allophycocyanin (APC) and phycoerythrocyanin (PEC). Phycobiliprotein is water-soluble, non-toxic, brightly colored, and can emit bright fluorescence. It has multiple biological activities such as anti-oxidation, anti-radiation, anti-inflammatory, anti-viral, anti-tumor, immune regulation and nerve protection, and is widely used in food, cosmetics, pharmaceuticals and other fields.

[0003] Phycocyanin active peptides have a small molecular weight, are easily absorbed, and have a clear mechanism of action, so they can effectively penetrate deep into the skin and exert their biological effects. They can be used as additives for high-end products in the cosmetics field to enhance the efficacy and competitiveness of products. However, due to the complex screening process and high cost, they are rarely used in the cosmetics field. Summary of the invention

[0004] The present invention provides a spirulina phycocyanin polypeptide with moisturizing, repairing and anti-aging effects. The present invention uses spirulina phycocyanin as a parent protein, adopts alkaline protease and papain combined enzymatic hydrolysis to prepare a novel phycocyanin-derived beauty peptide, predicts the physicochemical properties, toxicity, antioxidant activity, anti-aging activity and the like of the obtained polypeptide by bioinformatics methods such as molecular docking technology, screens an ideal novel phycobiliprotein-derived beauty peptide from the peptide, synthesizes the peptide by chemical solid phase synthesis, and systematically evaluates the moisturizing, repairing and anti-aging effects of the novel beauty peptide by in vitro experiments. The results show that the polypeptide screened by the present invention has moisturizing, repairing and anti-aging effects, and can be used for the research and development of cosmetics, functional foods and health products.

[0005] The technical solution of the present invention is as follows:

[0006] A polypeptide whose amino acid sequence is NSPL, i.e., Asn-Ser-Pro-Leu.

[0007] A polypeptide whose amino acid sequence is GHP, i.e. Gly-His-Pro.

[0008] The above two polypeptides NSPL and GHP are bioactive peptides derived from Spirulina phycocyanin. By optimizing the enzymatic hydrolysis conditions, the enzymatic hydrolysis products with higher activity were screened, and then separated and purified by ultrafiltration and high performance liquid chromatography (HPLC). The structure of the phycocyanin-derived active peptide components was analyzed by LC-MS / MS, and then bioinformatics methods were used to predict the physicochemical properties, potential biological activity, toxicity, etc. of the polypeptides, and the bioactive polypeptides were screened out.

[0009] A composition comprises the above-mentioned polypeptide with the amino acid sequence of NSPL and the polypeptide with the amino acid sequence of GHP.

[0010] Application of the above polypeptide or composition in the preparation of products with moisturizing effect.

[0011] The use of the above polypeptide or composition in the preparation of products with repairing effects.

[0012] Use of the above polypeptide or composition in preparing products with anti-aging effects.

[0013] Preferably according to the present invention, the polypeptide or composition achieves the anti-aging effect by binding to the RXRα receptor and / or promoting the expression of Collagen I.

[0014] In the present invention, the above-mentioned products with moisturizing, repairing and anti-aging effects include: any one or more of cosmetics, skin care products, medical beauty products, medicines and health care products. The skin care products include: any one or more of facial cleansers, lotions, toners, moisturizing lotions, eye essences, moisturizing creams, essences, skin bases, sunscreens and facial masks. The cosmetics include: any one or more of liquid foundation and isolation cream. The medical beauty products include: any one or more of medical beauty masks, microneedles and water light needles. The dosage form of the medicine is selected from: any one or more of injections, tablets, capsules, granules, ointments, controlled release agents and aerosols.

[0015] Application of the above polypeptide or composition in the preparation of RXRα receptor binding products.

[0016] A skin product comprising the above polypeptide or composition.

[0017] In the present invention, the skin products include any one or more of cosmetics, skin care products and medical beauty products. The skin care products include any one or more of cleansers, lotions, toners, moisturizing lotions, eye essences, moisturizing creams, essences, skin bases, sunscreens and masks. The cosmetics include any one or more of foundation and isolation cream. The medical beauty products include any one or more of medical beauty masks, microneedles and water light needles.

[0018] The preparation method of the above polypeptide or composition is to obtain it from Spirulina phycocyanin by enzymatic hydrolysis and separation, or to obtain it by chemical synthesis according to the amino acid sequence.

[0019] Beneficial effects:

[0020] The present invention screens out active peptides with specific sequences having moisturizing, repairing and anti-aging effects from the enzymatic hydrolysis products of spirulina phycocyanin, wherein the active peptides are polypeptides with amino acid sequences of NSPL or GHP. The present invention prepares a natural, safe and efficient RXRα receptor binder, expands the application of small molecule polypeptides in the field of cosmetics, and has good application prospects and excellent economic value. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 The HPLC spectrum of the peptide sample containing NSPL.

[0022] Figure 2 This is the MS spectrum of the peptide NSPL.

[0023] Figure 3 The HPLC spectrum of the polypeptide sample containing GHP.

[0024] Figure 4 This is the MS spectrum of the peptide GHP.

[0025] Figure 5 This is the simulated docking interaction diagram between peptide NSPL and RXRα receptor.

[0026] Figure 6 This is the simulated docking interaction diagram between peptide GHP and RXRα receptor. DETAILED DESCRIPTION

[0027] The technical scheme of the present invention will be further described by combining specific embodiments. However, it should be understood that the embodiments described in the present invention are only exemplary and do not constitute any limitation on the scope of the present invention. The reagents and materials involved in the embodiments are all common commercial products unless otherwise specified. The biological materials involved in the embodiments can be obtained from common commercial sources unless otherwise specified.

[0028] Example 1: Preparation of Spirulina Phycocyanin Peptide

[0029] The preparation method of Spirulina phycocyanin peptide comprises the following steps:

[0030] 1) Spirulina phycocyanin powder was obtained from Shanghai Kefan Chemical Co., Ltd. The spirulina phycocyanin powder was dissolved and the pH of the spirulina phycocyanin solution was adjusted to 9. Then, 2×10 5U / g of composite protease (alkaline protease and papain activity ratio of 1:1) was added, mixed and placed in a shaker for oscillation enzymolysis at a shaker speed of 150 rpm, enzymolysis time of 5 h, and enzymolysis temperature of 40 ° C. After the enzymolysis, the enzymolysis was terminated by boiling water bath for 10 min to inactivate the enzyme;

[0031] 2) The enzymatic hydrolyzate was centrifuged at 12000 rpm for 20 min at 4°C, the supernatant was collected, and the precipitate was discarded; the supernatant was the Spirulina phycocyanin peptide solution; then a 3 kDa ultrafiltration membrane was used for ultrafiltration to obtain a peptide solution with a molecular weight of <3 kDa, which was freeze-dried to obtain a Spirulina phycocyanin peptide freeze-dried powder.

[0032] Example 2: Structural Identification of Spirulina Phycocyanin Peptide

[0033] The lyophilized powder of the spirulina phycocyanin peptide obtained in Example 1 was re-dissolved with ultrapure water, filtered through a 0.22 μm water filter membrane, and samples were collected according to the peak time by liquid chromatography to perform preliminary separation of the polypeptide samples, and then the sequence structure of the spirulina phycocyanin peptide was analyzed and identified by ultra-high performance liquid chromatography tandem mass spectrometry. In ultra-high performance liquid chromatography, mobile phase A (0.1% formic acid) aqueous solution, mobile phase B (0.1% formic acid) acetonitrile solution, elution time was 60min, gradient elution conditions were shown in Table 1, injection volume was 5 μL, and flow rate was set to 300 μL / min. Tandem mass spectrometry adopted positive ion mode, secondary mass spectrometry analysis, and full MS scan of 100-2000 m / z was performed at a resolution of 120000, and specific mass spectrometry parameter settings were shown in Table 2.

[0034] Table 1. ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY ELUTATION CONDITIONS

[0035]

[0036]

[0037] Table 2. Tandem mass spectrometry analysis conditions

[0038]

[0039] Mass spectrometry data interpretation:

[0040] The original mass spectrometry files were converted into MGF format files using MM File Conversion software, and then the MASCOT mass spectrometry data analysis platform was used to retrieve protein data in the uniport database. The specific search parameters are shown in Table 3.

[0041] Table 3. Mascot search parameters

[0042]

[0043] Note: Search the comparison database http: / / www.uniprot.org / taxonomy / 8139,

[0044] Search software http: / / www.matrixscience.com / .

[0045] Example 3: Bioinformatics prediction of Spirulina phycocyanin peptides

[0046] 1) Prediction of potential biological activity and toxicity of Spirulina phycocyanin peptides

[0047] The potential biological activity of the obtained peptide sequences was analyzed using the PeptideRanker online platform (http: / / distilldeep.ucd). The peptides were ranked according to their predicted probability of biological activity. The preset threshold of the PeptideRanker prediction model was 0.5, and any peptide with a threshold exceeding 0.5 was marked as having biological activity. In this study, a threshold of > 0.8 was selected as the screening condition for the biological activity of the peptides.

[0048] Toxicity and sensitization are also concerns in the research and development of peptides used in the field of skin care products. The ToxinPrep (https: / / webs.iiitd.edu.in / raghava / toxinpred / multi_submit.php) platform is used to predict the potential toxicity of structurally resolved peptide sequences based on the SVM (Swiss-Port) algorithm.

[0049] 2) Prediction of antioxidant activity of Spirulina phycocyanin peptides

[0050] The antioxidant activity of the non-toxic and potentially biologically active peptide sequences with PeptideRanker>0.8 was predicted using the AnOxPePred (https: / / services.healthtech.dtu.dk / service.phpAnOxPePred-1.0) platform, and the analysis model selected Peptide Mode, and the Minimum peptide length and Maximum peptide length were set to the peptide chain length of the predicted peptide. In this study, the prediction results of FRS score>0.5 or CHEL score>0.25 were selected as signs of peptides with antioxidant activity.

[0051] 3) Molecular docking experiment of Spirulina phycocyanin peptide and cellular RXRα receptor

[0052] The non-toxic peptide sequences with potential biological activity and antioxidant activity were used for docking with the cell RXRα receptor molecules. The peptide sequence was docked with the RXRα receptor sequence from the PDB database with the accession number PDB ID: P19793 using pepsite 2.0 (http: / / pepsite2.russelllab.org / ) and AutoDock 1.5.6 to simulate molecular docking experiments and calculate the docking score. The RXRα receptor target structure and the peptide sequence were docked using vina in the pyrx software (https: / / pyrx.sourceforge.io / ). The Affinity value (kcal / mol) represents the binding ability of the two. The lower the Affinity value, the more stable the binding of the ligand to the receptor.

[0053] Bioinformatics prediction results:

[0054] Through the above bioinformatics prediction experiment, two Spirulina phycocyanin peptides were screened out, namely: NSPL (Asn-Ser-Pro-Leu) and GHP (Gly-His-Pro). Among them, the HPLC spectrum of the peptide sample containing NSPL is as follows Figure 1 As shown, the MS spectrum of peptide NSPL is as follows Figure 2 As shown, the HPLC spectrum of the polypeptide sample containing GHP is as follows Figure 3 As shown, the MS spectrum of the peptide GHP is as follows Figure 4 shown.

[0055] Simulated docking interaction between peptide NSPL and RXRα receptor Figure 5 As shown, the simulated docking interaction between peptide GHP and RXRα receptor is shown in Figure 6As shown, the binding ability of polypeptides NSPL and GHP to RXRα receptor is shown in Table 4. From the above results, it can be seen that the polypeptide NSPL forms hydrogen bonds with ASP448, THE266, ASN262, THR445, and PRO446 of the RXRα receptor protein, and the binding is relatively tight; the polypeptide GHP forms hydrogen bonds with PRO446, THR445, THR266, VAL265, and GLY443 of the RXRα receptor protein, and the binding is relatively tight. Retinoic acid receptor (RARα) belongs to one of the nuclear receptor subfamily (RARα, RARβ and RARγ). After the polypeptide binds to the relevant receptors (RXR, RAR), it forms various dimers and binds to the corresponding retinoic acid response element with the assistance of activators (such as histone acetyltransferase). By regulating and controlling the activation and expression of messenger RNA in the cell nucleus, it regulates cell division and differentiation, stimulates keratinocyte proliferation, promotes the metabolism of epidermal keratinocytes, reduces the degradation of dermal collagen, and stimulates fibroblasts to produce more collagen, hyaluronic acid and other substances, thereby achieving an anti-aging effect.

[0056] Table 4. Binding ability of peptides to RXRα receptor

[0057]

[0058] It is generally believed that an affinity value less than -4.25 kcal / mol indicates a certain binding activity between the two, less than -5.0 kcal / mol indicates a good binding activity, and less than -7.0 kcal / mol indicates a strong binding activity.

[0059] 4) Chemical synthesis of Spirulina phycocyanin peptide

[0060] The above two polypeptide sequences were chemically synthesized using Fmoc amino acid solid phase synthesis technology. The chemical synthesis was completed by Nanjing Peptide Valley Biotechnology Co., Ltd. The purity was ≥95%, and the purity and molecular weight of the synthesized phycocyanin-derived active polypeptides identified by RP-HPLC and MS met the requirements.

[0061] Example 4: In vitro promotion of type I collagen synthesis experiment

[0062] The experiment used human skin fibroblasts (HSF cells) (batch number: Fb220309, provided by Guangdong Boxi Biotechnology Co., Ltd.) as a model cell line for research. The experimental steps are as follows:

[0063] 1) Cell inoculation: After thawing the cells, when the plating rate reaches about 60%, digest and resuspend the cells to obtain a cell suspension, inoculate the cell suspension into a 6-well plate at a volume of 2 mL / well, and incubate in a CO2 incubator (37°C, 5% CO2) overnight;

[0064] 2) Solution preparation: Prepare the working solution of the test substance according to the test group (Table 5);

[0065] Table 5. Test grouping scheme

[0066]

[0067] 3) Drug administration: According to the test grouping, when the cell plating rate in the 6-well plate reaches 40% to 60%, group drug administration is carried out, and 3 replicate wells are set up for each group; after discarding the old culture medium, 2 mL of culture medium is added to each well of the blank control group, 2 mL of culture medium containing the corresponding concentration of TGF-β1 is added to each well of the positive control group, and 2 mL of culture medium containing the corresponding concentration of the sample to be tested is added to each well of the sample group; after drug administration, the 6-well plate is placed in a CO2 incubator (37°C, 5% CO2) and cultured for 24 hours;

[0068] 4) Collect cells: After the culture is completed, discard the old solution, wash twice with PBS buffer, add 1 mL of RNAisoPlus to each well, pipette and lyse the cells, and collect the samples;

[0069] 5) ELISA detection: collect the supernatant and detect and analyze the type I collagen concentration in the supernatant according to the operating instructions of the ELISA kit;

[0070] 6) Calculation of the enhancement rate: Calculate the enhancement rate of type I collagen according to the following formula:

[0071]

[0072] 7) Statistical analysis of results: GraphPad Prism was used for graphing, and the results were expressed as Mean±SD. t-test was used for comparison among the groups. All statistical analyses were two-tailed. P<0.05 was considered to be significantly different, and P<0.01 was considered to be extremely significantly different. The results are shown in Table 6.

[0073] Table 6. CollagenⅠ content detection results

[0074] Experimental Grouping Average concentration (ng / mL) SD P-value Improvement rate (vs BC) BC 20.67 2 / / PC 34.46 2.35 0.002** 66.72％ Sample set 1 (NSPL) 48.37 2.27 0.000** 134.00％ Sample set 2 (GHP) 39.84 2.11 0.000** 92.74％

[0075] Note: When the t-test method was used for statistical analysis, the significance was indicated by *, P-value < 0.05 was indicated by *, and P-value < 0.01 was indicated by ** compared with the BC group.

[0076] As can be seen from Table 6: compared with the BC group, the content of Collagen Ι in the PC group increased significantly, indicating that the positive control of this test is effective; compared with the BC group, the content of Collagen Ι in the NSPL and GHP peptides at a concentration of 2.5 mg / mL increased significantly, with an increase rate of 134.00% and 92.74% respectively. Therefore, it is shown that the above two peptides can promote a significant increase in the content of Collagen I and achieve a firming and anti-aging effect.

[0077] Example 5: In vitro experiment on promoting AQP3 synthesis

[0078] The experiment used keratinocytes (lot number: Ep23082202, provided by Guangdong Boxi Biotechnology Co., Ltd.) as the model cell line for research. The experimental steps are as follows:

[0079] 1) Cell inoculation: After thawing the cells, when the plating rate reaches about 60%, digest and resuspend the cells to obtain a cell suspension, inoculate the cell suspension into a 24-well plate at a volume of 1 mL / well, and incubate in a CO2 incubator (37°C, 5% CO2) overnight;

[0080] 2) Solution preparation: Prepare the working solution of the test substance according to the test group (Table 7);

[0081] Table 7. Test grouping scheme

[0082]

[0083] 3) Administration: According to the test grouping, when the cell plating rate in the 24-well plate reaches 40% to 60%, group administration is carried out, and 3 replicate wells are set up for each group; after discarding the old culture medium, 1 mL of culture medium is added to each well of the blank control group, 1 mL of culture medium containing the corresponding concentration of CaCl2 is added to each well of the positive control group, and 1 mL of culture medium containing the corresponding concentration of the sample to be tested is added to each well of the sample group; after the administration is completed, the 24-well plate is placed in a CO2 incubator (37°C, 5% CO2) and cultured for 24 hours;

[0084] 4) Immunofluorescence detection: After the culture, the cells were fixed with 4% paraformaldehyde for 30 minutes before immunofluorescence detection. The cells were photographed and observed under a microscope. The images were collected and the relative IOD of the target signals in the images was analyzed using Ipwin32 image analysis software. The average value was calculated and the relative IOD / cell number average value was calculated based on the blank control group to characterize the relative expression of AQP3 protein.

[0085] 5) Calculation of improvement rate: Calculate the AQP3 improvement rate according to the following formula:

[0086]

[0087] 6) Statistical analysis of results: GraphPad Prism was used for graphing, and the results were expressed as Mean±SD. t-test was used for comparison among the groups. All statistical analyses were two-tailed. P<0.05 was considered to be significantly different, and P<0.01 was considered to be extremely significantly different. The results are shown in Table 8.

[0088] Table 8. AQP3 immunofluorescence analysis results

[0089] Experimental Grouping Relative IOD / average cell number SD P-value Improvement rate (vs BC) BC 1 0.12 / / PC 1.42 0.11 0.011* 42％ Sample set 1 (NSPL) 1.81 0.20 0.003** 81.00％ Sample set 2 (GHP) 1.66 0.24 0.013* 66.00％

[0090] Note: The integrated optical density (IOD) reflects the content of AQP3. When the t-test method is used for statistical analysis, the significance is indicated by *, P-value < 0.05 is indicated by *, and P-value < 0.01 is indicated by ** when compared with the BC group.

[0091] Aquaporin (AQP) is a type of transport protein related to the permeability of water molecules on the cell membrane. AQP generally selectively transports water molecules, but a few AQPs can also transport small molecules such as glycerol and urea. AQP3, which is mainly expressed in the skin, is a transport protein that transports small molecules such as water, glycerol and urea across the cell membrane. It is the most expressed aquaporin subtype in human skin. From the results of AQP3 immunofluorescence analysis in Table 8, it can be seen that compared with the BC group, the AQP3 content of the PC group increased significantly, indicating that the positive control of this test is effective; compared with the BC group, the AQP3 content of the polypeptides NSPL and GHP at a concentration of 2.5 mg / mL increased significantly, with an increase rate of 81.00% and 66.00%, respectively. Therefore, it is shown that the above two polypeptides can promote a significant increase in the AQP3 content and achieve a moisturizing effect.

[0092] Example 6: In vitro promotion of FLG and LOR synthesis experiment

[0093] The experiment used keratinocytes as a model cell line for research. The experimental steps are as follows:

[0094] 1) Cell inoculation: After thawing the cells, when the plating rate reaches about 60%, digest and resuspend the cells to obtain a cell suspension, inoculate the cell suspension into a 6-well plate at a volume of 2 mL / well, and incubate in a CO2 incubator (37°C, 5% CO2) overnight;

[0095] 2) Solution preparation: Prepare the working solution of the test substance according to the test group (Table 9);

[0096] Table 9. Test grouping scheme

[0097]

[0098] 3) Administration: According to the test grouping, when the cell plating rate in the 6-well plate reaches 40% to 60%, group administration is carried out, and 3 replicate wells are set up for each group; after discarding the old culture medium, 2 mL of culture medium is added to each well of the blank control group, 2 mL of culture medium containing the corresponding concentration of WY14643 is added to each well of the PC group, and 2 mL of culture medium containing the corresponding concentration of the sample to be tested is added to each well of the sample group; after the administration is completed, the 6-well plate is placed in a CO2 incubator (37°C, 5% CO2) and cultured for 24 hours;

[0099] 4) Collect cells: After the culture is completed, discard the old solution, wash twice with PBS buffer, add 1 mL of RNAisoPlus to each well, pipette and lyse the cells, and collect the samples;

[0100] 5) Gene expression detection: RNA was extracted and reverse transcribed into cDNA, and then fluorescence quantitative PCR was performed. -△△CT Methods The mRNA expression levels of FLG and LOR were calculated;

[0101] 6) Calculation of up-regulation rate: The up-regulation rate of mRNA expression of FLG and LOR was calculated according to the following formula:

[0102]

[0103] 7) Statistical analysis of results: GraphPad Prism was used for graphing, and the results were expressed as Mean±SD. t-test was used for comparison among the groups. All statistical analyses were two-tailed. P<0.05 was considered to be significantly different, and P<0.01 was considered to be extremely significantly different. The results are shown in Tables 10 and 11.

[0104] Table 10. FLG mRNA content detection results

[0105] Experimental Grouping Mean SD P-value Increase rate (vs BC) BC 1.00 0.07 / / PC 1.23 0.14 0.06* 23.00％ Sample set 1 (NSPL) 1.91 0.13 0.000** 91.00％ Sample set 2 (GHP) 1.79 0.09 0.000** 79.00％

[0106] Note: Use 2- △△CT Methods The results were calculated and the mRNA content of the BC group was normalized when the t-test method was used for statistical analysis. Compared with the BC group, the significance was indicated by *, P-value < 0.05 was indicated by *, and P-value < 0.01 was indicated by **.

[0107] Table 11. Results of detection of mRNA content of LOR

[0108] Group Name Mean SD P-value Increase rate (vs BC) BC 1.00 0.05 / / PC 1.22 0.12 0.04* 22.00％ Sample set 1 (NSPL) 1.85 0.10 0.000** 85.00％ Sample set 2 (GHP) 1.74 0.13 0.000** 74.00％

[0109] Note: Use 2- △△CTMethods The results were calculated and the mRNA content of the BC group was normalized when the t-test method was used for statistical analysis. Compared with the BC group, the significance was indicated by *, P-value < 0.05 was indicated by *, and P-value < 0.01 was indicated by **.

[0110] Filaggrin (FLG) and loricrin (LOR) are proteins closely related to the skin barrier function. It can be seen from the gene detection results in Tables 10 and 11 that compared with the BC group, the mRNA content of FLG and LOR in the PC group increased significantly, indicating that the positive control test was effective; compared with the BC group, the mRNA content of FLG of the polypeptide NSPL and GHP at a concentration of 2.5 mg / mL increased significantly, with an increase rate of 91.00% and 79.00%, respectively, and the mRNA content of LOR increased significantly, with an increase rate of 85.00% and 74.00%, respectively. Therefore, it is shown that the above two polypeptides can increase the content of FLG and LOR at this concentration and have the effect of repairing the skin barrier.

Claims

1. A polypeptide, characterized in that Its amino acid sequence is NSPL, namely Asn-Ser-Pro-Leu.

2. A polypeptide, characterized in that Its amino acid sequence is GHP, i.e. Gly-His-Pro.

3. A composition, characterized in that It comprises the polypeptide whose amino acid sequence is NSPL as described in claim 1 and the polypeptide whose amino acid sequence is GHP as described in claim 2.

4. Use of the polypeptide according to claim 1 or 2 or the composition according to claim 3 in preparing a product with moisturizing effect.

5. Use of the polypeptide according to claim 1 or 2 or the composition according to claim 3 in preparing a product with repairing effect.

6. Use of the polypeptide according to claim 1 or 2 or the composition according to claim 3 in the preparation of a product with anti-aging effect.

7. The use according to claim 6, characterized in that: The polypeptide or composition achieves the anti-aging effect by binding to the RXRα receptor and / or promoting the expression of Collagen I.

8. The use according to any one of claims 4 to 6, characterized in that: The products include: any one or more of cosmetics, skin care products, medical beauty products, medicines and health care products; Preferably, the skin care products include: any one or more of: facial cleanser, lotion, toner, moisturizing lotion, eye essence, moisturizing cream, essence, skin base, sunscreen and facial mask; Preferably, the cosmetics include: any one or more of liquid foundation and isolation cream; Preferably, the medical beauty products include: any one or more of medical beauty masks, microneedles and water light needles; Preferably, the dosage form of the drug is selected from any one or more of injection, tablet, capsule, granule, ointment, controlled release agent and aerosol.

9. Use of the polypeptide according to claim 1 or 2 or the composition according to claim 3 in the preparation of a RXRα receptor binding product.

10. A skin product comprising the polypeptide of claim 1 or 2 or the composition of claim 3; Preferably, the skin product comprises: Any one or more of cosmetics, skin care products and medical beauty products; Further preferably, the skin care products include: any one or more of: facial cleanser, lotion, toner, moisturizing lotion, eye essence, moisturizing cream, essence, skin base, sunscreen and facial mask; Further preferably, the cosmetics include: any one or more of liquid foundation and isolation cream; Further preferably, the medical beauty product includes: any one or more of medical beauty masks, microneedles and hyaluronic acid needles.

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