Use of composition comprising cryopreserved prp in treatment of gynecological diseases

Abstract

The invention provides an application of a composition containing cryopreserved prp in treatment of gynecological diseases. The invention also provides a reproduction promoting peptide which can obviously promote the proliferation and migration activity of human endometrial stromal cells, can reduce the apoptosis level of the cells, and has a better promoting effect. In addition, the cryopreserved prp is also prepared, after the cryopreserved prp is combined with the reproduction promoting peptide, the repair of the endometrial injury of the rat can be remarkably promoted, and a relatively good treatment effect is achieved.

Description

Technical Field

[0001] The present application relates to the field of biopharmaceuticals, and specifically to the use of a composition containing cryopreserved prp in the treatment of gynecological diseases. Background Art

[0002] The endometrium is the soil that breeds life and is a key component of women's reproductive health. When the endometrium is severely damaged, such as under pathological conditions such as repeated curettage and infection, the basal layer of the uterus is damaged, and the proliferation capacity of the basal layer cells is impaired. The most direct manifestation is the thinning of the endometrium. At the same time, severely damaged endometrium may undergo pathological repair during the repair process, resulting in varying degrees of fibrosis, and in severe cases, even leading to uterine cavity adhesions and uterine cavity occlusion. Endometrial damage can easily lead to reduced menstruation, amenorrhea, repeated miscarriages and infertility, and may manifest as repeated implantation failure (RIF) during in vitro fertilization and embryo transfer. When only the functional layer of the endometrium is slightly damaged, the epithelial cells on the wound surface can quickly regenerate and repair the endometrium without scars. Severe damage to the basal layer of the endometrium will cause continuous deposition of collagen fibers. On the one hand, the collagen fiber deposition covers the surface of the basal layer, and the basal endometrial stem cells cannot proliferate and regenerate. On the other hand, the new scar tissue usually has no vascular structure, and insufficient blood supply further hinders the regeneration and repair of the endometrium. Therefore, it reduces fibrous scar formation and promotes the regeneration and repair of damaged endometrium to restore the normal physiological function of the uterine cavity structure and endometrium.

[0003] Drug therapy is the most commonly used treatment for endometrial damage and repair. Estrogen: Estrogen receptors exist in the endometrium. Estrogen can promote endometrial regeneration by inhibiting epithelial-mesenchymal transition (EMT) induced by transforming growth factor-β (TGF-β) and activating the Wnt / β-catenin signaling pathway. Therefore, estrogen is often used as an adjuvant therapy after some intrauterine operations to prevent intrauterine adhesions. However, there are currently no recognized standards for the dosage, timing, route of administration, and drug safety of estrogen for preventing uterine adhesions. Vasoactive drugs: Good blood flow is a necessary condition for endometrial growth and repair. Some vasoactive drugs such as sildenafil citrate can act on uterine artery smooth muscle, selectively inhibit phosphodiesterase 5 (PDE5), enhance the dilation effect of nitric oxide on vascular smooth muscle, increase uterine artery blood flow, and improve endometrial development, bringing hope of fertility to patients with severe IUA who have fertility requirements. However, there is currently a lack of a large amount of conclusive research evidence to support this, and these therapeutic effects are not included in the drug instructions. The use of vasoactive drugs is somewhat controversial and still requires further in-depth research.

[0004] Platelet rich plasma (PRP) is a platelet concentrate obtained by centrifugation of peripheral blood. Its main components are platelets, fibrin and leukocytes. At the same time, platelets release a variety of growth factors after activation. The growth factors secreted by platelets include platelet-derived growth factor (PDGF), epidermal growth factor (EGF), insulin-like growth factor 1 (IGF1), transforming growth factor-β (TGF-β) and vascular endothelial growth factor (VEGF). These growth factors are the main components of PRP and play a very important role in tissue regeneration, vascular remodeling, angiogenesis, inflammatory response and other processes.

[0005] PRP can promote the proliferation of endometrial cells and promote angiogenesis by inducing the migration, proliferation and differentiation of endothelial cells. Matrix metalloproteinases (MMPs) participate in tissue regeneration and wound healing through extracellular matrix (ECM) degradation and wound remodeling. There are a variety of growth factors known to activate MMPs in PRP. PRP was co-cultured with endometrial stromal cells, and it was found that PPP regulated the expression of matrix-degrading enzymes MMP1, MMP3, MMP7 and MMP26 to a certain extent. Another animal study showed that PRP induced BMSCs IGF-1 expression and upregulated IL-10 expression by activating the NF-KB pathway, thereby enhancing the differentiation potential of BMSCs. PRP combined with BMSCs transplantation can significantly increase the thickness of rat endometrium and accelerate the repair and regeneration of damaged endometrium. In addition, by establishing an endometrial injury mouse model, the mice were euthanized 7 days after PRP treatment, and the cellular and molecular characteristics of mouse endometrial fibrosis and regeneration were evaluated by histological and immunofluorescence staining, as well as the measurement of fibrosis marker expression levels. The study found that the tissue sections of the group not receiving PRP treatment showed narrow endometrial cavity, atrophic columnar epithelium, tubular loss, and little matrix. Proliferating glands and endometrial stromal cells were observed in the PRP-treated group. Masson trichrome staining showed signs of endometrial fibrosis in the untreated group, while collagen deposition in the PRP-treated endometrium was significantly reduced. The expression of fibrosis-related factors (Collal, Tgfβ1, and Timp1) was reduced. Studies have shown that PRP promotes the recovery of endometrial structure and inhibits fibrosis after uterine horn injury. The inflammatory chemokines secreted by platelets after activation and the high concentration of leukocytes contained in PRP play an important role in inhibiting the body's inflammatory response and controlling infection. Studies have shown that compared with the control group, the expression level of the pro-inflammatory cytokine IL-1βmRNA in the PRP-treated group was significantly reduced, while c-Kit mRNA was upregulated. This shows that PRP can upregulate the expression level of anti-inflammatory factors and inhibit the release of inflammatory factors and excessive inflammatory response in the endometrium.

[0006] Although PRP has good therapeutic effects, the single drug treatment form is single and the efficacy is not good enough. Further development of better therapeutic drug compositions is needed. Summary of the invention

[0007] The purpose of the invention is to overcome the deficiencies in the prior art and provide a composition containing cryopreserved prp for use in treating gynecological diseases. The gynecological diseases described are specifically promoting the repair of endometrial damage, and more specifically promoting endometrial thickening.

[0008] More specifically, the cryopreserved PRP composition provided by the present invention contains PRP and a fertility-promoting peptide.

[0009] Specifically, the fertility-promoting peptide is prepared after separation and identification.

[0010] In order to solve the above technical problems, the present invention provides the following technical solutions:

[0011] The inventors found that rabbits are multiparous animals. They can reproduce 12 months a year. The gestation period is very short, generally about 32 days. They can reproduce continuously for many years, but the endometrium can still maintain a good state. Therefore, the inventors used the rabbit uterus as a research object to isolate and prepare a fertility-promoting peptide that promotes the proliferation of endometrial cells.

[0012] A reproductive peptide capable of promoting the proliferation of endometrial cells is prepared by freezing and crushing rabbit uterus to prepare a solution, enzymolyzing it with a neutral protease, and then ultrafiltration to obtain components with molecular weights of greater than 5KDa, greater than 3KDa and less than 5KDa, greater than 1KDa and less than 3KDa, and less than 1000Da, respectively, and identifying the effects of each component on promoting the proliferation of human endometrial stromal cells and rat human endometrial stromal cells, wherein the component greater than 1KDa and less than 3KDa has the strongest effect of promoting cell proliferation, collecting the components, and freeze-drying them to obtain polypeptide powder; adding the polypeptide obtained by ultrafiltration into dextran gel G-10, eluting with ultrapure water as an eluent after the sample completely penetrates into the gel, collecting the eluate, combining components with similar absorbances, and using a similar method as mentioned above to identify component No. 3 with a better effect of promoting the proliferation of human endometrial stromal cells. The component was further separated and purified using a DEAE-52 anion exchange chromatography column, and the active component that promoted cell proliferation was identified to be freeze-dried and mass spectrometry sequencing to identify a fertility-promoting peptide that promotes endometrial cell proliferation, whose amino acid sequence is shown in SEQ ID NO: 1.

[0013] The present invention further provides a modified peptide of the fertility-promoting peptide, wherein the modification is to make conservative substitutions of amino acids while still maintaining the activity of the polypeptide.

[0014] Specifically, the substitution is an amino acid sequence comprising or consisting of an amino acid sequence having one or more (preferably no more than 10, more preferably no more than 5, 4, 3, 2, 1) amino acid changes (preferably amino acid substitutions, more preferably conservative amino acid substitutions) compared to the amino acid sequence selected from SEQ ID NO: 1.

[0015] Furthermore, the present invention provides a kit containing cryopreserved prp, characterized in that the composition is composed of cryopreserved prp and fertility-promoting peptide; the cryopreserved prp is a freeze-dried powder prepared by freeze-drying the separated prp, which can be activated when used. The amino acid sequence of the fertility-promoting peptide is shown in SEQ ID NO: 1.

[0016] The present invention also provides a use of a fertility-promoting peptide in preparing a medicine for promoting endometrial repair.

[0017] Specifically, the medicine also contains a pharmaceutically acceptable carrier or excipient.

[0018] In the art, "pharmaceutically acceptable carrier" refers to a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material, formulation adjuvant or carrier conventionally used in the art, which together with the therapeutic agent constitutes a pharmaceutical composition administered to a subject. A pharmaceutically acceptable carrier is non-toxic to the recipient at the dosage and concentration used and is compatible with the other ingredients in the formulation. The pharmaceutically acceptable carrier is suitable for the drug being applied.

[0019] Unless otherwise expressly stated, the excipients used herein may refer to compounds used as or commonly referred to as fillers, diluents, bulking agents, etc., and it should be understood that these names are not exhaustive and they are not exclusive, for example, a specific excipient may be used as both a diluent and a filler. Unless otherwise expressly stated, the excipients used herein may refer to compounds used as or commonly referred to as binders, coatings, disintegrants, sweeteners, flavoring agents, and glidants.

[0020] Excipients can affect the stability, organoleptic properties and / or physical properties of a given drug. In a given active formulation, the weight ratio of active to excipient can be varied to alter the therapeutic, physical, appearance and / or organoleptic properties of the given formulation.

[0021] Suitable types of excipients for solid dosage forms include: additive binders, disintegrants, dispersants, fillers, sweeteners, glidants, flavoring agents, surfactants, wetting agents, preservatives and diluents. Although conventional pharmaceutical excipients can be used, they may not always function in exactly the same way as in conventional pharmaceutical processing.

[0022] Examples of suitable pharmaceutically acceptable excipients include one or more polymers, wetting agents or surfactants, pH adjusters, isotonicity adjusters, preservatives, buffers, and chelating agents, or any combination thereof.

[0023] Examples of suitable pharmaceutically acceptable pH adjusting agents include, but are not limited to, sodium hydroxide, citric acid, hydrochloric acid, boric acid, acetic acid, phosphoric acid, succinic acid, sodium hydroxide, potassium hydroxide, ammonium hydroxide, magnesium oxide, calcium carbonate, magnesium carbonate, magnesium aluminum silicate, malic acid, potassium citrate, sodium citrate, sodium phosphate, lactic acid, gluconic acid, tartaric acid, 1,2,3,4-butanetetracarboxylic acid, fumaric acid, diethanolamine, monoethanolamine, sodium carbonate, sodium bicarbonate, triethanolamine, or any combination thereof. In one embodiment, the pharmaceutically acceptable pH adjusting agent is present in an amount of about 0.01% to about 2.0% (weight / volume), preferably about 0.05% to about 1% (weight / volume).

[0024] Typical additives include: mono-, di- and polysaccharides; sugar alcohols and other polyols, for example, lactose, glucose, raffinose, melezitose, lactitol, maltitol, trehalose, sucrose, mannitol, starch, or a combination thereof; surfactants, for example, sorbitol, diphosphatidylcholine, or lecithin; etc. Typically, additives, such as fillers, are provided in amounts effective for the above purposes, usually from about 50% to about 99% by weight of the formulation.

[0025] Further, suitable carrier materials may be in the form of amorphous powder, crystalline powder or a combination of amorphous and crystalline powders. Suitable substances include carbohydrates, such as monosaccharides, such as fructose, galactose, glucose, D-mannose, sorbose, and the like; disaccharides, such as lactose, trehalose, cellobiose, and the like; cyclodextrins, such as 2-hydroxypropyl-cyclodextrin; and polysaccharides, such as raffinose, maltodextrin, dextran, and the like; amino acids, such as glycine, arginine, aspartic acid, glutamic acid, cysteine, lysine, and the like; organic salts prepared with organic acids and bases, such as sodium citrate, sodium ascorbate, magnesium gluconate, sodium gluconate, tromethamine hydrochloride, and the like; sugar alcohols, such as mannitol, xylitol, and the like. A preferred group of carriers includes lactose, trehalose, raffinose, maltodextrin, glycine, sodium citrate, tromethamine hydrochloride, and mannitol.

[0026] Further, the preservative can be selected from phenol, o-cresol, m-cresol, p-cresol, methylparaben, propylparaben, 2-phenoxyethanol, butylparaben, 2-phenylethanol, benzyl alcohol, chlorobutanol, and thiomerosal, bronopol, benzoic acid, imidazolinyl urea, chlorhexidine, sodium dehydroacetate, chlorocresol, ethylparaben, benzethonium chloride, chlorphenesine (3p-chlorophenoxypropane-1,2-diol) or its mixture. In a further aspect of the present invention, the concentration of the preservative is 0.1mg / ml to 20mg / ml. In a further aspect of the present invention, the concentration of the preservative is 0.1mg / ml to 5mg / ml. In a further aspect of the present invention, the concentration of the preservative is 5mg / ml to 10mg / ml. In a further aspect of the invention, the preservative is present in a concentration of 10 mg / ml to 20 mg / ml. Each of these specific preservatives constitutes an alternative aspect of the invention. The use of preservatives in medicaments is well known to the skilled person.

[0027] Beneficial effects of the present invention:

[0028] The fertility-promoting peptide provided by the present invention can significantly promote the proliferation and migration activity of human endometrial stromal cells, and can reduce the level of cell apoptosis, and has a good promotion effect. In addition, the present invention also prepares frozen PRP, which can significantly promote the repair of rat endometrial damage after being used in combination with the fertility-promoting peptide, and has a good therapeutic effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 Different concentrations of pro-fertility peptides can promote the proliferation of human endometrial stromal cells

[0030] Figure 2 Different concentrations of fertility-stimulating peptides can promote the migration of human endometrial stromal cells

[0031] Figure 3 Different concentrations of pro-fertility peptides can inhibit apoptosis of human endometrial stromal cells DETAILED DESCRIPTION

[0032] The specific embodiments of the present invention will be described in more detail below with reference to the accompanying drawings. Although the specific embodiments of the present invention are shown in the accompanying drawings, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided in order to enable a more thorough understanding of the present invention and to enable the scope of the present invention to be fully communicated to those skilled in the art.

[0033] Example 1 Activity verification of fertility-promoting peptides

[0034] The fertility promoting peptide shown in SEQ ID NO: 1 was commissioned to be artificially synthesized by Zhongke Huayao, and its purity was 99%. It was prepared into a mother solution with a concentration of 1 mg / mL for later use.

[0035] Human endometrial stromal cells (catalog number: bio-132385, Beijing Bio-Bowei Biotechnology Co., Ltd.) were placed in complete medium containing DMEM / F12, 10% FBS, and 1% double antibody at 37°C and 5% CO 2 Cultured in a medium incubator, digested with trypsin as is conventional in the art. After passaging, the CCK8 kit was used to detect cell growth according to the operating procedures in the instructions. The cells were passaged into a 96-well culture plate, and 0, 50, 100, 250, and 500 μg / mL of fertility-promoting peptides were added after 12 hours. The supernatant was removed, and DMEM / F12 culture medium containing 10% CCK8 was added to each well. The cells were incubated in an incubator for 2.5 hours, and the absorbance value was measured at 450 nm using an enzyme reader to determine the optimal concentration of the polypeptide. The blank control group was a complete culture medium without the addition of polypeptide. The results are shown in the figure. Figure 1 shown.

[0036] The results of adding 0, 50, 100, 250, 500 μg / mL fertility-promoting peptide to treat human endometrial stromal cells were Figure 1 The results showed that compared with the control group, gonadotropin promoted the proliferation of human endometrial stromal cells in a dose-dependent manner. At a concentration of 250 μg / mL, gonadotropin had the best effect on the proliferation of endometrial stromal cells.

[0037] Cell migration ability test: Human endometrial stromal cells were seeded in a 12-well plate with a plug. The composition of the culture medium was the same as that of the above groups. The culture medium was incubated at 37°C and 5% CO. 2 Culture overnight. When the cell density exceeds 90%, remove the plug. Take samples at 0, 6, 12, and 24 hours, take photos, and use ImageJ software to analyze the images and calculate the cell migration rate. Figure 2 shown.

[0038] Depend on Figure 2 It can be seen that after polypeptide treatment, with the increase of fertility-stimulating peptide concentration, the migration ability of cells in the 24th hour was gradually improved. When the concentration was 250 μg / mL, the cell migration rate reached a very significant 46.3%, which was significantly higher than the 25.7% of the control group (P<0.01), indicating that fertility-stimulating peptide can significantly improve cell activity.

[0039] At the same time, an equal amount of cells cultured in each group for 24 hours were washed three times with PBS, and then the tissue blocks were ground with liquid nitrogen. The tissue lysis solution was lysed on ice and centrifuged for 30 minutes, and the supernatant was taken as the total protein. Some samples were taken to determine the protein concentration using the BCA protein content assay kit. The samples were diluted to an appropriate concentration for SDS-PAGE electrophoresis, transferred to a PVDF membrane using the semi-dry transfer method, blocked for 1 hour with the primary antibody and incubated overnight at 4°C, and the labeled secondary antibody was added for incubation at room temperature. The membrane was scanned or photographed, and the net optical density of the bands was analyzed using the gel image processing system (ImageJ), photographed, and saved. The grayscale value of the Western blotting protein bands was analyzed using ImageJ software, and the relative protein expression of Casepase-3 apoptotic protein / β-actin was calculated. The results are shown in Figure 3 shown.

[0040] Depend on Figure 3 It can be seen that compared with the control group, the expression of Caspase-3 protein was significantly reduced after cells were treated with peptides at various concentrations (P<0.05). Since Caspase-3 is an effector protease closely related to cell apoptosis, Caspase-3 can be used as an indicator of cell apoptosis. Figure 3 It can be seen that fertility-stimulating peptide can also effectively reduce cell apoptosis.

[0041] Example 2 Preparation of frozen prp

[0042] Preparation of PRP: After the rats were anesthetized, blood was collected through the abdominal vena cava. A total of 10 mL of venous blood from multiple rats was mixed with anticoagulants and centrifuged at 3000 r / min for 5 minutes to collect the plasma layer and the buffy coat. Then, the blood was centrifuged at 3200 r / min for 5 minutes to obtain 5.8 mL of PRP.

[0043] Mix 5.8 mL of PRP with 5.8 mL of freeze-drying protective agent (0.5 M trehalose + 10% glycerol), freeze at -40°C, put into a freeze dryer pre-cooled to -46°C for vacuum drying, keep the vacuum degree at <15 Pa, and take out after drying for 12 hours to obtain the frozen prp.

[0044] Example 3 Animal experiment on cryopreservation of prp and fertility-promoting peptide

[0045] The frozen PRP was fully mixed and rehydrated with sterile water for injection to adjust the concentration to 1 mg / mL. Then, 100 U of thrombin powder and 1 mL of calcium gluconate were added to 1 mL and mixed to obtain activated PRP.

[0046] Model preparation: The model was prepared using a mature method in this field. Specifically, female SD rats in estrus were intraperitoneally injected with 4% chloral hydrate (0.8 mL / 100 g). After the corneal reflex and righting reflex disappeared, the back surgical area was prepared, disinfected locally with iodine, and placed on a sterile operating table. Through a longitudinal incision, the uterus was separated layer by layer to expose one side. The "0" line was used to ligate the two ends near the ovary and the vagina. The gauze was soaked with isotonic saline and placed near the incision. The toothless forceps straightened the uterus. A 1m syringe was slowly injected with 95% ethanol by volume from the distal end of the ovary. The uterine cavity was filled. After the needle was kept in the uterine cavity for 5 minutes, the remaining ethanol was sucked out, and the uterine cavity was repeatedly rinsed with isotonic saline for 3 times. After confirming that there was no bleeding and fluid residue, the uterus was returned to the abdominal cavity, and then sutured layer by layer, and the epidermal suture was disinfected with iodine. Penicillin (80,000 U / rat) was continuously injected intramuscularly for 3 days after surgery to prevent infection. Pay attention to keeping the rats warm after surgery and send them back to the animal room after waking up.

[0047] The experimental groups were 10 rats in each group. The normal group was not intervened and raised normally. The model group rats were injected with 0.2 mL of PBS through the uterine cavity after modeling; the fertility peptide treatment group: the fertility peptide of the present invention was injected with 0.2 mL (1 mg / mL) through the uterine cavity after modeling; the cryopreserved prp treatment group: the activated PRP was injected with 0.2 mL through the uterine cavity after modeling; the fertility peptide combined with cryopreserved prp treatment group (combined treatment group): the fertility peptide of the present invention was injected with 0.2 mL (1 mg / mL) and activated PRP 0.2 mL through the uterine cavity after modeling; the positive control group: the estrogen was injected with 0.2 mL (1 mg / mL) through the uterine cavity after modeling. Each group was administered once every 4 days. After the intervention of 4 estrous cycles, the rats were killed by cervical dislocation at the estrus period, the lower abdominal hair was shaved, the local alcohol disinfection was carried out, the skin was cut open, and the uterus was separated. The rats were fixed with 4% paraformaldehyde, embedded in paraffin, and stained with HE. The uterine tissue morphology of each group of rats was observed under an optical microscope, and the thickness of the endometrium and the number of blood vessels and glands were measured. The results are shown in Table 1.

[0048] Table 1 Results of endometrial thickness, blood vessels and gland numbers in each group of rats

[0049] Group Endometrial thickness (μm) Number of blood vessels Number of glands Normal group 415.84±58.34 4.18±0.46 2.51±0.30 Model Group 279.55±41.87* 1.64±0.25* 0.96±0.17* Fertility-stimulating peptide treatment group 389.17±38.43## 3.25±0.29## 1.94±0.22# Cryopreserved prp treatment group 355.19±29.41# 3.04±0.18## 1.82±0.13# Combination therapy group 405.76±48.79## 4.12±0.27## 2.43±0.28# Positive control group 340.56±24.18# 2.86±0.14# 1.74±0.15

[0050] *Compared with the normal group, *P<0.01; #Compared with the model group, #<0.05; ##<0.01.

[0051] HE staining results of endometrium in rats in each group showed that the uterus and uterine cavity of normal rats during estrus were intact, the endometrium was in a normal wavy state, the number of blood vessels and glands was large, and the glandular lumen was dilated. The uterus and uterine cavity of rats in the model group during estrus were significantly enlarged, the endometrium was thinned, the glands and blood vessels were sparse, and the glandular cavity was smaller. After treatment, the endometrial state of each treatment group was well improved, especially in the uterus of rats in the combined treatment group, most of the complete uterine cavity structure was visible, the endometrial surface had a certain wavy shape, the thickness was significantly thickened, the glands were also significantly increased, and there were more new capillaries. This shows that the combined use of fertility-promoting peptides and cryopreserved prp can significantly improve the treatment of endometrial damage and has a significant synergistic effect.

[0052] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. Although embodiments of the present invention have been shown and described, those skilled in the art will appreciate that various changes, modifications, substitutions and variations may be made to these embodiments without departing from the principles and purpose of the present invention, and that the scope of the present invention is defined by the claims and their equivalents.

[0053] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents. However, these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A reproductive peptide that promotes the proliferation of endometrial cells, characterized in that The amino acid sequence is shown in SEQ ID NO:

1.

2. Use of the fertility-promoting peptide for promoting endometrial cell proliferation as claimed in claim 1 in the preparation of a drug for promoting the repair of endometrial damage.

3. Use of frozen prp and the fertility-promoting peptide described in claim 1 in the preparation of a medicine kit for promoting the repair of endometrial damage, wherein the frozen prp is prepared by mixing the separated prp with the freeze-drying protective agent 0.5M trehalose + 10% glycerol in a volume ratio of 1:1, and then vacuum drying to obtain the frozen prp; when the frozen prp is used, thrombin powder and calcium gluconate are added to activate it, and the activated state prp is obtained, which can be injected for use.

4. The use according to claim 3, characterized in that The prp is purchased commercially or obtained directly from blood by centrifugation.

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