A spirulina phycocyanin polypeptide for relieving inflammatory response, and its preparation method and application

Abstract

The present application belongs to the field of active peptide technology, and specifically relates to a Spirulina phycocyanin polypeptide for relieving inflammatory reactions, and its preparation method and application. The Spirulina phycocyanin polypeptide sequence is shown in SEQ ID NO.1. The polypeptide sequence structure is obtained through enzymatic hydrolysis, separation, and structural identification. The anti-inflammatory ability of the active peptide is verified by an in vitro anti-inflammatory experiment on macrophages.

Description

Technical Field

[0001] The present application relates to the technical field of active peptides, and more specifically, to a Spirulina phycocyanin polypeptide for relieving inflammatory responses, and a preparation method and application thereof. Background Art

[0002] Inflammation is an animal's defensive response to adverse stimuli. It also underlies the development of many diseases and can easily cause damage to the body. Clinically, glucocorticoids, with their powerful and rapid anti-inflammatory effects, are often used to treat inflammation, with dexamethasone being the most commonly used. However, reports indicate that long-term use of glucocorticoids may lead to adverse reactions such as anaphylactic shock, mental disturbances, and allergic rashes. Therefore, the development and research of special medical foods with anti-inflammatory properties that are easily absorbed by the human body and have no toxic side effects is imperative. Summary of the Invention

[0003] The present application is provided to address the above-mentioned deficiencies in the prior art. A Spirulina phycocyanin polypeptide that can relieve inflammatory reactions, as well as a preparation method and application thereof, is needed, and a small molecule peptide with anti-inflammatory effects is provided.

[0004] In a first aspect of the present application, a Spirulina phycocyanin polypeptide for relieving inflammatory response is provided, wherein the Spirulina phycocyanin polypeptide sequence is shown as SEQ ID NO. 1.

[0005] In a second aspect of the present application, a method for preparing the Spirulina phycocyanin polypeptide according to any embodiment of the present application is provided, the preparation method comprising: soaking Spirulina platensis in water to obtain a mixture A, freezing the mixture A, and then thawing the mixture A; centrifuging the thawed mixture A, and taking the supernatant as a crude phycocyanin extract; adjusting the crude phycocyanin extract to be alkaline, adding alkaline protease for enzymatic hydrolysis to obtain an enzymatic hydrolyzate; after centrifuging the enzymatic hydrolyzate, taking the supernatant, filtering and drying the supernatant to obtain a freeze-dried powder; and separating and analyzing the freeze-dried powder using ultra-performance liquid chromatography to obtain the Spirulina phycocyanin polypeptide shown in SEQ ID NO. 1.

[0006] The third aspect of the present application provides a use of the Spirulina phycocyanin polypeptide described in any embodiment of the present application in the preparation of an anti-inflammatory product.

[0007] The fourth aspect of the present application provides a use of the Spirulina phycocyanin polypeptide described in any embodiment of the present application in the preparation of an antioxidant product.

[0008] In a fifth aspect of the present application, a drug is provided, wherein the active ingredient of the drug comprises at least the Spirulina phycocyanin polypeptide described in any embodiment of the present application.

[0009] The Spirulina phycocyanin polypeptides for soothing inflammatory reactions, as well as their preparation methods and applications, provided in various embodiments of the present application, have been verified to have anti-inflammatory capabilities through in vitro anti-inflammatory experiments on macrophages. Therefore, the novel active peptide MGHP can be used as a functional ingredient in cosmetics and medicines with anti-inflammatory effects, and has good application prospects. Compared with dexamethasone, it can avoid adverse reactions such as anaphylactic shock, mental disorders, and allergic rashes caused by long-term use. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] In the drawings, which are not necessarily drawn to scale, the same reference numerals may describe similar components in different views. The same reference numerals with letter suffixes or different letter suffixes may represent different instances of similar components. The accompanying drawings generally illustrate various embodiments by way of example and not limitation, and together with the description and claims, serve to illustrate the claimed embodiments. Where appropriate, the same reference numerals are used throughout the drawings to refer to the same or similar parts. Such embodiments are illustrative and are not intended to be exhaustive or exclusive of the present apparatus or method.

[0011] Figure 1 shows an HPLC spectrum of a polypeptide sample containing MGHP according to an embodiment of the present application;

[0012] Figure 2 Shown is the MS spectrum of the polypeptide MGHP according to an example of the present application. DETAILED DESCRIPTION

[0013] In order to enable those skilled in the art to better understand the technical solution of the present application, the present application is described in detail below in conjunction with the accompanying drawings and specific embodiments. The embodiments of the present application are further described in detail below in conjunction with the accompanying drawings and specific embodiments, but are not intended to limit the present application.

[0014] The terms "first," "second," and similar terms used in this application do not denote any order, quantity, or importance, but are used only to distinguish. The terms "include," "comprise," and similar terms mean that the elements preceding the term include the elements listed after the term, and do not exclude the possibility of also including other elements.

[0015] According to the embodiments of the present application, a Spirulina phycocyanin polypeptide is provided for alleviating inflammatory responses. The Spirulina phycocyanin polypeptide sequence is shown in SEQ ID NO. 1. The amino acid sequence of the Spirulina phycocyanin polypeptide is MGHP, or Met-Gly-His-Pro. Peptides are generally more active than their parent proteins, and their molecular weight is smaller than that of proteins, making them easier for human absorption.

[0016] According to an embodiment of the present application, a method for preparing the Spirulina phycocyanin polypeptide according to any embodiment of the present application is provided, the preparation method comprising: soaking Spirulina platensis in water to obtain a mixture A, freezing the mixture A, and then thawing the mixture A; centrifuging the thawed mixture A, and taking the supernatant as a crude phycocyanin extract; adjusting the crude phycocyanin extract to be alkaline, adding alkaline protease for enzymatic hydrolysis to obtain an enzymatic hydrolyzate; after centrifuging the enzymatic hydrolyzate, taking the supernatant, filtering and drying the supernatant to obtain a freeze-dried powder; and separating and analyzing the freeze-dried powder using ultra-performance liquid chromatography-tandem mass spectrometry to obtain the Spirulina phycocyanin polypeptide shown in SEQ ID NO. 1.

[0017] The supernatant obtained after centrifugation of the thawed mixture A contains C-phycocyanin and allophycocyanin.

[0018] In some embodiments, the preparation method includes: soaking Spirulina platensis in water, with a volume ratio of Spirulina platensis to water being 1:18-22; freezing at a temperature of -18 to -22°C for 20-26 hours; and centrifuging the thawed mixture A at a temperature of 3-5°C.

[0019] In some embodiments, the conditions for centrifuging the melted mixture A include: a rotation speed of 5000-7000 rpm and a centrifugation time of 5-30 min. Specifically, the rotation speed may be 6000-6500 rpm and the centrifugation time may be 10-20 min.

[0020] In some embodiments, the melting temperature is 30-45° C. Specifically, the melting temperature may be 35-40° C.

[0021] In some embodiments, the pH of the phycocyanin crude extract is adjusted to be alkaline at 7.5-8.5; the activity of the alkaline protease added is (4-6)×10 5 ; The enzymatic hydrolysis time is 3-5h, the enzymatic hydrolysis temperature is 35-45℃; the temperature of the enzymatic hydrolyzate centrifugation is 3-5℃.

[0022] In some embodiments, the supernatant is filtered and dried to obtain a lyophilized powder, which is then ultrafiltered using a 3 kDa ultrafiltration membrane to obtain a peptide solution with a molecular weight of <3 kDa.

[0023] In some embodiments, the method of analyzing using ultra-high performance liquid chromatography-tandem mass spectrometry includes: filtering the lyophilized powder using an aqueous filter membrane, performing gradient elution on the filtrate through ultra-high performance liquid chromatography, and then analyzing it using tandem mass spectrometry; the tandem mass spectrometry uses a positive ion mode.

[0024] The freeze-dried powder is dissolved in water and then filtered using a water filter membrane.

[0025] In some embodiments, the pore size of the water filter membrane is 0.22 microns.

[0026] In some embodiments, the mobile phase A of the ultra-high performance liquid chromatography gradient elution is a 0.1% by mass formic acid aqueous solution, and the mobile phase B is an acetonitrile solution containing 0.1% by mass formic acid; the elution time is 55-65 min.

[0027] According to the embodiments of the present application, there is provided a use of the Spirulina phycocyanin polypeptide described in any embodiment of the present application in the preparation of an anti-inflammatory product.

[0028] The product can be a medicine, cosmetic or experimental reagent, etc., and the experimental reagent is used for basic research.

[0029] Furthermore, when the product is a drug, the drug can be used to prevent or treat diseases related to inflammation through its anti-inflammatory effect.

[0030] The pharmaceutical composition also includes pharmaceutically necessary carriers, which, of course, should be dosed in a manner that is harmless to the subject. These include, but are not limited to, buffers, antioxidants, preservatives, bactericides, proteins, hydrophilic polymers, amino acids, monosaccharides, disaccharides, and other carbohydrates, chelating agents, surfactants, salt-forming counterions, and metal complexes. Furthermore, the composition can be formulated into oral formulations, topical formulations, suppositories, and sterile injectable solutions in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, and sprays, according to conventional methods.

[0031] In some embodiments, the anti-inflammatory effect is manifested by inhibiting the secretion of macrophage inflammatory factors and inflammatory mediators, including IL-1B and TNF-a, and inflammatory mediators including PGE2.

[0032] According to the embodiments of the present application, there is provided a use of the Spirulina phycocyanin polypeptide described in any embodiment of the present application in the preparation of a product with antioxidant efficacy.

[0033] The product can be a medicine, cosmetic or experimental reagent, etc., and the experimental reagent is used for basic research.

[0034] According to an embodiment of the present application, a drug is provided, wherein the active ingredient of the drug comprises at least the Spirulina phycocyanin polypeptide described in any embodiment of the present application.

[0035] Example 1: Extraction of Spirulina Phycocyanin

[0036] Disrupt Spirulina using the freeze-thaw method: Place Spirulina powder and Polysiphonium into two 500ml reagent bottles. Add distilled water at a 1:20 (w / v) ratio to each. Soak overnight, freeze at -20°C for 24 hours, and then thaw in a 37°C water bath. Repeat this process three times. Centrifuge at 6000 rpm for 15 minutes at 4°C. The supernatant is the crude phycobiliprotein extract (containing C-phycocyanin and allophycocyanin).

[0037] Example 2: Preparation of Spirulina Phycocyanin Peptide

[0038] 1) Adjust the pH of the crude extract of Spirulina phycocyanin prepared in Example 1 to 8, and then add 5×10 5 Mix 1000 U / g of alkaline protease and place in a shaker for enzymatic hydrolysis at 100-200 rpm for 4 h at 40°C. Boil the hydrolyzate in boiling water for 10 min to inactivate the enzyme and terminate the enzymatic hydrolysis.

[0039] 2) Centrifuge the enzymatic hydrolyzate at 10,000-12,000 rpm for 15 minutes at 4°C. Collect the supernatant and discard the precipitate. The supernatant is the Spirulina phycocyanin peptide solution. Ultrafiltration is then performed using a 3 kDa ultrafiltration membrane to obtain a peptide solution with a molecular weight <3 kDa. Freeze-dry the solution to obtain a Spirulina phycocyanin peptide lyophilized powder.

[0040] Example 3: Structural Identification of Spirulina Phycocyanin Peptide

[0041] The lyophilized powder of Spirulina phycocyanin peptide obtained in Example 2 was re-dissolved in ultrapure water and filtered through a 0.22-μm aqueous filter membrane. Samples were collected based on peak elution time via liquid chromatography for peptide separation. The sequence structure of the Spirulina phycocyanin peptide was then analyzed and identified using ultra-high performance liquid chromatography-tandem mass spectrometry. The ultra-high performance liquid chromatography (ULHPLC-MS / MS) employed mobile phase A (0.1% formic acid in water) and mobile phase B (0.1% formic acid in acetonitrile). The elution time was 60 minutes, and the gradient elution conditions were as shown in Table 1. The injection volume was 5 μL, and the flow rate was set at 300 μL / min.

[0042] Table 1 Ultra-high performance liquid chromatography elution conditions

[0043]

[0044] The peptide samples separated by ultra-high performance liquid chromatography were analyzed by mass spectrometry. Tandem mass spectrometry was performed in positive ion mode, and secondary mass spectrometry was used for analysis. A full MS scan of 100-2000 m / z was performed at a resolution of 120,000. The specific mass spectrometry parameter settings are shown in Table 2.

[0045] Table 2 Tandem mass spectrometry analysis conditions

[0046]

[0047] Mass spectrometry data interpretation:

[0048] The original mass spectrometry files were converted into MGF format files using MM File Conversion software, and then the protein data in the uniport database were retrieved using the MASCOT mass spectrometry data analysis platform. The specific search parameters are shown in Table 3.

[0049] Table 3 Mascot search parameters

[0050]

[0051] Note: Search the comparison database http: / / www.uniprot.org / taxonomy / 8139,

[0052] Search software: http: / / www.matrixscience.com / .

[0053] Example 4: Bioinformatics prediction of Spirulina phycocyanin peptides

[0054] 1) Prediction of potential biological activity and toxicity of Spirulina phycocyanin peptides

[0055] The potential biological activities of the multiple polypeptide sequences obtained in Example 3 were analyzed using the PeptideRanker online platform (http: / / distilldeep.ucd). Peptides were ranked based on their predicted probability of biological activity. The PeptideRanker prediction model was pre-set at a threshold of 0.5; any polypeptide exceeding this threshold was labeled as biologically active. In this application, a threshold > 0.8 was selected as the screening criterion for peptides to be biologically active.

[0056] Toxicity and sensitization are also concerns in the development of peptides for skincare applications. Using the ToxinPrep (https: / / webs.iiitd.edu.in / raghava / toxinpred / multi_submit.php) platform and the SVM (Swiss-Port) algorithm, we predict the potential toxicity of peptide sequences with structural analysis.

[0057] The results showed that the multiple polypeptides separated in Example 3 were non-toxic.

[0058] 2) Prediction of antioxidant activity of Spirulina phycocyanin peptides

[0059] For the multiple peptides screened in Example 3 that were non-toxic and had a potential biological activity PeptideRanker score of >0.8, antioxidant activity prediction was performed using the AnOxPePred (https: / / services.healthtech.dtu.dk / service.php AnOxPePred-1.0) platform. The Peptide Mode analysis model was selected, and the Minimum peptide length and Maximum peptide length were both set to the peptide chain length of the predicted peptide. This application selected a prediction result with an FRS score >0.5 or a CHEL score >0.25 as a marker of peptide antioxidant activity.

[0060] The results showed that only some of the multiple peptides isolated from 3 had antioxidant activity.

[0061] 3) Prediction of anti-inflammatory and immune activities of Spirulina phycocyanin peptides

[0062] During an inflammatory response, cells release a variety of inflammatory mediators, such as interleukins, tumor necrosis factor (TNF), prostaglandins, and leukotrienes. These mediators can cause vasodilation and increased permeability, thereby exacerbating inflammation. Anti-inflammatory drugs, such as cyclooxygenase inhibitors, can inhibit the production of these mediators, thereby alleviating inflammation. The IL-1 and TNF-β secretion inhibitory peptide prediction module of the PHI platform was used to predict and analyze anti-inflammatory peptides.

[0063] Bioinformatics was used to predict the anti-inflammatory and immune activities of the peptides with antioxidant activity after the above 2) screening, and finally a peptide with good antioxidant and anti-inflammatory properties, MGHP, was screened out. Its score was 0.81, its molecular weight was 440.57, it could significantly inhibit the biological activities of IL-1 and TNF-β, and it had not appeared in the peptide database. Therefore, it can be used as a new anti-inflammatory peptide for subsequent research, and then its efficacy can be further verified through macrophage anti-inflammatory experiments.

[0064] 4) Chemical synthesis of Spirulina phycocyanin peptide

[0065] The peptide sequence of MGHP finally selected in Example 3 was 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%. The purity and molecular weight of the synthesized phycocyanin-derived active peptide were determined by RP-HPLC and MS. The HPLC spectrum of MGHP is shown in Figure 2. Figure 1 As shown, through Figure 1 It can be seen that the detection peak appears during the elution process from 6 minutes to 8 minutes. The MS spectrum of MGHP is as follows Figure 2 As shown, through Figure 2 The mass spectrum peaks can be found at 441.5-442.57.

[0066] Example 5: Cellular anti-inflammatory experiment

[0067] This experiment was divided into two parts: the first part was based on macrophages, and cytotoxicity testing was carried out to determine the dosage concentration of the chemically synthesized polypeptide MGHP in Example 4 on macrophages; the second part was to stimulate macrophages with lipopolysaccharide (LPS) and evaluate the anti-inflammatory efficacy of the test samples by detecting changes in the levels of inflammatory factors (IL-1β, TNF-α) and inflammatory mediators (PGE2).

[0068] The cells used in this test were macrophages, batch number: 210622-1, provided by Guangdong Boxi Biotechnology Co., Ltd.

[0069] Cytotoxicity test:

[0070] 1) Cell seeding: After thawing cells, when the plating rate reaches approximately 60%, seed the cells into a 96-well plate and incubate overnight in a CO2 incubator (37°C, 5% CO2).

[0071] 2) Experimental Grouping: The experiment was divided into a zero adjustment group, a solvent control group, a positive control group, and a sample group. Within the sample group, each sample was set up with 8 concentration gradients, and 3 replicate wells were set up under each concentration gradient.

[0072] 3) Solution preparation: Prepare sample working solutions of different concentrations according to the test concentration setting table (Table 4).

[0073] 4) Dosing: Dosing was performed when the cell plating rate in the 96-well plate reached 50%-60%. For the solvent control group (corresponding to the control in Table 4, 200 μL of DMEM (Gibco) culture medium was added to each well); for the positive control group (corresponding to the PC in Table 4, 200 μL of culture medium containing 10% DMSO was added to each well); for the sample group, 200 μL of culture medium containing the corresponding concentration of MGHP peptide and DMEM was added to each well; for the zero adjustment group, no cells were seeded, and only 200 mL of cell culture medium was added. After dosing, the 96-well plate was placed in a CO2 incubator (37°C, 5% CO2) for 24 hours.

[0074] 5) Detection: After incubating the cells for 24 h, discard the supernatant and add MTT working solution (0.5 mg / mL). Incubate at 37°C in the dark for 4 h. After incubation, discard the supernatant and add 150 μL of DMSO to each well. Read the OD value at 490 nm.

[0075] Calculation of relative cell viability:

[0076] Relative cell viability (%) .

[0077] Table 4 MTT test results of samples

[0078]

[0079] The Mean in Table 4 represents the mean relative cell viability of three replicate wells at each concentration gradient; the SD represents the standard deviation of the mean relative cell viability. According to the MTT results, the relative cell viability reached 90% or higher at concentrations below 5% (v / v) for the MGHP peptide sample group. Therefore, the MGHP peptide exhibited no significant cytotoxicity in macrophages within the 5% (v / v) concentration range.

[0080] Immuno-inflammatory tests:

[0081] 1) Cell seeding: After thawing cells, when the plating rate reaches approximately 60%, seed the cells into 6-well plates and incubate overnight in a CO2 incubator (37°C, 5% CO2).

[0082] 2) Solution preparation: Prepare the test substance working solution according to the test grouping (Table 5), where the sample group is the corresponding group in Table 4 with a volume concentration of 2.5% of the peptide MGHP.

[0083] Table 5 Test plan

[0084]

[0085] 3) Dosing: When the cell plating rate in the 6-well plate reaches 40%-60%, dosing will be performed according to the test group. Each group will have three replicate wells. For the blank control (NC) and negative control (BC), 1.8 mL of DMEM (Gibco) culture medium (1.8 mL) will be added to each well. For the positive control (PC), 1.8 mL of culture medium containing dexamethasone will be added to each well. For the sample group, 1.8 mL of culture medium containing the test sample (MGHP peptide and DMEM) at the corresponding concentrations listed in Table 5 will be added to each well. After dosing, the 6-well plate will be incubated in a CO2 incubator (37°C, 5% CO2) for 24 hours.

[0086] 4) LPS stimulation: 2 hours after administration, 200 μl of LPS-containing working solution was added to each well of the remaining groups except the blank control group, and the cells were placed in a CO2 incubator (37°C, 5% CO2) and cultured for 22 hours.

[0087] 5) Cell collection: After incubation, discard the old solution, wash twice with PBS, add 1 mL of RNAiso Plus to each well, pipette to lyse the cells, and collect the samples.

[0088] 6) ELISA test: Collect the cell culture supernatant and perform the test and analysis according to the operating instructions of the ELISA kit.

[0089] 7) Calculation of inhibition rate:

[0090] .

[0091] 8) Statistical Analysis: Results are expressed as mean ± SD. Comparisons between groups were performed using the t-test. All statistical analyses were two-tailed. P < 0.05 was considered a significant difference, and P < 0.01 was considered a highly significant difference. Test results are shown in Tables 6-8.

[0092] Table 6 Summary of IL-1β content detection results

[0093]

[0094] Note: When using the t-test method for statistical analysis, compared with the BC group, the significance is indicated by #, P-value < 0.05 is indicated by #, and P-value < 0.01 is indicated by ##. Compared with the NC group, the significance is indicated by *, P-value < 0.05 is indicated by *, and P-value < 0.01 is indicated by **.

[0095] Table 6 shows that compared to the BC group, the IL-1β level in the NC group increased significantly, indicating that the stimulation conditions in this test were effective. Compared to the NC group, the IL-1β level in the PC group decreased significantly, indicating that the positive control in this test was effective. Compared to the NC group, the IL-1β level in the sample peptide MGHP-2.5% decreased significantly, with an inhibition rate of 82.31%.

[0096] Table 7 Summary of TNF-α content test results

[0097]

[0098] As shown in Table 7, the TNF-α level in the NC group increased significantly compared to the BC group, indicating that the stimulation conditions in this test were effective. The TNF-α level in the PC group decreased significantly compared to the NC group, indicating that the positive control in this test was effective. The TNF-α level in the sample peptide MGHP-2.5% decreased significantly compared to the NC group, with an inhibition rate of 31.75%.

[0099] Table 8 Summary of PGE2 content test results

[0100]

[0101] Table 8 shows that the PGE2 content in the NC group significantly increased compared to the BC group, indicating that the stimulation conditions in this test were effective. The PGE2 content in the PC group significantly decreased compared to the NC group, indicating that the positive control in this test was effective. The PGE2 content in the sample peptide MGHP-2.5% was significantly decreased compared to the NC group, with an inhibition rate of 87.17%.

[0102] In summary, after LPS stimulation, compared with the negative control group, the content of inflammatory factors (IL-1β, TNF-a) and inflammatory mediators (PGE2) in the sample polypeptide MGHP at a concentration of 2.5% (v / v) was significantly decreased, with inhibition rates of 82.31%, 31.75%, and 87.17%, respectively. This shows that the sample at this concentration can inhibit the secretion of macrophage inflammatory factors (IL-1B, TNF-a) and inflammatory mediators (PGE2), thereby having the effect of alleviating the inflammatory response.

[0103] Furthermore, although exemplary embodiments have been described herein, the scope includes any and all embodiments based on the present application with equivalent elements, modifications, omissions, combinations (e.g., solutions that intersect various embodiments), adaptations, or changes. The elements of the claims are to be interpreted broadly based on the language employed in the claims and are not limited to the examples described in this specification or during the prosecution of the application, which examples are to be construed as non-exclusive. Therefore, it is intended that this specification and examples be considered merely as examples, with the true scope and spirit being indicated by the following claims and their full scope of equivalents.

[0104] The above description is intended to be illustrative and not restrictive. For example, the above examples (or one or more of their embodiments) may be used in combination with each other. For example, a person of ordinary skill in the art may use other embodiments when reading the above description. In addition, in the above detailed description, various features may be grouped together to simplify the application. This should not be interpreted as an intention that a feature of an application that is not claimed for protection is essential to any claim. On the contrary, the subject matter of the present application may have less than all the features of an embodiment of a particular application. Therefore, the following claims are incorporated into the detailed description as examples or embodiments, with each claim independently serving as a separate embodiment, and it is contemplated that these embodiments may be combined with each other in various combinations or arrangements. The scope of the invention should be determined with reference to the appended claims and the full scope of equivalents to which such claims are entitled.

[0105] The above embodiments are merely exemplary embodiments of the present application and are not intended to limit the present invention. The scope of protection of the present invention is defined by the claims. Those skilled in the art may make various modifications or equivalent substitutions to the present invention within the spirit and scope of protection of the present application, and such modifications or equivalent substitutions shall also be deemed to fall within the scope of protection of the present invention.

Claims

1. A use of a Spirulina phycocyanin polypeptide in the preparation of an anti-inflammatory product, characterized in that: The anti-inflammatory product is a cosmetic or a medicine. The Spirulina phycocyanin polypeptide sequence is shown in SEQ ID NO.

1. The anti-inflammatory effect is manifested in: inhibiting the secretion of macrophage inflammatory factors and inflammatory mediators.

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