CN113248573A

CN113248573A - Application of active peptide and mesenchymal stem cell exosome for improving skin physiological characteristics in medicines or cosmetics

Info

Publication number: CN113248573A
Application number: CN202110536968.6A
Authority: CN
Inventors: 王京; 张然
Original assignee: Beijing Daiyu Biotechnology Co ltd
Current assignee: SHENZHEN HOPELIFE BIOTECHNOLOGY CO.,LTD.
Priority date: 2021-05-18
Filing date: 2021-05-18
Publication date: 2021-08-13
Anticipated expiration: 2041-05-18
Also published as: CN113248573B

Abstract

The invention belongs to the technical field of biological pharmacy, and particularly relates to application of active peptide for improving skin physiological characteristics and a mesenchymal stem cell exosome in medicines or cosmetics.

Description

Application of active peptide and mesenchymal stem cell exosome for improving skin physiological characteristics in medicines or cosmetics

Technical Field

The invention belongs to the technical field of biology, and particularly relates to application of active peptide and mesenchymal stem cell exosome for improving skin physiological characteristics in medicines or cosmetics.

Background

Skin aging, which is an important subject in the research of skin health care and skin care cosmetics at present, not only seriously affects the beauty but also may have specific symptoms such as dryness, roughness, telangiectasia, elasticity reduction, irregular pigmentation, seborrheic keratosis and the like. According to the action mechanism, skin aging is mainly divided into endogenous aging and exogenous aging, wherein the endogenous aging is a physiological process influenced by genes, hormones and the like, the exogenous aging mainly refers to the aging of the skin caused by exogenous physical and chemical stimulation, and ultraviolet rays are important factors for causing the skin aging.

The ultraviolet rays can be divided into 3 wave bands according to the wavelength, namely long-wave ultraviolet UVA (320-400nm), medium-wave ultraviolet UVB (275-320nm) and short-wave ultraviolet UVC (230-275nm), the UVC is almost completely absorbed by the ozone layer due to the blocking effect of the atmospheric layer, the ultraviolet rays incident to the earth surface are mainly UVA and UVB, and researches show that the damage of the same dosage of UVB radiation to the skin is about 800-1000 times larger than that of the UVA, and the damage to the skin is mainly UVB. When the skin is irradiated by excessive ultraviolet rays, oxidative stress is generated in skin tissue cells, excessive Reactive Oxygen Species (ROS) are generated, the ROS can destroy the integrity of cell membranes and an antioxidant system in cells, so that cell inflammation, cell apoptosis and tumor formation are caused, meanwhile, the excessive ultraviolet rays can degrade skin collagen, hyaluronic acid and elastin, so that the skin becomes low in elasticity, rough and wrinkles are formed, and if the skin is irradiated by ultraviolet rays for a long time, the skin can be excessively settled, and serious people can induce skin canceration.

Antioxidant stress is an important way for protecting skin from being damaged by ultraviolet rays, a plurality of antioxidant measures are provided for researchers, and the antioxidant measures are used for protecting skin and promoting repair after damage, for example, researchers in China currently propose that natural plant components such as raspberry (raspberry) extract, aloe extract, ginsenoside, neferine and the like are used for resisting oxidative stress caused by ultraviolet rays, and the researchers also propose that active peptides are used for protecting skin from ultraviolet damage, for example, Chinese patents CN202011275745, CN201810524802, CN201711441206 and CN201680062991 and the like all provide technical schemes for using the active peptides to protect ultraviolet damage, but the ultraviolet protection effect of the active peptides still needs to be enhanced, particularly, the efficacy of resisting oxidative stress is improved, the repair process after damage is promoted, the collagen secretion of the skin is improved, and the elasticity and the luster of the skin are rapidly recovered.

Stem cell therapy is a new treatment means which develops rapidly in recent years, and stem cells have strong multidirectional differentiation capacity, so that the stem cells can be used for treating various diseases, particularly in the field of skin care products, stem cells and derivative products thereof, such as stem cell culture solution, stem cell lysate, stem cell exosomes and the like are widely applied, for example, Chinese patent CN202011445563 requires protection of a composition containing umbilical cord mesenchymal stem cell exosomes and application thereof, and on the basis of adding the umbilical cord mesenchymal stem cell exosomes, the stem cell therapy can accelerate metabolism of skin cells, recover antioxidant and regeneration capacity of basal cells, prevent skin aging, increase skin elasticity and plumpness, reduce pores and improve gloss by matching with a humectant, an antioxidant and algal polysaccharide; chinese patent CN202010647884 claims a cosmetic composition and cosmetics, whose main ingredient is CD200 modified mesenchymal stem cell exosome, which can increase skin cell activity, promote skin cell metabolism, promote skin repair, and delay skin aging after being applied on skin surface; chinese patent CN201910819093 claims a method for simply preparing a cytokine from placental mesenchymal stem cells, optimizes a preparation method of a cytokine derived from placental mesenchymal stem cells, can prepare the cytokine on a large scale, and promotes repair and skin health care after skin injury. However, the components of the stem cell product are complex, the preparation process needs to be strictly controlled, and the stability of the skin care effect of the single use of the stem cell or the derivatives thereof is not good enough, so that the application of the stem cell in skin care cosmetics is limited to a certain extent.

Based on the problems in the prior art, the invention provides an active peptide with a brand-new amino acid structure, which can promote the proliferation of skin epithelial cells and fibroblasts, improve the oxidation resistance of cells, resist oxidative damage caused by external factors such as ultraviolet rays and the like, promote the repair of skin tissues, induce the formation of collagen substances and restore the normal physiological function of skin; the active peptide and the mesenchymal stem cell exosome can achieve a surprising synergistic effect, so that the capacities of resisting oxidative stress and promoting skin tissue repair are further enhanced.

Disclosure of Invention

The invention mainly aims to provide a new means capable of resisting skin damage caused by ultraviolet rays, in particular to an active peptide with a brand-new structure, and the peptide and mesenchymal stem cell exosome are combined and applied to cosmetic products, so that oxidative damage can be resisted, skin tissue repair is promoted, and the effects of maintaining beauty and keeping young are achieved.

The detailed technical scheme of the invention is as follows:

an active peptide capable of improving skin physiological characteristics is provided, and the amino acid sequence of the active peptide is shown as SEQ ID NO: 1 is shown.

A composition capable of improving the physiological properties of skin is provided, which is characterized by comprising the active peptide and mesenchymal stem cell exosomes.

Further, the mass ratio of the active peptide to the mesenchymal stem cell exosome is 1:10-10: 1.

Further, the mesenchymal stem cells are derived from fat, bone marrow, umbilical cord or placenta.

Further, the mesenchymal stem cells are derived from umbilical cord.

Further, the preparation method of the mesenchymal stem cell exosome comprises the following steps: separating mesenchymal stem cells from umbilical cord tissues, carrying out subculture, culturing for 3-5 generations until the fusion degree reaches more than 80%, collecting supernatant, centrifuging, filtering, and collecting.

In the prior art, a plurality of technical schemes for applying mesenchymal stem cell exosomes to skin health care or beauty treatment are reported, but most of the technical schemes use the stem cell exosomes alone, the exosomes are micro vesicles coated by biological membranes, and the exosomes contain nucleic acid, protein, polypeptide, saccharide and various active substances such as other stem cell metabolites and are rich in nutrition and diverse in physiological activity, but the components and the action mechanism are complex, and the expected protection effect is often difficult to achieve by using the stem cell exosomes alone. Therefore, the bioactive peptide and the mesenchymal stem cell exosome are used in a combined manner, so that the antioxidant stress effect of the bioactive peptide and the mesenchymal stem cell exosome can be effectively exerted, the characteristics of rich nutrition and easy absorption and utilization in the mesenchymal stem cell exosome can be utilized, the repair process after the damaged tissue is promoted, and the skin cells or the tissue can quickly recover the physiological function.

Provides an application of active peptide or related composition in preparing cosmetics.

Further, the cosmetic is an ointment, cream or spray.

The active peptide or the composition can be used for preparing various cosmetics, including but not limited to ointment, cream or spray, and specifically can be one or more of facial masks, eye cream, facial cream, essence, facial cleanser, stock solution, toner, emulsion, pure dew, sun block, BB cream, CC cream and sun block.

The beneficial effects of the invention include:

the invention provides a novel active peptide, which can promote the proliferation of skin epithelial cells and fibroblasts, effectively resist skin damage caused by oxidative stress, improve the oxidation resistance in a body, and promote the repair of skin tissues and the quick recovery of normal physiological functions; the composition is combined with mesenchymal stem cell exosomes, can further strengthen the antioxidant effect, and particularly can effectively promote skin cell proliferation, promote metabolism, delay the skin aging process and protect skin tissues from various injuries.

Drawings

FIG. 1 HaCaT cell viability plots;

FIG. 2 is a graph of the level of SOD change in a model of ultraviolet damaged HaCaT cells;

FIG. 3 is a graph of CAT levels in a model of UV-damaged HaCaT cells;

FIG. 4 is a graph of the level of MDA changes in the model of UV-damaged HaCaT cells;

FIG. 5 is a graph of HA change levels in the UV-damaged HFF-1 cell model;

FIG. 6 pathological sections of different treatment groups in the model of the skin of the ultraviolet injured rat;

FIG. 7 is a graph of SOD change levels in a model of UV-damaged rat skin;

FIG. 8 is a graph of MMP-1 levels in a model of UV-damaged rat skin.

Detailed Description

The following non-limiting examples are presented to enable those of ordinary skill in the art to more fully understand the present invention and are not intended to limit the invention in any way. All the technologies implemented based on the above-mentioned contents of the present invention should fall within the scope of the claims of the present application.

The experimental methods described in the following examples are all conventional methods unless otherwise specified; the reagent biomaterials, test kits, if not specifically indicated, are commercially available.

Example 1 preparation of active peptides for improving physiological properties of skin

The invention combines the earlier stage work foundation and bioinformatics resources, designs the molecular structure of a novel active peptide on the basis of preliminary experiments, the amino acid sequence of the active peptide is shown as SEQ ID NO.1 and is named as SDH peptide, and the active peptide is prepared by adopting a solid phase synthesis method, and the specific steps are as follows:

(1) taking Fmoc-Leu-Wang resin as a raw material, weighing 500mg of Fmoc-Leu-Wang resin, placing the Fmoc-Leu-Wang resin in a solid phase synthesis reaction tube, firstly washing the Fmoc-Leu-Wang resin with DMF for 2 times, then adding 15mL of DMF, soaking and swelling the mixture for 15-30min, washing the mixture for 3 times with DMF, and draining the mixture for later use.

(2) Adding 5mL DMF solution containing 150mg (0.5mmol) Fmoc-amino acid, 100mg (0.5mmol) HOBt, 200mg (0.5mmol) DMAP and 100 μ L (0.8mmol) DIC into the resin, placing in a constant temperature oscillator, reacting at room temperature for 3-4h, draining the reaction solution after reaction, washing the resin with DMF for 3 times each, taking a small amount of resin, and detecting with ninhydrin to judge (if the resin is colorless and transparent, the reaction is complete); then 5mL of 20% piperidine/DMF solution is added for reaction for 10min, the Fmoc protection of the N end on the alanine is removed, the resin is washed 3 times by dichloromethane and DMF, a small amount of resin is taken, and the color development is detected by ninhydrin solution.

(3) Repeating the above condensation method, dissolving Fmoc-amino acid (0.5mmol), HOBt (0.5mmol) and DIC (100 μ L) in 5ml NMP according to the predetermined amino acid sequence, sequentially coupling, detecting the reaction progress with ninhydrin reagent, and if the condensation is complete, the resin is colorless or pale yellow; in the case of the purple color, the condensation time is prolonged or the condensation is repeated. After the synthesis of the peptide chain is finished, washing the peptide chain with absolute ethyl alcohol for 8 to 10 times, and then blowing the resin with nitrogen to dry to obtain a crude polypeptide product with a specific amino acid sequence.

(4) 30g of crude polypeptide product is weighed, water is added for stirring, the pH value is adjusted to 8.0 by ammonia water until the crude polypeptide product is completely dissolved, and the solution is filtered by a 0.45 mu m mixed microporous filter membrane for later use. Purifying by adopting a high performance liquid chromatography, wherein a mobile phase system is 0.1 percent TFA/water solution-0.1 percent TFA/acetonitrile solution, the flow rate of a chromatographic column of 77mm multiplied by 250mm is 90ml/min, eluting by adopting a gradient system, circularly injecting and purifying, sampling a crude product solution in the chromatographic column, starting mobile phase elution, collecting a main peak, and evaporating acetonitrile to obtain an SDH peptide purified product.

(5) The purity of the polypeptide is up to more than 98% by HPLC-mass spectrum identification.

Example 2 preparation of umbilical cord mesenchymal stem cell exosomes

2.1 isolation and culture of human umbilical cord mesenchymal Stem cells (hMSCs)

Fresh umbilical cords were collected under sterile conditions and placed in sterile PBS solution containing antibiotics and maintained at 4 ℃. Under a sterile environment, the umbilical cord tissue is taken out, and is repeatedly washed by adopting a sterile PBS solution to remove residual blood and other impurities. Using sterile surgical scissors, cut the umbilical cord to about 1mm³Tissue pieces of size, the resulting tissue pieces were filtered through a 200 mesh screen to remove smaller tissue debris and cellular debris. Preparing DMEM/F12 culture medium, filtering for sterilization, adding 10% fetal calf serum, uniformly spreading the treated umbilical cord tissue block in a culture dish containing the culture medium, and placing the culture dish at 37 deg.C and 5% CO₂Culturing in an incubator.

Observing the growth condition of the cells by using an inverted microscope, and slightly rinsing the cells by using a serum-free culture medium according to the condition of a better culture medium, so as to remove tissue residues and cell fragments. And when the adherent cells in the culture dish are more than 60% full, digesting with 1mL of 0.25% recombinant trypsin for 3-5 minutes, adding 5-10mL of serum-containing culture medium to stop digestion, slightly blowing and beating into single cell suspension by using a pipette, centrifuging for 5 minutes at 1000rpm, removing supernatant, and re-suspending with 5-10mL of fresh culture medium to obtain the primary umbilical cord mesenchymal stem cells. And (3) subculturing the primary mesenchymal stem cells, and performing subsequent operation after culturing for 3-5 generations.

2.2 obtaining exosomes from hMSCs

Inoculating the subcultured hMSCs into DMEM/F12 medium containing 10% fetal bovine serum at 37 deg.C and 5% CO₂Culturing until the cell fusion degree reaches above 80%, sucking out the culture medium, washing with sterile PBS for 3 times, and replacing serum-free mediumCulturing in culture medium for 24 hr, and collecting supernatant; centrifuging the obtained supernatant at 2000rpm for 30min to remove cell debris, collecting supernatant, filtering with 0.22 μm sterile filter membrane to obtain filtrate, centrifuging at 10000rpm for 45min, discarding supernatant, and resuspending the obtained precipitate, i.e. human umbilical cord MSCs exosomes, with appropriate amount of PBS; the obtained exosome is subjected to NTA, transmission electron microscope observation and flow cytometry detection, and the result shows that the separated exosome can positively express CD90, CD63 and CD73 and negatively express CD34 and CD45, has a vesicular structure, has an average diameter of about 100-120nm, and meets the requirements of subsequent experiments.

Example 3 SDH peptides and mesenchymal Stem cell exosomes enhance epithelial cells' ability to fight oxidative stress

3.1 preparation of ultraviolet-damaged HaCaT cell model

DMEM medium containing 10% FBS at 37 deg.C and 5% CO₂HaCaT cells were cultured in an incubator. Taking HaCaT cells in a good state in an exponential growth phase, digesting, counting, inoculating the cells into a 96-well plate, wherein the inoculation concentration is 1 multiplied by 10⁵One/well, placed at 37 ℃ in 5% CO₂The cultivation was continued in the incubator for 12 hours. Irradiating the HaCaT cells with UVB (311nm) at an intensity of 50mJ/cm²And establishing an ultraviolet ray damage model.

3.2 cell survival assay

After the preparation of the uv damage model was completed, the cells were divided into 5 groups, which were: blank control, HaCaT cells without UVB irradiation, and the remaining 4 groups were irradiated cells, i.e.: PBS group, 100 μ L PBS was added; SDH group, adding 100 μ L10 mg/mL SDH polypeptide into each hole; adding 100 mu L of 10mg/mL hMSCs exosomes into each hole of the hMSCs group; combination group: adding 100 μ L of mixed solution of SDH polypeptide and hMSCs exosome into each well (wherein the concentration of SDH polypeptide and hMSCs exosome is 10mg/mL), placing at 37 deg.C, and 5% CO₂The cultivation was continued in the incubator for 24 hours.

The cell viability was determined by the CCK-8 method, according to the instructions of the CCK-8 test kit, 10. mu.L of CCK-8 solution was added to each well in 5% CO₂And incubating for 2h at 37 ℃, and measuring a light absorption value by using an enzyme-labeling instrument at a wavelength of 450nm so as to calculate the survival rate of the cells. As shown in fig. 1, is receiving UAfter VB irradiation, the cell survival rate of the PBS group is greatly reduced, which indicates that HaCaT cells are seriously damaged; after the SDH polypeptide and the hMSCs exosomes are applied, the survival rate of the HaCaT cells is obviously restored, and the improvement efficiency of the SDH polypeptide is slightly stronger than that of the hMSCs exosomes; in the experimental group simultaneously applying DH polypeptide and hMSCs exosome, the survival rate of HaCaT cells is greatly increased, which shows that the HaCaT cells and the hMSCs exosome can generate synergistic effect to accelerate the repair process of the HaCaT cells after damage.

3.3 antioxidant Capacity testing

In order to detect the antioxidant capacity of the active peptide and the mesenchymal stem cell exosome provided by the invention, the supernatant of the HaCaT cell 24 hours after the treatment is collected (the administration mode is the same as that in section 3.2), and the levels of Superoxide dismutase (SOD), Catalase (CAT) and Malondialdehyde (MDA) in the supernatant are respectively detected according to the method described in the kit specification.

As shown in fig. 2, the SDH polypeptide and the hMSCs exosomes can significantly increase the SOD expression level of HaCaT cells after ultraviolet damage, and the SDH polypeptide has a stronger improving ability than the hMSCs exosomes, and this trend is further strengthened in the combination treatment group; CAT is another important metabolite against oxidative damage in cells or organisms, as shown in FIG. 3, the levels of CAT secretion in SDH polypeptide, hMSCs exosome and combination treatment group are all obviously improved, but no statistical difference occurs between the three, which indicates that the three have similar efficacy in improving CAT secretion; MDA is an important metabolite in the oxidative damage process, and the excessive accumulation of MDA can cause cell or tissue damage, as shown in figure 4, after SDH polypeptide and hMSCs exosome are applied, the MDA level in HaCaT cell culture solution is obviously reduced, the reduction range of a combined treatment group is the largest, and the expression levels of SDH polypeptide and hMSCs exosome are similar. According to the analysis, the SDH polypeptide and the hMSCs exosome can resist oxidative damage caused by ultraviolet rays from different layers, and the synergistic treatment of the SDH polypeptide and the hMSCs exosome is more advantageous.

Example 4 SDH peptides and mesenchymal Stem cell exosomes promote collagen secretion

In order to verify the influence of the active peptide and the mesenchymal stem cell exosome provided by the invention on skin collagen secretion, an ultraviolet injury model is constructed by taking skin fibroblast HFF-1 as a research object, and the hyaluronic acid secretion condition of the model is verified.

4.1 preparation of UV-damaged HFF-1 cell model

DMEM medium containing 10% FBS at 37 deg.C and 5% CO₂HFF-1 cells were cultured in an incubator. Collecting HFF-1 cells in exponential growth phase, digesting, counting, inoculating the cells into 96-well plate at a concentration of 1 × 10⁵One/well, placed at 37 ℃ in 5% CO₂The cultivation was continued in the incubator for 12 hours. Irradiating the HaCaT cells with UVB (311nm) at an intensity of 50mJ/cm²And establishing an ultraviolet ray damage model.

4.2 measurement of hyaluronic acid secretion ability

Cell supernatants 24 hours after the above treatments were collected and the Hyaluronic Acid (HA) level in the supernatants was measured according to the method described in the kit instructions. As shown in fig. 5, the secretion of hyaluronic acid by fibroblasts is severely reduced by ultraviolet radiation, which induces skin roughness, chapping and formation of wrinkles, and this trend is suppressed and reversed by applying SDH polypeptide and hMSCs exosomes provided in the present invention, the content of hyaluronic acid in cell supernatant is increased, and the improvement effect seems to be more significant in the combination treatment group.

Example 5 SDH peptide and mesenchymal Stem cell exosomes protect rat model skin lesions

5.1 preparation and treatment of rat ultraviolet injury model

Healthy SD rats with SPF level, half male and female, weight of 180 +/-20 g are selected, and the groups are randomly divided into 10 groups, and the groups total 5 groups comprise a blank control group, a normal saline group, an SDH group, an hMSCs group and a combined group. The back hair of each group of rats was shaved to an area of about 5cm × 5cm, the rats were placed on a fixed platform and irradiated under an ultraviolet lamp (wavelength 311nm) at an irradiation distance of about 10cm once a day for 90min each time for 12 weeks, with the blank control group not subjected to ultraviolet irradiation.

Before ultraviolet irradiation each time, therapeutic agents are respectively smeared on the unhairing part, and the method specifically comprises the following steps: the normal saline group is smeared with 500 mu L of normal saline; SDH group, smearing 20mg/kg SDH polypeptide; coating 20mg/kg of hMSCs exosomes on the hMSCs group; the combined group is coated with 20mg/kg SDH polypeptide and 20mg/kg hMSCs exosome and continuously treated for 12 weeks.

5.2 pathological examination of rat skin

After the rat model was continuously treated for 12 weeks, the rats were decapped and the skin tissue of the epilated portion of the back of the rat was taken to be about 1cm²Fixing in 4% multi-poly fermentation solution for 48 hr, embedding in paraffin, preparing pathological section, and HE staining. As shown in FIG. 6, the normal nude mouse has intact skin tissue structure and normal thickness, and has the advantages of regular arrangement of the horny layer, the granular layer, the acanthocyte layer and the basal layer, and obvious and clear layering of the cells. After ultraviolet irradiation, the epidermis layer is disordered, the cells of the dermis layer are reduced and arranged disorderly, the blood vessels are expanded obviously, partial hyperemia is caused, and the infiltration of lymphocytes and eosinophils is caused obviously. After SDH polypeptide and hMSCs exosome treatment, the skin injury symptom is obviously improved, particularly in a combined group, the layered structure of the skin is clear, fibroblasts in the dermis layer are increased, infiltration of lymphocytes and eosinophils is reduced, and the gland and hair follicle structure is gradually recovered.

5.3 antioxidant Capacity testing

After the rat model is continuously treated for 12 weeks, the rat is subjected to neck removal treatment, skin tissues of a depilated part on the back of the rat are taken and cleaned by sterile normal saline, 0.5g of tissues are taken and cut into pieces by using sterile surgical scissors, about 5mL of sterile PBS solution is added, a homogenizer is used for full grinding to obtain tissue homogenate, the tissue homogenate is centrifuged at 4 ℃ and 4000r/min for 10 minutes, and the supernatant is collected for subsequent detection.

The level of SOD in the supernatant was detected according to the method described in the kit specification, as shown in fig. 7, in rat skin tissue, after uv damage, the SOD content in the negative control group (saline group) was significantly reduced, only about half of that in normal tissue, and after SDH polypeptide and hMSCs exosome treatment, there was recovery in different degrees, where the antioxidant effect of hMSCs exosome was stronger than that of SDH polypeptide, unlike in cell experiments, probably because the physiological environment and regulatory factors of skin tissue are more complex, and hMSCs exosome is rich in various nutrients and regulatory substances, so it showed stronger antioxidant ability in skin tissue, and in combination treatment group, the expression level of SOD was higher, indicating that this combination in skin tissue also can obtain significant synergy, consistent with the effect of cell level experiments.

5.4 MMP-1 expression level detection

Matrix metalloproteinase 1 (MMP-1) is the most important enzyme for causing aging symptoms such as wrinkles and the like of skin, and is closely related to synthesis and secretion of other metalloproteases, an enzyme-linked immunosorbent assay (ELISA) kit is adopted in the invention to properly adjust the MMP-1 level in skin tissue homogenate, and the specific operation method is carried out according to the kit specification.

As shown in figure 8, after ultraviolet irradiation, the MMP-1 expression level in the skin is obviously increased and reaches more than 2 times of the normal level, although the MMP-1 expression level is reduced after the treatment of the SDH polypeptide and the hMSCs exosomes, when the reduction range is limited, the MMP-1 expression level is further reduced after the combined treatment of the two substances, which fully shows that the composition of the SDH polypeptide and the hMSCs exosomes can effectively inhibit the secretion and expression of MMP-1, inhibit skin aging and protect skin tissues from being damaged by the ultraviolet radiation.

Sequence listing

<110> Beijing Biotechnology Ltd

<120> application of active peptide for improving skin physiological characteristics and mesenchymal stem cell exosome in medicine or cosmetics

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<170> SIPOSequenceListing 1.0

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<213> Artificial Sequence (Artificial Sequence)

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Glu Val Lys Leu Gly Asn Lys Leu Ser Pro Ala Leu Gly Glu Pro Leu

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Cys Thr Ser His Lys

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Claims

1. An active peptide capable of improving the physiological properties of the skin, characterized by an amino acid sequence as set forth in SEQ ID NO: 1 is shown.

2. A composition capable of improving the physiological properties of skin, comprising the active peptide of claim 1 and mesenchymal stem cell exosomes.

3. The composition of claim 2, wherein the mass ratio of the active peptide to the mesenchymal stem cell exosomes is 1:10-10: 1.

4. The composition of claim 2, wherein the mesenchymal stem cells are derived from fat, bone marrow, umbilical cord, or placenta.

5. The composition of claim 4, wherein the mesenchymal stem cells are derived from umbilical cord.

6. The composition of claim 5, wherein the mesenchymal stem cell exosomes are prepared by a method comprising: separating mesenchymal stem cells from umbilical cord tissues, carrying out subculture, culturing for 3-5 generations until the fusion degree reaches more than 80%, collecting supernatant, centrifuging, filtering, and collecting.

7. Use of an active peptide as claimed in claim 1 or of a composition as claimed in claims 2 to 6 for the preparation of a cosmetic product.

8. Use according to claim 7, characterized in that the cosmetic product is an ointment, cream or spray.