CN113773366B

CN113773366B - Anti-inflammatory polypeptide BMP14, and preparation method and application thereof

Info

Publication number: CN113773366B
Application number: CN202110852520.5A
Authority: CN
Inventors: 何阳
Original assignee: Sichuan Liyan Workshop Biotechnology Co ltd
Current assignee: Sichuan Liyan Workshop Biotechnology Co ltd
Priority date: 2021-07-27
Filing date: 2021-07-27
Publication date: 2023-06-16
Anticipated expiration: 2041-07-27
Also published as: CN113773366A

Abstract

The invention provides an anti-inflammatory polypeptide BMP14, the amino acid sequence of which is Ile-Asn-Leu-Arg-Val-Ile-Ala-Cys-Leu-Val-Arg-Lys-Ile-Leu. It has good antibacterial and antiinflammatory effects and good stability. The invention also provides a preparation method of the anti-inflammatory polypeptide BMP14, which synthesizes crude polypeptide by using a solid phase synthesis method according to the sequence Ile-Asn-Leu-Arg-Val-Ile-Ala-Cys-Leu-Val-Arg-Lys-Ile-Leu. The preparation cost is low, the quality is high, and the prepared product has the above effects. The invention also provides application of the anti-inflammatory polypeptide BMP14 in the fields of cosmetics, medicines and skin care products.

Description

Anti-inflammatory polypeptide BMP14, and preparation method and application thereof

Technical Field

The invention relates to the technical field of polypeptides and application thereof, in particular to an anti-inflammatory polypeptide BMP14, and a preparation method and application thereof.

Background

Skin inflammation, mostly with pain, itching and redness; blisters, dryness, peeling, etc. may also be present due to severity. The common skin infection is staphylococcus aureus infection, which clinically manifests as skin furuncles, carbuncles, gangrene, cellulitis, folliculitis and the like.

Skin microorganisms are important members of the skin micro-ecosystem, and the flora on the skin surface can be generally divided into resident bacteria and transient bacteria. The resident bacteria are a group of microorganisms which colonize healthy skin, including staphylococcus, corynebacterium, propionibacterium, acinetobacter, malassezia, micrococcus, enterobacter, klebsiella, and the like. Skin resident bacteria are closely related to skin health. The resident bacteria can resist the growth of external pathogenic bacteria on the surface of the skin by regulating and controlling the expression of the antibacterial peptide by the skin keratinocytes.

Antibacterial peptides are an important component of the animal immune system. Under proper antibacterial concentration, the antibacterial peptide can interact with various microbial targets such as biological membrane/wall components or intracellular organelles, the former damages the integrity of cytoplasmic membranes, and the latter interferes with normal metabolic activities of cells, and finally bacterial death is caused. Under certain physiological conditions, the antibacterial peptide has activity related to inflammatory reaction, innate immunity and adaptive immunity in innate immune cells. The existing antibacterial peptide has biological activity, but has the defects of side effects such as hemolysis, cytotoxicity and the like, poor in-vivo stability and the like, and is difficult to realize clinical application.

Disclosure of Invention

The first aim of the invention is to provide an anti-inflammatory polypeptide BMP14, which has good antibacterial and anti-inflammatory effects and good stability;

the second object of the present invention is to provide a method for preparing anti-inflammatory polypeptide BMP14, which has the advantages described above;

it is a third object of the present invention to provide the use of the above anti-inflammatory polypeptide BMP14 in a variety of fields.

The invention is realized by the following technical scheme:

an anti-inflammatory polypeptide BMP14, the amino acid sequence of which is Ile-Asn-Leu-Arg-Val-Ile-Ala-Cys-Leu-Val-Arg-Lys-Ile-Leu.

The anti-inflammatory polypeptide BMP14 provided by the invention can act on cell membranes of bacteria, so that the cells are cracked, and an antibacterial effect is realized; can also influence the degranulation of mast cells to realize anti-inflammatory effect.

Specifically, BMP14 inhibits the conversion of arachidonic acid to prostaglandin E2 (PG E2) in leukocytes, achieving its anti-inflammatory effect. At high concentrations, BMP14 molecules occupy the hinge region of 2 IgE molecules cross-linked by antigen, resulting in a change in their configuration, causing a change in the normal configuration of the IgE-Fc receptor complex, thus inhibiting mast cell activity.

Further, the molecular weight is 1624.0Da and the isoelectric point is 10.86.

A preparation method of anti-inflammatory polypeptide BMP14 comprises synthesizing crude polypeptide according to sequence Ile-Asn-Leu-Arg-Val-Ile-Ala-Cys-Leu-Val-Arg-Lys-Ile-Leu by solid phase synthesis method.

Further, the method comprises desalting and purifying the crude polypeptide by HPLC reverse phase column chromatography, and identifying the purity until the purity of the polypeptide is not less than 95%.

Further, determining the molecular weight and isoelectric point of the purified polypeptide is also included.

Further, the method of HPLC purification is: the crude polypeptide was dissolved in ultrapure water containing 0.1% trifluoroacetic acid, mobile phase A was 0.1% trifluoroacetic acid-water, mobile phase B was 0.1% trifluoroacetic acid-acetonitrile, and after the baseline was stationary, loading was started, with a loading amount of 50. Mu.L.

The HPLC purified chromatographic column is a silica gel alkyl bonding phase C18 column, a binary mobile phase gradient elution system is adopted for gradient elution, namely, the content of mobile phase B in the eluent is increased from 0% -80% in a linear relation within 30min, the flow rate is 1mL/min, the detection wavelength 215nm is measured at 25 ℃.

Further, the molecular weight of the material is 1624.0Da by the determination of matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF), and the specific method is as follows: the purified polypeptide was dissolved in deionized water to prepare a 1. Mu. Mol/mL solution, and 10. Mu.L of the solution was mixed with an equal volume of saturated matrix solution (prepared by dissolving α -cyano-4-hydroxycinnamic acid in 50% acetonitrile containing 0.1% trifluoroacetic acid, and then subjecting the mixture to centrifugation, and collecting the supernatant).

Further, the isoelectric point of the purified polypeptide was determined to be 10.86 by isoelectric focusing electrophoresis, and the amino acid sequence structure of the purified polypeptide was determined by an automatic amino acid sequencer to be Ile-Asn-Leu-Arg-Val-Ile-Ala-Cys-Leu-Val-Arg-Lys-Ile-Leu.

The anti-inflammatory polypeptide BMP14 or the product prepared by the preparation method of the anti-inflammatory polypeptide BMP14 is applied to the field of cosmetic preparation.

Specifically, the anti-inflammatory polypeptide BMP14 is taken as a raw material, cosmetics are produced by adding cosmetic auxiliary materials and other raw materials, and common auxiliary materials are as follows: normal saline, pure water, mannitol, polyalcohol, glycerin, lanolin, mineral oil, vegetable oil, etc. Other cosmetic raw materials commonly used in this manner are combined with: other proteins, other polypeptides, plant extracts, cytokines, vitamins, etc. Specifically, one or more of auxiliary materials and cosmetic raw materials are selected for matching.

The anti-inflammatory polypeptide BMP14 or the preparation method of the anti-inflammatory polypeptide BMP14 is applied to the field of medicines for treating skin inflammation.

The anti-inflammatory polypeptide BMP14 or the preparation method of the anti-inflammatory polypeptide BMP14 is applied to the development field of skin microorganism regulation and antioxidant skin care products.

The technical scheme of the invention has at least the following advantages and beneficial effects:

(1) The invention provides an anti-inflammatory polypeptide BMP14, which has strong antibacterial, anti-inflammatory and antioxidant functions;

(2) The invention provides an anti-inflammatory polypeptide BMP14, which has good stability and low side effect;

(3) The invention provides a preparation method of anti-inflammatory polypeptide BMP14, which uses solid phase to synthesize polypeptide, has low cost and high quality controllability;

(4) The invention provides application of anti-inflammatory polypeptide BMP14 in the fields of cosmetics, medicines for treating skin inflammation and skin care product development.

Drawings

FIG. 1 shows the measurement of the hemolytic activity of BMP 14;

FIG. 2 shows the results of a cytotoxic activity assay of BMP 14;

FIG. 3 shows the results of plasma stability testing of BMP 14;

FIG. 4 is a test result of BMP14 to reduce the generation of ROS;

FIG. 5 shows the results of a test for the effect of BMP14 on inflammatory factors.

Detailed Description

Example 1

(1) According to the designed amino acid sequence:

Ile-Asn-Leu-Arg-Val-Ile-Ala-Cys-Leu-Val-Arg-Lys-Ile-Leu, and synthesizing to obtain crude polypeptide by solid phase synthesis;

(2) Desalting and purifying the crude polypeptide by HPLC reversed phase column chromatography, and identifying the purity until the purity of the polypeptide is not lower than 95%;

HPLC purification and identification method:

dissolving 0.1mg of a sample to be tested in 1mL of ultrapure water containing 0.1% trifluoroacetic acid, filtering with a 0.45 mu m filter membrane if undissolved impurities exist, wherein a mobile phase A is 0.1% trifluoroacetic acid-water, a mobile phase B is 0.1% trifluoroacetic acid-acetonitrile, and starting to sample after a base line is stable, wherein the sample loading amount is 50 mu L; the chromatographic column is a silica gel alkyl bonding phase C18 column (4.6mm×300mm, colloidal particle size 5 μm, pore size 100A), a binary mobile phase gradient elution system is adopted to perform gradient elution, namely the content of mobile phase B in the eluent is increased from 0% -80% in a linear relation within 30min, the flow rate is 1mL/min, the detection wavelength 215nm, and the detection temperature is 25 ℃.

(3) The molecular weight of the material is 1624.0Da by the determination of matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF);

the method comprises the following steps: the purified polypeptide was dissolved in deionized water to prepare a 1. Mu. Mol/mL solution, and 10. Mu.L of the solution was mixed with an equal volume of saturated matrix solution (prepared by dissolving α -cyano-4-hydroxycinnamic acid in 50% acetonitrile containing 0.1% trifluoroacetic acid, and then subjecting the mixture to centrifugation, and collecting the supernatant).

(4) The isoelectric point of the purified polypeptide is 10.86 by isoelectric focusing electrophoresis, and the amino acid sequence structure of the purified polypeptide is determined by an automatic amino acid sequencer, and is determined to be Ile-Asn-Leu-Arg-Val-Ile-Ala-Cys-Leu-Val-Arg-Lys-Ile-Leu.

Experimental example 1

Antibacterial Activity detection

The minimum inhibitory concentration is the lowest drug concentration capable of inhibiting bacterial growth and reproduction. The experiment adopts a method of double gradient dilution, and the specific experiment operation is as follows: fresh bacterial solutions were prepared, OD600 of the bacterial solutions was measured by an ultraviolet spectrophotometer, and the bacterial solutions were diluted to 2X105 CFU/ml with fresh LB liquid medium at 1OD600 = 1X 109 CFU/ml. Adding 100 mu l of physiological saline into a sterile 96-well plate in advance, adding a sample to be detected into a first well, sequentially carrying out double gradient dilution on the sample to be detected, adding 100 mu l of bacterial liquid with the concentration of 2X105 CFU/ml into each well, blowing and mixing uniformly by a pipetting gun, placing the mixed solution into a constant-temperature incubator at 37 ℃ for overnight culture, and finally detecting the light absorption value of the bacterial liquid at 600nm by using an enzyme-labeled instrument to detect the average value of the sample concentrations of the well and the adjacent wells where bacteria grow as the minimum inhibitory concentration, namely the MIC value. The bacteriostatic activity of the polypeptide BMP14 is shown in table 1.

Table 1: minimum inhibitory concentration of polypeptide BMP14 against standard strains

From Table 1, the minimum inhibitory concentrations of polypeptide BMP14 against the Staphylococcus aureus standard strain (Staphylococcus aureus ATCC 2592) and the Escherichia coli standard strain (Escherichia coli ATCC 25922) were 4.34. Mu.g/ml and 8.76. Mu.g/ml, respectively.

The experimental results show that the polypeptide BMP14 provided by the invention has good antibacterial effect.

Experimental example 2

Hemolytic activity and cytotoxicity assays

The density of the washed red blood cells is diluted and adjusted to 107-108/ml by using normal saline, meanwhile, samples to be tested are prepared into different gradient concentrations, the samples to be tested are placed at the constant temperature of 37 ℃ for co-incubation for 30min, then centrifugation is carried out at 1000rpm for 5min, and the light absorption value of the supernatant liquid at 540nm is detected by using an enzyme-labeling instrument. In this experiment, physiological saline was used as a negative control, the same volume of Triton X-100 (10%) was used as a positive control, and the hemolytic activity was proportional to the light absorption at 540 nm.

The hemolytic activity of BMP14 is lower as shown in figure 1.

Cytotoxicity assays were performed using the human embryonic kidney cell line HEK 293T. After the cells grew well and had a density of 80% of the bottom, the medium was discarded, washed 3 times with sterile PBS, then the adherent cells were digested with pancreatin, fresh 10% FBS-containing DMEM medium was added after termination and the cell suspension concentration was adjusted to 5X 105 cells/ml by pipetting, and 200. Mu.l of the cell suspension was added to each well using a sterile 96-well plate for the experiment, and the cells were cultured overnight in a cell incubator. The next day, samples to be tested were added at different concentrations, the concentration gradients were set at 80. Mu.g/ml, 40. Mu.g/ml, 20. Mu.g/ml, 10. Mu.g/ml, 5. Mu.g/ml, 0. Mu.g/ml, 3 replicates per concentration, the control group was sterilized PBS with the same volume, and then placed at 37℃with 5% CO ₂ The culture was continued for 24 hours in a constant temperature incubator. 10. Mu.l of MTT solution (5 mg/ml) was added to each well, and the mixture was placed in an incubator under dark conditions for 4 hours. Finally, the wells were carefully aspirated and discarded, DMSO (dimethyl sulfoxide) μl was added, the 96-well plates were placed on a shaker and slowly shaken for 10 minutes to dissolve the crystals, and the absorbance of each well at 490nm was measured using an microplate reader.

The test result is shown in figure 2, and the result shows that the anti-inflammatory polypeptide BMP14 provided by the invention has high cell proliferation capability and no cytotoxicity.

Experimental example 3

Stability test

10mL of human blood is obtained by a sterile syringe and stored in an anticoagulant tube, and centrifuged at 3500rpm at 4℃for 30min, and the yellow supernatant is carefully aspirated to obtain the desired plasma. The above plasma was diluted twice with sterile physiological saline, and the polypeptide BMP14 was added thereto, controlling the final concentration of BMP14 to 10mg/ml. Subsequently, the plasma in which the polypeptide BMP14 was dissolved was placed in a constant temperature incubator at 37℃for incubation, 10. Mu.l each was taken at 8 time points of 0, 0.5, 1, 2,4, 6, 8, 10 hours, etc., and the antibacterial activity against Acinetobacter baumannii was measured by the zone of inhibition method after co-incubation of the polypeptide BMP14 with the plasma, and two replicates were set for each time point.

As shown in the figure 3, the test result shows that the BMP is incubated with serum for 10 hours, and the antibacterial activity can still be detected, so that the stability of the BMP14 provided by the invention is good.

Experimental example 4

Influence on ROS

RAW264.7 cells (2X 105 cells/ml) with good log phase were plated on 96-well plates and cultured overnight in 40ml/L DMEM maintenance medium per well. The supernatant was discarded, the blank group was added with maintenance medium, the drug-treated group was first added with BMP14 maintenance medium containing 80, 40, 20ug/ml and the cells were incubated for 1h, then with maintenance medium containing LPS at a final concentration of 1ug/ml, and after 48h of incubation, the supernatant was discarded. Adding the ROS detection probe DCFH-DA into diluted cells, incubating for 15min at 37 ℃ with the final concentration of 10uM, washing the cells 400xg, and removing redundant probes; flow cytometry detects ROS-positive cell fractions using an excitation wavelength of 488nm and an emission wavelength of 525 nm.

As can be seen from fig. 4, LPS-induced ROS production can be significantly inhibited with increasing doses of BMP 14. The inhibition rate of ROS-positive (DCF-positive) cells was 23% in the 200ug/ml group, 39% in the 400ug/ml group, and 62% in the 800ug/ml group.

Experimental example 5

Effects on inflammation

The effect of BMP14 on cytokines was tested according to the instructions of the different cytokine ELISA kits. The ELISA kit is used for respectively detecting the contents of TNF-alpha, IL-6, IL-10, IL-1 beta and IFN-gamma in the culture medium supernatant. The specific process is as follows: the coated antibodies were diluted to a concentration of 0.5 and 2.0. Mu.g/ml with the coating solution, and 96-well ELISA plates, 100. Mu.l/well, 3 wells were repeated and coated overnight at 4 ℃. After the next day removal, the washing was performed three times with PBST for 5 minutes each. After blocking with 200. Mu.l of blocking solution 2% BSA at 37℃for 1h, the same washing as before was carried out. Cell supernatants stored at-20℃for detection were added to ELISA plates, 50. Mu.l/well, 48r and 72h culture supernatants were coated on wells, respectively, 50. Mu.l of wash solution was added to each well, placed in a wet box at 37℃for 2h, and removed and washed as before. Then 100. Mu.l of biotin-conjugated antibody diluted to the working concentration (1. Mu.g/ml) according to instructions was added to each well, and the mixture was placed in a wet box at 37℃for 2 hours, and the mixture was washed as before. Then the horseradish peroxidase of the avidin in each well100 μl of the chemoattractant enzyme-labeled secondary antibody (1:1000 dilution) was allowed to act in a37℃wet box for 1h, followed by washing as before. Then 100. Mu.L of a chromogenic solution (OPD) was added to each well and the mixture was allowed to act at 37℃for 15min, followed by the final addition of 50. Mu.L of a stop solution (2 MH ₂ SO ₄ ) The reaction was terminated. OD values were measured on a microplate reader.

FIG. 5 shows that BMP14 has significant inhibitory effects on secretion of LPS-stimulated TNF- α, IL-6, IL-10, and IL-1β; but BMP14 does not stimulate the production of TNF- α, IL-6, IL-10, IL-1 β, IFN- γ by the cells. The concentration of about 500. Mu.g/ml can inhibit the secretion of most cytokines, which indicates that the BMP14 of the present invention has good inflammation inhibiting effect.

Experimental example 6

Free radical scavenging Activity of BMP14 (DPPH radical scavenging assay)

DPPH (2, 2-diphenyl-1-picrylhydrazine) (2, 2' -diphenyl picrylhydrazine) is a stable nitrogen-centered lipid radical having 3 benzene rings in its structure, which is a purple solution of methanol or ethanol, has strong absorption around 517nm, and changes color from dark purple to pale yellow and has reduced absorbance at 517nm when an antioxidant with hydrogen donating ability is present. The degree to which the absorbance decreases has a positive correlation with the ability to scavenge free radicals and can therefore be used to detect antioxidant activity. A certain amount of DPPH (Sigma, USA) was weighed and dissolved in methanol to prepare a solution of 6X 10-5M for use. 48 μl of DPPH solution and 2 μl of sample (2 mg/ml) were mixed (mass ratio of final sample to DPPH is 3:1), left at room temperature for 30min at room temperature under dark conditions, and absorbance at 517nm was measured. The blank group replaces the sample to be tested with the sample dissolution medium. Three experiments were run in parallel and methanol was used when the uv spectrophotometer was zeroed. DPPH clearance (%) = (AB-AA)/ab×100 (AB: absorplanetan; AA: absorplanetan sample). At a sample concentration of 60. Mu.g/ml, the clearance of BMP-14 to DPPH free radicals was 48.2.+ -. 4.6%, indicating that BMP-14 had better free radical scavenging ability.

Experimental example 6

Effect of BMP-14 on mast cell degranulation

Wistar rats were sacrificed by cervical amputation, 10ml of Wash's solution (Tyrode's solution,137mM NaCl,2.7mM KCl,1.36mM NaCl 2,0.49mM MgCl2,0.36mM NaH2PO4,11.9mM NaH2CO3, and5.04mM D-glucose) was intraperitoneally injected, gently massaged for 10min, the abdominal cavity was opened to aspirate the Wash (containing mast cells), centrifuged at 1000rpm for 5min, the wash with Wash was repeated, and the mast cells were suspended with 1-2ml of Wash. 10 μl of the sample and 90 μl of the cell suspension were incubated at 37deg.C for 15min, and then centrifuged at 3000rpm for 5min. Mu.l of the supernatant was aspirated, 50. Mu.l of substrate (3 mg of rho-nitrophenyl-N-acetyl-. Beta. -D-glucosamidine in 10ml of 200mM, pH4.5 sodium acetate buffer) was added, incubation was continued for 6h, and the reaction was stopped by adding 100. Mu.l of Na2CO3 at pH10.0, and light absorption was measured at 405 nm. The negative control was sample dissolution medium and the positive control was Triton X-100 at 0.1% (v/v). This test was repeated 3 times. The mast cell degranulation rate was calculated according to the following formula: mast cell degranulation% = [ (amount of mast cell degranulation in sample group-amount of mast cell degranulation in negative control group)/amount of mast cell degranulation in positive control group ] ×100%. The degranulation activity of the mast cells of cathelicidin-BF revealed that BMP-14 had a degranulation rate of 42.5% at a concentration of 50. Mu.g/ml. The strong mast cell degranulation inhibition activity of BMP-14 suggests that it can affect immune responses and can play a role in inflammatory responses.

Experimental example 8

Effect of BMP14 on skin inflammation in rats

Carrying out dehairing on the backs of 40 SD rats 1 day before animal molding by using dehairing paste, wherein the area is 3cm multiplied by 3cm, and coating 50L 5%2, 4-Dinitrochlorobenzene (DNCB) on the dehairing area 1 day after dehairing for 1 st sensitization experiment; after 8 days, the dehairing area was dehaired twice, day 2, and 100L1% DNCB was applied outside the dehairing area for 2 nd sensitization. 3 weeks after, 100L1% DNCB was sensitized 1 time per week for 4 weeks. 3 days after the last sensitization, the molding is finished, and the skin surface of the rat shows symptoms such as keratinization, flaky erythema, crusting, prolongation of epidermis process, thickening of the acantha layer, slight spongy edema and the like, which indicates that the molding is successful. SD rats 30 successfully molded were selected and randomly divided into a model group (physiological saline), a positive control group (0.1% hydrocortisone butyrate cream, dose: 1.0 g/kg) and an experimental group (dose: 80 g/ear) according to a digital random table method, and 10 rats each were transdermally administered for 5 weeks. Acute inflammation, namely, ear-coating medicine on SD rats, continuously taking medicine for 5 weeks, and after taking medicine for 5 weeks, coating 20L 5% DNCB on the right ear for acute sensitization experiment. Efficacy assessment symptom score: rats were analyzed for skin lichenification, erythema and papule levels, and the severity of the symptomatic grading was scored. Mossiness, no mossiness at 0, small amount of fine scales at 1, obvious fine scales at 2, and large amount of scales at 3. The red spots are 0 minutes without red spots, and are invisible to about a little red spots, 2 minutes with obvious pale red spots and 3 minutes with a large amount of dark red spots. Pimple 0 is no pimple, 1 pimple is scattered, 2 pimples are fused and dense, and 3 pimples are fused obviously and dense. Prior to treatment, the three groups of rats had insignificant differences in the integral of symptoms of lichenification, erythema and papule (P > 0.05); after treatment, the positive control, experimental, and control groups were all significantly lower in the signs of lichenification, erythema, and papule scores than the model group (P < 0.05), as shown in Table 2.

TABLE 2 therapeutic effect of BMP14 on rat skin inflammation model

P < 0.05, the difference is obvious compared with the model group after treatment; the difference between the treatment and the pre-treatment is significant, and the difference between the treatment is less than 0.05.

The result shows that the brand new polypeptide BMP14 designed by the invention can directly kill pathogenic bacteria, and can regulate and control inflammatory factors by reducing ROS, influencing mast cell degranulation and the like, thereby reducing inflammatory reaction. Has obvious functions and rich action ways. The BMP14 has low hemolytic activity, no cytotoxicity and high stability, and can be applied to medicines or skin care products for treating skin inflammation.

The above is only a preferred embodiment of the present invention, and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

<110> Sichuan beauty Co., ltd

<120> an anti-inflammatory polypeptide BMP14 and method for preparing same

<160> 1

<170> PatentIn version 3.3

<210> 1

<211> 14

<212> PRT

<213> artificial sequence

<400> 1

Ile Asn Leu Arg Val Ile Ala Cys Leu Val Arg Lys Ile Leu

1 5 10

Claims

1. An anti-inflammatory polypeptide BMP14, characterized in that its amino acid sequence is lie-Asn-Leu-Arg-Val-lie-Ala-Cys-Leu-Val-Arg-Lys-lie-Leu.

2. The anti-inflammatory polypeptide BMP14 of claim 1 wherein the molecular weight is 1624.0Da and the isoelectric point is 10.86.

3. A preparation method of anti-inflammatory polypeptide BMP14 is characterized in that a solid-phase synthesis method is used for synthesizing crude polypeptide according to the sequence Ile-Asn-Leu-Arg-Val-Ile-Ala-Cys-Leu-Val-Arg-Lys-Ile-Leu.

4. The method of producing the anti-inflammatory polypeptide BMP14 according to claim 3, further comprising desalting and purifying the crude polypeptide by HPLC reverse phase column chromatography.

5. The method of claim 4, further comprising determining the molecular weight and isoelectric point of the purified polypeptide.

6. Use of the anti-inflammatory polypeptide BMP14 according to any one of claims 1-2 or the product prepared by the method for preparing the anti-inflammatory polypeptide BMP14 according to any one of claims 3-5 in the field of cosmetic preparation.

7. Application of the anti-inflammatory polypeptide BMP14 according to any one of claims 1-2 or the product prepared by the preparation method of the anti-inflammatory polypeptide BMP14 according to any one of claims 3-5 in the preparation of medicines for treating skin inflammation.

8. Use of an anti-inflammatory polypeptide BMP14 according to any one of claims 1-2 or a product prepared by a method for preparing an anti-inflammatory polypeptide BMP14 according to any one of claims 3-5 for preparing a product for regulating skin microorganisms, an antioxidant skin care product.