Use of a composition containing cryopreserved PRP in the treatment of gynecological diseases

Abstract

The present invention provides the use of a composition containing cryopreserved PRP in the treatment of gynecological diseases. It is also provided that the reproductive promoting peptide can significantly promote the proliferation and migration activities of human endometrial stromal cells, and can reduce the apoptosis level of cells, showing a good promoting effect. In addition, the present invention has also prepared cryopreserved PRP, and after combining it with the reproductive promoting peptide, it can significantly promote the repair of rat endometrial injury, showing a good therapeutic effect.

Description

Technical Field

[0001] This 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 for nurturing life and a key component of female reproductive health. When the endometrium is severely damaged, such as in pathological conditions like repeated curettage and infection, the basal layer of the uterus is damaged, and the proliferative ability of basal layer cells is impaired. The most direct manifestation is the thinning of the endometrium. At the same time, during the repair process of severely damaged endometrium, pathological repair may occur, resulting in varying degrees of fibrosis, and in severe cases, even intrauterine adhesions leading to uterine cavity occlusion. Endometrial damage easily causes menstrual reduction, amenorrhea, repeated miscarriages, and infertility, and may manifest as repeated implantation failure (RIF) during in vitro fertilization-embryo transfer. When only the functional layer of the endometrium is mildly damaged, the epithelial cells on the wound surface can quickly achieve regeneration, and the endometrium can be repaired without scarring. However, 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 endometrial stem cells in the basal layer cannot proliferate and regenerate. On the other hand, the newly formed scar tissue usually has no vascular structure, and the insufficient blood supply further hinders the regeneration and repair of the endometrium. Therefore, it is necessary to reduce the formation of fibrous scars and promote the regeneration and repair of damaged endometrium to restore the uterine cavity structure and the normal physiological function of the endometrium.

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

[0004] Platelet-rich plasma (PRP) is a platelet concentrate obtained by centrifuging peripheral blood. Its main components are platelets, fibrin, and white blood cells. At the same time, after platelet activation, a variety of growth factors are released. 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 processes such as tissue regeneration, blood vessel remodeling, angiogenesis, and inflammatory responses.

[0005] PRP can promote the proliferation of endometrial cells and promote neovascularization by inducing the migration, proliferation, and differentiation of vascular endothelial cells. Matrix metalloproteinases (MMPs) are involved in tissue regeneration and wound healing through extracellular matrix (ECM) degradation and wound remodeling. There are multiple known growth factors in PRP that can activate MMPs. Co-culturing PRP with endometrial stromal cells revealed that PPP upregulated the expression of matrix-degrading enzymes MMP1, MMP3, MMP7, and MMP26 to a certain extent. Another animal study showed that PRP induced the expression of IGF-1 in BMSCs and upregulated the expression of IL-10 by activating the NF-κB pathway, thereby enhancing the differentiation potential of BMSCs. The transplantation of PRP combined with BMSCs could significantly increase the thickness of the endometrium in rats and accelerate the repair and regeneration of damaged endometrium. In addition, by establishing a mouse model of endometrial injury, the mice were euthanized 7 days after PRP treatment, and histological and immunofluorescence staining, as well as measurement of the expression levels of fibrosis markers, were used to evaluate the cellular and molecular characteristics of endometrial fibrosis and regeneration in mice. It was found that the tissue sections of the group not receiving PRP treatment showed a narrow endometrial cavity, atrophic columnar epithelium, loss of the lumen, and very little stroma. Proliferating glands and endometrial stromal cells were observed in the PRP-treated group. Masson's trichrome staining in the untreated group showed signs of endometrial fibrosis, while the deposition of collagen in the PRP-treated endometrium was significantly reduced. The expression of fibrosis-related factors (Collal, Tgfβ1, and Timp1) decreased. The study showed that PRP promoted the restoration of the endometrial structure and inhibited fibrosis after uterine horn injury. The inflammatory chemokines secreted after platelet activation in PRP and the high concentration of white blood cells contained in it play an important role in inhibiting the body's inflammatory response and controlling infection. The study showed that compared with the control group, the expression level of the pro-inflammatory cytokine IL-1β mRNA in the PRP-treated group was significantly reduced, while c-Kit mRNA was upregulated. This indicates that PRP can upregulate the expression level of anti-inflammatory factors and inhibit the release of inflammatory factors and excessive inflammatory responses in the endometrium.

[0006] Although PRP has good therapeutic effects, the form of a single drug treatment is single, and the curative effect is not good enough. There is a need to further develop a drug composition with better therapeutic effects. Summary of the Invention

[0007] The object of the invention is to overcome the deficiencies in the prior art and provide the use of a composition containing cryopreserved prp in the treatment of gynecological diseases. The specific gynecological disease is to promote the repair of endometrial injury, and more specifically, to promote endometrial thickening.

[0008] More specifically, the cryopreserved prp composition provided by the present invention contains prp and reproductive peptides.

[0009] Specifically, the reproductive promoting peptide is prepared after isolation and identification.

[0010] To solve the above technical problems, the present invention provides the following technical solutions:

[0011] The inventors have found through research that rabbits are polyovulatory animals. They can reproduce throughout the 12 months of a year, with a very short pregnancy period, generally around 32 days, and can reproduce continuously for many years, yet the endometrium can still maintain a good state. Therefore, the inventors used rabbit uterus as the research object and isolated and prepared a reproductive promoting peptide with the effect of promoting the proliferation of endometrial cells.

[0012] A reproductive promoting peptide with the effect of promoting the proliferation of endometrial cells is prepared by freezing and pulverizing a rabbit uterus to form a solution, enzymatically digesting it with neutral protease, and then obtaining components with molecular weights greater than 5KDa, greater than 3KDa and less than 5KDa, greater than 1KDa and less than 3KDa, and less than 1000Da through ultrafiltration. The proliferation promoting effects of each component on human endometrial stromal cells and rat human endometrial stromal cells are respectively identified. Among them, the component with a molecular weight greater than 1KDa and less than 3KDa has the strongest effect on promoting cell proliferation. This component is collected and freeze-dried to obtain a polypeptide powder; the polypeptide prepared by ultrafiltration is added to Sephadex G-10. After the sample completely penetrates into the gel, ultrapure water is used as the eluent for elution. The eluate is collected, and the components with similar absorbance are combined and then identified by a similar method as described above to obtain Component 3 with a good effect on promoting the proliferation of human endometrial stromal cells. This component is further separated and purified using a DEAE-52 anion exchange chromatography column, and the active component with a good effect on promoting cell proliferation is identified and freeze-dried, and then subjected to mass spectrometry sequencing to identify a reproductive promoting peptide with the effect of promoting the proliferation of endometrial cells, and its amino acid sequence is as shown in SEQ ID NO: 1.

[0013] Furthermore, the present invention provides a modified peptide of the reproductive promoting peptide, and the modification is to conservatively substitute amino acids while still maintaining the activity of the polypeptide.

[0014] Specifically, the substitution includes an amino acid sequence having one or more (preferably not exceeding 10, more preferably not exceeding 5, 4, 3, 2, 1) amino acid changes (preferably amino acid substitution, more preferably amino acid conservative substitution) compared with the amino acid sequence selected from SEQ ID NO: 1 or consisting of the amino acid sequence.

[0015] Furthermore, the present invention provides a medicine box containing cryopreserved PRP, characterized in that the composition is composed of cryopreserved PRP and reproductive promoting peptides; the cryopreserved PRP is a freeze-dried powder prepared by freeze-drying the separated PRP, and can be activated during use. The reproductive promoting peptides have the amino acid sequence shown in SEQ ID NO: 1.

[0016] The present invention also provides the use of reproductive promoting peptides in the preparation of drugs for promoting endometrial repair.

[0017] Specifically, the drug further contains a pharmaceutically acceptable carrier or excipient.

[0018] In the art, "pharmaceutically acceptable carrier" refers to non-toxic solid, semi-solid or liquid fillers, diluents, encapsulating materials, formulation aids or carriers conventionally used in the art, which together with a therapeutic agent form a pharmaceutical composition administered to a subject. The pharmaceutically acceptable carrier is non-toxic to the recipient at the applied doses and concentrations and is compatible with the other components in the formulation. The pharmaceutically acceptable carrier is suitable for the applied drug.

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

[0020] Excipients can affect the stability, sensory properties and / or physical properties of the administered drug. In the administered active agent formulation, the weight ratio of the active agent to the excipient can vary to alter the therapeutic, physical, appearance and / or sensory properties of the administered 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 act in exactly the same way as in traditional pharmaceutical processing.

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

[0023] Examples of suitable pharmaceutically acceptable pH adjusters 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 adjuster is present in an amount of from about 0.01% to about 2.0% (weight / volume), preferably from about 0.05% to about 1% (weight / volume).

[0024] Typical additives include: mono-, di- and polysaccharides; sugar alcohols and other polyols, e.g., lactose, glucose, raffinose, melezitose, lactitol, maltitol, trehalose, sucrose, mannitol, starch, or combinations thereof; surfactants, e.g., sorbitol, dipalmitoyl phosphatidylcholine, or lecithin; and the like. Typically, additives, such as fillers, are provided in an amount effective for the above purposes, often about 50% to about 99% of the weight of the formulation.

[0025] Further, suitable carrier materials can be in the form of amorphous powders, crystalline powders, or combinations of amorphous and crystalline powders. Suitable substances include carbohydrates, such as monosaccharides, e.g., fructose, galactose, glucose, D-mannose, sorbose, etc.; disaccharides, e.g., lactose, trehalose, cellobiose, etc.; cyclodextrins, e.g., 2-hydroxypropyl-cyclodextrin; and polysaccharides, e.g., raffinose, maltodextrin, dextran, etc.; amino acids, e.g., glycine, arginine, aspartic acid, glutamic acid, cysteine, lysine, etc.; organic salts prepared with organic acids and bases, e.g., sodium citrate, sodium ascorbate, magnesium gluconate, sodium gluconate, tromethamine hydrochloride, etc.; sugar alcohols, e.g., mannitol, xylitol, etc. 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, methyl p-hydroxybenzoate, propyl p-hydroxybenzoate, 2-phenoxyethanol, butyl p-hydroxybenzoate, 2-phenylethanol, benzyl alcohol, chlorobutanol, and thiomerosal, bronopol, benzoic acid, imidurea, chlorhexidine digluconate, sodium dehydroacetate, parachlorometacresol, ethyl p-hydroxybenzoate, benzethonium chloride, chlorphenesine (3-p-chlorophenoxypropane-1,2-diol) or a mixture thereof. In a further aspect of the present invention, the concentration of the preservative present is from 0.1 mg / ml to 20 mg / ml. In a further aspect of the present invention, the concentration of the preservative present is from 0.1 mg / ml to 5 mg / ml. In a further aspect of the present invention, the concentration of the preservative present is from 5 mg / ml to 10 mg / ml. In a further aspect of the present invention, the concentration of the preservative present is from 10 mg / ml to 20 mg / ml. Each of these specific preservatives constitutes an alternative aspect of the present invention. The use of preservatives in pharmaceuticals is well known to those skilled in the art.

[0027] Advantages of the present invention:

[0028] The reproductive promoting peptide provided by the present invention can significantly promote the proliferation and migration activity of human endometrial stromal cells, and can reduce the apoptosis level of the cells, having a good promoting effect. In addition, the present invention also prepares cryopreserved prp, and after combining it with the reproductive promoting peptide, it can significantly promote the repair of rat endometrial injury, having a good therapeutic effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 Reproductive promoting peptides at different concentrations can promote the proliferation of human endometrial stromal cells

[0030] Figure 2 Reproductive promoting peptides at different concentrations can promote the migration of human endometrial stromal cells

[0031] Figure 3 Reproductive promoting peptides at different concentrations can inhibit the apoptosis of human endometrial stromal cells DETAILED DESCRIPTION OF THE EMBODIMENTS

[0032] The specific embodiments of the present invention will be described in more detail with reference to the accompanying drawings. Although the specific embodiments of the present invention are shown in the 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 so that the present invention can be more thoroughly understood and the scope of the present invention can be fully conveyed to those skilled in the art.

[0033] Example 1 Verification of the activity of the reproductive promoting peptide

[0034] The reproductive promoting peptide shown in SEQ ID NO: 1 was entrusted to Zhongke Huayao for artificial synthesis, and its purity was 99%. It was prepared into a stock solution with a concentration of 1 mg / mL for standby.

[0035] Human endometrial stromal cells, product number: bio-132385, Beijing BioWin Biotechnology Co., Ltd., were cultured in a complete medium of DMEM / F12, 10% FBS, and 1% double antibody in an incubator at 37°C and 5% CO2, and were digested with trypsin routinely used in the art. After passage, a CCK8 kit was used to detect the cell growth according to the operation steps of the instruction manual. The cells were passaged into a 96-well culture plate. After 12 h, 0, 50, 100, 250, 500 μg / mL of the reproductive promoting peptide were added. The supernatant was removed, and a DMEM / F12 culture solution containing 10% CCK8 was added to each well. The plate was incubated in the incubator for 2.5 h. The absorbance value was measured at 450 nm using an enzyme-linked immunosorbent assay (ELISA) reader to determine the optimal concentration of the polypeptide. The blank control group was a complete medium without the addition of the polypeptide. The results were as Figure 1 shown.

[0036] The results of treating human endometrial stromal cells with 0, 50, 100, 250, 500 μg / mL of the reproductive promoting peptide were Figure 1 shown. Compared with the control group, the reproductive promoting peptide had a dose-dependent promotion of the proliferation of human endometrial stromal cells. At a concentration of 250 μg / mL, the reproductive promoting peptide had the best effect on the proliferation of endometrial stromal cells.

[0037] Detection of cell migration ability: Human endometrial stromal cells were seeded in a 12-well plate with inserts. The components of the culture medium were the same as those in the previous groups. The cells were cultured overnight at 37°C and 5% CO2. When the cell density exceeded 90%, the inserts were removed. Samples were taken at 0, 6, 12, and 24 h, photographed, and ImageJ software was used for image analysis to calculate the cell migration rate. The results were as Figure 2 shown.

[0038] As Figure 2 can be seen, after treatment with the polypeptide, with the increase in the concentration of the reproductive promoting peptide, the migration ability of the cells at 24 h was gradually improved. When the concentration was 250 μg / mL, the cell migration rate reached a very significant 46.3%, significantly higher than 25.7% of the control group (P < 0.01), indicating that the reproductive promoting peptide could significantly improve the cell activity.

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

[0040] As Figure 3 can be seen, compared with the control group, after the cells were treated with polypeptides at various concentrations, the expression of Casepase-3 protein was significantly reduced (P<0.05). Since Caspase-3 is an effector protease closely related to cell apoptosis and can be used as an indicator of cell apoptosis, therefore, Figure 3 it can be seen that the reproductive promoting peptide can also effectively reduce cell apoptosis.

[0041] Example 2 Preparation of cryopreserved PRP

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

[0043] After mixing 5.8 mL of PRP with 5.8 mL of cryoprotectant (0.5 M trehalose + 10% glycerol), it was cryogenically frozen at -40 °C and placed in a freeze dryer pre-cooled to -46 °C for vacuum drying. The vacuum degree was maintained at <15 Pa, and after drying for 12 h, it was taken out, which was the cryopreserved PRP.

[0044] Example 3 Animal experiment of cryopreserved PRP and reproductive promoting peptide

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

[0046] Model preparation: It was prepared by a mature method in the art. Specifically, female SD rats in estrus were taken and intraperitoneally injected with 4% chloral hydrate (0.8 mL / 100 g). After the corneal reflex and righting reflex disappeared, the back surgical area was shaved, locally disinfected with iodophor, and placed on a sterile operating table. Through a longitudinal incision, layer by layer separation was performed to expose one side of the uterus. The two ends near the ovary and near the vagina were ligated with "0" suture. The gauze was moistened with isotonic saline and padded near the incision. The uterus was straightened with an atraumatic forceps. A 1 mL syringe was slowly injected with 95% ethanol by volume from the distal end of the ovary until the uterine cavity was filled. The needle was kept in the uterine cavity for 5 minutes, and then the remaining ethanol was aspirated back. Then the uterine cavity was repeatedly rinsed with isotonic saline 3 times. After confirming no bleeding and no liquid residue, the uterus was returned to the abdominal cavity, and then sutured layer by layer. The epidermal suture was disinfected with iodophor. After the operation, penicillin (80,000 U / rat) was continuously intramuscularly injected for 3 days to prevent infection. After the operation, the rats were kept warm, and after waking up, they were sent back to the animal house.

[0047] The experimental groups were as follows: Each group had 10 rats. The normal group was not intervened and was normally raised. The rats in the model group were intrauterinely injected with 0.2 mL of PBS after modeling; the reproductive peptide treatment group: 0.2 mL (1 mg / mL) of the reproductive peptide of the present invention was intrauterinely injected after modeling; the cryopreserved PRP treatment group: 0.2 mL of activated PRP was intrauterinely injected after modeling; the reproductive peptide combined with cryopreserved PRP treatment group (combined treatment group): 0.2 mL (1 mg / mL) of the reproductive peptide of the present invention and 0.2 mL of activated PRP were intrauterinely injected after modeling; the positive control group: 0.2 mL (1 mg / mL) of estrogen was intrauterinely injected after modeling. Each group was administered once every 4 days. After 4 estrous cycles of intervention, the rats were sacrificed by decapitation in estrus. The abdominal hair of the rats was shaved, locally disinfected with alcohol, the skin was cut open, and the uterus was separated. Fixed with 4% paraformaldehyde, embedded in paraffin, stained with HE, and the morphological changes of the uterine tissues of rats in each group were observed with an optical microscope, and the endometrial thickness 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 of rats in each group

[0049] Group Endometrial thickness (μm) Number of blood vessels (pcs) Number of glands (pcs) 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* Reproductive promoting 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# Combined treatment 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 the endometrium of rats in each group: During the estrus period of normal rats, the uterine cavity structure was intact, the endometrium was in a normal wavy state, with a large number of blood vessels and glands, and the glandular lumen was dilated. During the estrus period of rats in the model group, the uterine cavity was significantly enlarged, the endometrium was thinned, the glands and blood vessels were sparse, and the glandular lumen became smaller. After treatment in each treatment group, the state of the endometrium was improved to a certain extent. Especially in the combined treatment group, most of the intact uterine cavity structure could be seen in the rats' uterus, the surface of the endometrium had a certain wavy shape, the thickness was significantly increased, the glands were also significantly increased, and there were more newly formed capillaries. This indicates that the combined use of reproductive peptide and cryopreserved PRP can significantly improve the treatment of endometrial injury and has a significant synergistic effect.

[0052] In the description of this specification, the descriptions referring to terms such as "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. mean that the specific features, structures, materials or characteristics described in connection 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 can be combined in a suitable manner in any one or more embodiments or examples. Although the embodiments of the present invention have been shown and described, those of ordinary skill in the art can understand that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and purposes of the present invention, and 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 and are not intended to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent substitutions for some or all of the technical features; and these modifications or substitutions 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 platelet rich plasma (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 separated PRP with a lyophilizing agent in a volume ratio of 1:1 and then vacuum drying to obtain the frozen PRP, wherein the lyophilizing agent is 0.5M trehalose + 10% glycerol; when used, the frozen PRP is activated by adding thrombin powder and calcium gluconate to obtain activated PRP, 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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