Composition for treating gynecological diseases and preparation method and application thereof

Abstract

The present invention discloses a composition for treating gynecological diseases, a preparation method and an application thereof, and specifically relates to the field of medical technology. The composition comprises placental mesenchymal stem cells (PMSCs) extract, vitamin C, cyclosporine, an amino acid mixture and sodium hyaluronate. The composition of the present invention utilizes a multi-factor secretion extraction mixture to perform rehabilitation treatment on cervicitis and cervical erosion, that is, utilizes factors to regulate and repair inflammation, does not require customized treatment, and can meet the needs of almost all patients, even for patients with multiple underlying diseases, without worrying about adverse reactions after use.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a composition for treating gynecological diseases, a preparation method and application thereof. Background Art

[0002] Cervicitis and cervical erosion are common gynecological diseases that have a certain impact on women's reproductive health. For these two diseases, the treatment options mainly include general treatment and prevention, drug therapy, physical therapy, surgical treatment, Chinese medicine-assisted treatment, and regular examination and monitoring. These treatment options will be introduced in detail below.

[0003] The existing drug treatments mainly include antibiotics: for cervicitis, appropriate antibiotics can be selected according to the pathogen to eliminate the source of infection. Anti-inflammatory drugs: used to relieve the pain and discomfort caused by cervicitis and reduce the inflammatory response. Local medications: such as cervical suppositories, lotions, etc., can directly act on the lesion site, reduce inflammation, and promote the repair of the erosion surface.

[0004] Physical therapy mainly includes cryotherapy: using the freezing principle to make the diseased tissue necrotize and fall off, and promote the growth of new tissue. Laser therapy: using the energy of laser to act on the lesion to eliminate inflammation and promote the healing of erosion surface. Electric ironing therapy: using electric ironing to make the diseased tissue coagulate and necrotize to achieve the purpose of treatment.

[0005] Surgical treatment is mainly for patients with severe cervicitis and cervical erosion who are not responsive to drug treatment. Surgical treatment can be considered. Surgical methods include cervical conization, cervical circumcision, etc. The specific choice should be determined according to the patient's condition.

[0006] Although there are many treatment options for cervicitis and cervical erosion, each treatment method has certain defects and shortcomings. The following are the inevitable defects of existing treatment options:

[0007] First, although drug treatment is relatively simple, the treatment time is long and the effect may vary from person to person. Drugs need to be absorbed and metabolized by the body before they can act on the lesion, so the treatment process is relatively slow. At the same time, drug treatment also has certain side effects and drug resistance risks. Long-term use may cause the body to resist the drug and reduce the treatment effect. Secondly, although physical therapy such as freezing, laser and electric ironing can directly act on the lesion, the treatment process may cause certain pain and discomfort. In addition, physical therapy may have limited effect on patients with larger lesions or deeper inflammatory infiltration, and there is a risk of recurrence after treatment. Finally, surgical treatment is usually for patients with more serious conditions or ineffective drug treatment. Although it can effectively remove the lesion tissue, the surgical trauma is large and the recovery time is long. At the same time, surgical treatment may affect the patient's fertility, which is a major defect for patients with fertility needs.

[0008] In summary, the existing treatments for cervicitis and cervical erosion each have their own advantages and disadvantages, so finding a treatment that is convenient and has few side effects is an urgent problem to be solved. Summary of the invention

[0009] To this end, the present invention provides a composition for treating gynecological diseases and a preparation method and application thereof to solve the above-mentioned problems.

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

[0011] According to the first aspect of the present invention, a composition for treating gynecological diseases is provided, characterized in that the composition comprises placental mesenchymal stem cells (PMSCs) extract, vitamin C, cyclosporine, an amino acid mixture, and sodium hyaluronate.

[0012] Furthermore, the composition comprises 4-8 mL of placental mesenchymal stem cell (PMSCs) extract, 10-100 μl / mL of vitamin C, 5-45 μg / mL of cyclosporine, 2-25 mg / mL of amino acid mixture, and 0.01-0.5 mg / ml of sodium hyaluronate.

[0013] Furthermore, the preparation method of the placental mesenchymal stem cell PMSCs extract includes: culturing the placental mesenchymal stem cells using α-MEM culture medium, the growth density is not less than 80%, the cell growth state is good, and the cell viability is >95%, removing the supernatant and adding an induction drug, continuing the induction culture, and after the culture is completed, collecting the cytokines secreted by the cells and purifying them to obtain the placental mesenchymal stem cell PMSCs extract.

[0014] Furthermore, the induction drugs include adenosine diphosphate heptose, protein kinase, vitamin E and dendrobium polysaccharide.

[0015] Furthermore, the induction drugs include 5-15 μL / mL of adenosine diphosphate heptose, 3-10 ng / mL of protein kinase, 5-20 μL / mL of vitamin E and 5-15 μL / mL of dendrobium polysaccharide.

[0016] Adenosine diphosphate heptose (CAS: 7758-11-4):

[0017] After entering the host cell, ADP-heptose binds to the N-terminal domain of the kinase ALPK1 and stimulates the C-terminal kinase domain to phosphorylate and activate TIFA, mediating the activation of NF-KB and the production of cytokines that depend on the type I secretion system.

[0018] Protein kinase (CAS:126643-37-6):

[0019] Protein kinases (PKs) are the largest group in the kinase family. They can affect the structure and activity of substrates by catalyzing the phosphorylation of specific substrate proteins (transferring the γ-phosphate group at the end of ATP to specific amino acids in substrate proteins), and then participate in a series of cell signal transduction and regulation processes, thus playing an important role in cell growth, metabolism, differentiation and apoptosis. Protein kinases are associated with many diseases, including cancer, diabetes and inflammation. Therefore, targeted inhibition of protein kinases has become an attractive therapeutic strategy for the treatment of related diseases.

[0020] Vitamin E:

[0021] 1. Antioxidant Effect

[0022] Vitamin E is an important antioxidant that prevents free radicals from damaging human cells. Free radicals are products of metabolic processes and environmental factors (such as radiation and pollutants). They damage cells and tissues, thereby inducing oxidative stress. Vitamin E can protect cells from damage by inhibiting the production of free radicals and relieving oxidative stress.

[0023] 2. Strengthen the immune system

[0024] Vitamin E can enhance the function of the immune system and improve the body's ability to fight infection. Studies have shown that vitamin E can increase the activity and number of lymphocytes, promote the synthesis of immunoglobulins, and thus enhance the body's resistance. In addition, vitamin E can also reduce inflammatory responses, improve the interaction between immune cells, and enhance the immune system's resistance to disease.

[0025] Dendrobium polysaccharide product number QX-0525-070:

[0026] 1. Regulate immune function:

[0027] The human body's immune regulation is achieved by the combined action of multiple active polysaccharides, which is the basis for polysaccharides to exert their physiological and pharmacological effects.

[0028] 2. Anti-oxidation and anti-aging:

[0029] It can improve the activity of antioxidant enzymes and inhibit the activity of lipid peroxidase, thereby protecting biological membranes and delaying aging. Dendrobium contains highly active superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), and has strong antioxidant capacity. In alkaline conditions, Dendrobium polysaccharides have good antioxidant activity.

[0030] According to a second aspect of the present invention, a method for preparing a composition for treating gynecological diseases is provided, the method comprising mixing raw materials to obtain the composition.

[0031] According to the third aspect of the present invention, a composition for treating gynecological diseases is provided for use in preparing a composition for treating cervicitis.

[0032] According to a fourth aspect of the present invention, a composition for treating gynecological diseases is provided for use in preparing a composition for treating cervical erosion.

[0033] Function of each component:

[0034] Placenta mesenchymal stem cell supernatant extract:

[0035] PMSCs, or placental mesenchymal stem cells, have multidirectional differentiation potential and tissue repair ability. They can differentiate into various cell types, such as fibroblasts, endothelial cells, smooth muscle cells, etc., thereby promoting the repair and regeneration of damaged tissues. They can also secrete a variety of cytokines and growth factors, such as inflammatory factors, transforming growth factor-β (TGF-β), epidermal growth factor (EGF), etc. These factors can inhibit inflammatory responses, reduce tissue damage, and promote tissue repair. They can also secrete angiogenic factors (VEGF), etc., to promote new blood vessel formation, improve local blood circulation, and increase the supply of oxygen and nutrients, thereby promoting tissue repair and regeneration. PMSCs can also regulate immune function, inhibit immune response, reduce inflammatory response, reduce the degree of tissue damage, and promote tissue repair.

[0036] Vitamin C:

[0037] It is an effective antioxidant that helps neutralize free radicals and reduce damage to cells caused by oxidative stress. It is also an essential nutrient for the synthesis of collagen, which is an important component of connective tissue such as skin, bones, and blood vessels, and helps maintain skin elasticity.

[0038] Cyclosporine (CAS: 59865-13-3):

[0039] It is an immunosuppressant that is mainly used to prevent rejection reactions after organ transplantation. It can also be used in cell transplantation to reduce immune rejection reactions between cells and tissues. It also has a certain anti-inflammatory effect and can reduce inflammatory reactions.

[0040] Amino Acid Mixture (Cat. No.: GBW(E)100062):

[0041] The importance of amino acids to the body: Amino acids are not only important components of cells, but also key to the normal functioning of the body. Amino acids play an important role in energy metabolism. They can be converted into ATP to provide energy for cells. In addition, amino acids can also regulate the body's acid-base balance and osmotic pressure to maintain the body's internal environment. Adequate supply of amino acids is essential for the body's normal physiological functions.

[0042] Anti-inflammatory effect of amino acid supplementation: Another important role of amino acids is their anti-inflammatory effect. In inflammatory reactions, amino acids can stimulate the immune system to produce anti-inflammatory molecules, such as interleukins, thereby reducing inflammatory reactions. In addition, amino acids can also promote the production of antioxidants, such as glutathione, to remove free radicals in the body and protect cells from oxidative damage, thereby reducing inflammatory reactions. Therefore, proper amino acid supplementation is of great significance for maintaining physical health and preventing inflammatory diseases.

[0043] The role of amino acids in protecting cells: Amino acids can also protect cells from damage. When cells are damaged, amino acids can act as signal molecules to activate the protective mechanism within the cell, promote DNA repair and cell regeneration. In addition, amino acids can also promote cell repair and regeneration by providing energy and nutrients. Therefore, amino acids play an important role in maintaining cell health and repairing damaged tissues.

[0044] Sodium Hyaluronate:

[0045] Sodium hyaluronate acts as a tissue lubricant and is thought to play an important role in regulating the interactions between adjacent tissues. It forms a viscoelastic solution in water. The high viscosity of the solution provides mechanical protection to tissues (iris, retina) and cell layers (cornea, endothelium and epithelium). The elasticity of the solution helps absorb mechanical stress and provide protective cushioning to tissues. In promoting wound healing, it is thought to act as a protective transporter, bringing peptide growth factors and other structural proteins to the site of action. It is then enzymatically degraded and the active protein is released to promote tissue repair.

[0046] The role of each factor:

[0047] Vascular endothelial growth factor (VEGF):

[0048] VEGF can participate in restoring the tissue oxygen supply system. During the embryonic period, it regulates the proliferation, migration and survival of endothelial cells, thereby regulating the density and size of blood vessels, and then promoting the formation of new capillaries. In addition, the VEGF family also has a wide range of applications in the fields of inflammatory response, hematopoiesis, and neuroprotection.

[0049] Fibroblast growth factor (bFGF):

[0050] Basic fibroblast growth factor can promote cell growth, repair and replenish nutrition.

[0051] TNF-α (Tumor Necrosis Factor-α):

[0052] The mechanism of action of TNF-α (tumor necrosis factor-α) in vaginal relaxation is to affect the structure and function of vaginal tissue by regulating inflammatory response and cell apoptosis. TNF-α is an inflammatory mediator that can promote inflammatory response. In the vagina, the release of TNF-α can cause the infiltration and activation of inflammatory cells, leading to inflammatory symptoms such as tissue edema, redness, swelling, and pain.

[0053] Hepatocyte Growth Factor (HGF):

[0054] Hepatocyte growth factor (HGF) is a multifunctional factor that regulates the growth, motility and morphogenesis of various cells. Through paracrine or autocrine mechanisms, and with the help of epithelial-mesenchymal interactions, it plays an important role in embryogenesis, wound healing, angiogenesis, tissue and organ regeneration, morphogenesis and carcinogenesis.

[0055] Fibroblast Growth Factor FGF:

[0056] FGF, or fibroblast growth factor, is a multifunctional and powerful cytokine that plays an important role in promoting the metabolism of fibroblasts and the formation of collagen. Fibroblast growth factor can promote the growth and reproduction of skin tissue. It regulates the division, reproduction and growth differentiation of epithelial, endothelial and matrix cells of the skin by binding to specific receptors on the cell surface, promotes cell metabolism and enhances oxidation; it can promote the rapid growth and reproduction of cells related to skin damage, and regulate the synthesis, secretion and decomposition of intercellular matrix; it can promote the regeneration of stratum corneum cells, accelerate the repair of the stratum corneum and matrix layer of the skin, and promote the growth of human skin cells; it can enhance the synthesis of protein and cell metabolism of skin cells, and has the effect of delaying the aging of skin cells, promoting the repair and growth of epidermal cells, and making the skin smooth and plump.

[0057] The present invention has the following advantages:

[0058] The composition of the present invention can promote the rehabilitation treatment of cervicitis and cervical erosion with low risk and no need to worry about drug resistance. The composition of the present invention is highly multifunctional and has anti-inflammatory and immunomodulatory effects.

[0059] The composition of the present invention utilizes a multi-factor secretion extraction mixture to perform rehabilitation treatment on cervicitis and cervical erosion, that is, it utilizes factors to regulate and repair inflammation. It does not require customized treatment and can meet the needs of almost all patients. Even for patients with multiple underlying diseases, there is no need to worry about adverse reactions after use.

[0060] The cells of the composition of the invention have a high generation number, thus reducing the production cost. BRIEF DESCRIPTION OF THE DRAWINGS

[0061] In order to more clearly illustrate the implementation methods of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for the implementation methods or the description of the prior art. Obviously, the drawings in the following description are only exemplary, and for ordinary technicians in this field, other implementation drawings can be derived from the provided drawings without creative work.

[0062] The structures, proportions, sizes, etc. illustrated in this specification are only used to match the contents disclosed in the specification so as to facilitate understanding and reading by persons familiar with the technology. They are not used to limit the conditions under which the present invention can be implemented, and therefore have no substantial technical significance. Any structural modification, change in proportion or adjustment of size shall still fall within the scope of the technical contents disclosed in the present invention without affecting the effects and purposes that can be achieved by the present invention.

[0063] Figure 1 A comparison diagram of placental mesenchymal stem cells before and after induction provided by the present invention; wherein A-control group; B-induced group;

[0064] Figure 2 A comparison chart of cell viability counts of placental mesenchymal stem cells provided by the present invention; wherein A-control group; B-induced group;

[0065] Figure 3 The flow cytometric diagram provided by the present invention; wherein, A-control group; B-induced group;

[0066] Figure 4 A comparison chart of factors before and after induction of placental mesenchymal stem cells provided by the present invention;

[0067] Figure 5 A comparison chart of INF-γ in different groups of rats after treatment provided in Experimental Example 1 of the present invention;

[0068] Figure 6 A comparison chart of INF-γ in different groups of rats after treatment provided in Experimental Example 1 of the present invention;

[0069] Figure 7 The comparison chart before and after treatment provided for Case 1 of the present invention, wherein C-before treatment; D-after treatment;

[0070] Figure 8 The comparison chart before and after treatment provided for Case 2 of the present invention, wherein C-before treatment; D-after treatment;

[0071] Fig. 9 This is a comparison chart before and after treatment provided for Case 3 of the present invention, wherein C-before treatment; D-after treatment. DETAILED DESCRIPTION

[0072] The following is a description of the implementation of the present invention by specific embodiments. People familiar with the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0073] INF-γ (interferon-γ) is an important immunomodulatory factor that can promote the activation of immune cells such as macrophages, T lymphocytes and natural killer cells. INF-γ can promote the release of inflammatory mediators, such as proinflammatory cytokines and chemokines, guide the infiltration and activation of inflammatory cells, and aggravate the inflammatory response. In vaginitis, INF-γ affects the course of the disease by regulating immune cell activation, promoting inflammatory response, regulating cell apoptosis and affecting cell proliferation.

[0074] IL-1β can also promote cell proliferation and differentiation. In vaginitis, IL-1β may participate in the repair and regeneration of the vaginal mucosa by affecting the proliferation and differentiation of epithelial cells.

[0075] A cell supernatant extract after placental mesenchymal stem cell induction culture (after induction):

[0076] 1. Collecting rat placenta: Select SD rats weighing 200-250g. After the rats give birth to pups, quickly collect the placental tissue, remove the arteries and veins on the umbilical cord, cut the placenta into 1-1.5cm tissue blocks, rinse the umbilical cord with PBS, and finally stick the tissue blocks on a 75cm 2 Add 12 mL of serum-containing culture medium into the culture flask for culture.

[0077] 2. Cell culture: The isolated PMSCs were cultured at 37°C and 5% CO2. The culture medium was MEM (90%) + fetal bovine serum (10%) + penicillin (500 U / mL).

[0078] 3. Cultivate PMSCs: Change the medium after 3 days, and prevent the tissue blocks from floating when changing the medium. Change the medium once every 3 days, and generally remove the tissue blocks after 7-10 days (when the number of cells is small, the adhesion time can be appropriately extended). Continue to culture the cells until the fusion reaches about 85%, digest them with 0.25% trypsin, and pass them at a ratio of 1:3 (seeding density 1*10 5 / cm 2 ), and then passaged once every 3-5 days, and the fifth generation (F5) cells with good growth were selected for subsequent experiments.

[0079] 4. Identification and screening: The morphological characteristics observed under the microscope are spindle-shaped or stellate cells. Surface marker detection is performed. PMSCs positively express surface antigens: CD105, CD90, CD44 are strongly positive. Hematopoietic markers CD34, CD45 and HLA-DR are negatively expressed: they meet the surface marker characteristics of PMSCs and meet the identification standards of PMSCs.

[0080] 5. The fifth generation of placental mesenchymal stem cells were plated on a six-well plate and cultured in α-MEM medium. After the culture flask was placed in an incubator (5% CO2 / 37°C) for 72 hours, the growth density reached 80%, the cells grew well, the cell viability was >95%, and the morphology was consistent with the placental mesenchymal stem cell morphology. The results of cell surface molecular flow cytometry were consistent with the placental mesenchymal stem cell-related molecular indicators CD105, CD90, and CD44, which were strongly expressed, and the hematopoietic markers CD34.CD45 and HLA-DR were negatively expressed. The cell morphology observed under a microscope was consistent with the placental mesenchymal stem cell morphology. The supernatant was aspirated, and the residual liquid was aspirated and washed repeatedly with PBS buffer for 3 times. After washing, the residual liquid of PBS buffer was aspirated dry. After aspiration, 2 ml / well of induction medium (induction drugs: adenosine diphosphate heptose 5-15 μl / mL, protein kinase 3-10 ng / mL, vitamin E 5-20 μl / mL, dendrobium polysaccharide 5-15 μl / mL) was slowly added along the wall of the well plate. After the operation was completed, the well plate was placed in an incubator (5% CO2, 37°C) to induce bone marrow mesenchymal stem cells for 72 hours.

[0081] 6. Collect and purify the cytokines secreted by the cells (placental mesenchymal stem cell supernatant extract).

[0082] Control group (before induction): Placental stem cells were cultured in the same way without adding induction drugs.

[0083] Comparison of placental mesenchymal stem cells before and after induction Figure 1 As shown, after 72 hours of simultaneous culture in the control group and the induced group, the overall cell morphology was long spindle-shaped, with thinner cell ends, and there was no obvious change in the overall state of the cells, which was consistent with the morphology of placental mesenchymal stem cells.

[0084] Cell viability counts Figure 2 shown.

[0085] Flow cytometry images Figure 3 As shown in the figure, after 72h of cell culture, the induction group and the control group were digested and counted at the same time. The total cell concentration, live cell concentration and cell viability of the induction group were higher than those of the control group, and the proliferation ability of the cells in the induction group was stronger than that of the cells in the control group. The stemness of the control cells did not change before and after cell flow cytometry induction.

[0086] The results of ELISA test of factors before and after induction of placental mesenchymal stem cells are shown in Table 1, and the changes of inflammatory factors are shown in Figure 1. Figure 4 As shown, after induction, each factor increased significantly.

[0087] Table 1

[0088]

[0089] Example 1

[0090] This embodiment provides a composition for treating cervicitis and cervical erosion:

[0091] Placental mesenchymal stem cells PMSCs extract 4mL, vitamin C 10μl / mL, cyclosporine 5μg / mL, amino acid mixture 2mg / mL, sodium hyaluronate 0.01mg / ml.

[0092] Example 2

[0093] This embodiment provides a composition for treating cervicitis and cervical erosion:

[0094] Placental mesenchymal stem cells PMSCs extract 4.5mL, vitamin C 50μl / mL, cyclosporine 25μg / mL, amino acid mixture 15mg / mL, sodium hyaluronate 0.2mg / ml.

[0095] Example 3

[0096] This embodiment provides a composition for treating cervicitis and cervical erosion:

[0097] Placental mesenchymal stem cells PMSCs extract 5mL, vitamin C 100μl / mL, cyclosporine 45μg / mL, amino acid mixture 25mg / mL, sodium hyaluronate 0.5mg / ml.

[0098] Example 4

[0099] This embodiment provides a method for preparing a composition for treating cervicitis and cervical erosion:

[0100] Mix the raw materials evenly.

[0101] Experimental Example 1

[0102] 1. The following steps are required to create a rat vaginitis model:

[0103] 1. Prepare tools and materials: tweezers, gauze, syringe, sterile saline, 10% chloral hydrate, phenol glue, (Escherichia coli, Staphylococcus aureus, beta-hemolytic Streptococcus) mixed solution, iodine tincture, sterile gloves, and sterile mask.

[0104] 2. Select female rats weighing between 180-260g. Rearing conditions: temperature 18~22℃, relative humidity 40%~60%.

[0105] 3. Prepare (Escherichia coli, Staphylococcus aureus, and beta-hemolytic Streptococcus in a 2:1:1 ratio to make 2×10 9 / mL) mixture, use iodine to clean and disinfect the rat vulva and vagina, put the model SD rat head down, inject 0.1ml of mixed bacterial solution into the vagina 2cm deep, fix it with cotton balls and leave it in place for 2-3min to prevent the mixed solution from overflowing. After an interval of 12h, use the same method to infect once more, for a total of 6 times, and purulent discharge indicates that the model is successful.

[0106] 4. Identification of successful modeling: After infection, the rats' vaginal opening can be observed with the naked eye to see if there is redness, swelling, or purulent secretions. Use forceps to expand the vaginal opening of the rats to observe if the vaginal cervix is ​​enlarged, or there is water or pus. Observe the changes in the rats' weight before and after modeling.

[0107] Ten were reserved for modeling as the model group.

[0108] 2. Animal Drug Administration

[0109] After the modeling was completed, the modeling rats were randomly divided into 4 groups, 10 rats in each group, for a total of 40 rats, namely experimental group 1 (commercial drug), experimental group 2 (commercial drug), experimental group 3 (this composition) and blank group.

[0110] Dosage method: Use a sterile dosing device to administer the drug. Repeat the above steps again after 24 hours for 14 days.

[0111] Experimental group 1 (municipal drug): azithromycin (Suzhou Pharmaceutical Factory of Jiangsu Wuzhong Pharmaceutical Group Co., Ltd., national medicine standard H20010606, specification: 2.5 ml: 0.25 g) was dripped with a sterile applicator.

[0112] Experimental group 2 (municipal drug): Compound metronidazole cervical suppository (approval number: National Medicine Standard H13024553, production unit: Yaodu Pharmaceutical Group Co., Ltd., specification: 3g), placed deep in the vagina, once a day.

[0113] Experimental Group 3 (the composition of Example 2): The composition of Example 2 was injected deep into the vagina once a day.

[0114] Blank control group: injected with the same saline solution.

[0115] Model group: no treatment was given.

[0116] 3. Detection

[0117] After the last administration of the rats, they were fasted but not watered for 12 hours, and the rats in each group were anesthetized by intraperitoneal injection of 10% chloral hydrate solution. Blood was collected from the abdominal aorta into an EP tube, placed at room temperature for 2 hours, centrifuged at 3000r / min for 20 minutes, and the supernatant was collected to prepare serum, which was stored at -20°C. The levels of INF-γ (interferon-γ) and IL-1β in serum were determined using a microplate reader.

[0118] INF-γ Figure 5 and IL-1β Figure 6 As shown, after treatment, the drug of the present invention is slightly better than the commercially available drug.

[0119] The weight changes of rats before and after the experiment were statistically analyzed. As shown in Table 2, compared with the weight before the experiment, the weight of rats in the control group and the treatment group (experimental group 1, experimental group 2 and experimental group 3) was significantly lower than that in the model group. After treatment, the weight of experimental group 1, experimental group 2 and experimental group 3 was higher than that of the control group and close to that of the model group. After the experiment, the total number of white blood cells in the blood of rats in the treatment group was close to that of the model group, and the value of experimental group 3 was closer to the model group than that of experimental group 1 and experimental group 2, indicating a better treatment effect.

[0120] Table 2

[0121]

[0122] Experimental Example 2

[0123] Patients aged 30-50 years were selected, all of whom underwent cervical scraping to exclude cancer and secretion smear to exclude Trichomonas and Candida infection. 90 patients diagnosed with cervical erosion in the hospital were randomly divided into 3 groups, 30 in each group.

[0124] Dosage method: The patient should wash the vulva 3 days after the menstruation is over;

[0125] Experimental group 1 (commercial drug): azithromycin (Suzhou Pharmaceutical Factory of Jiangsu Wuzhong Pharmaceutical Group Co., Ltd., national medicine standard H20010606, specification: 2.5 ml: 0.25 g) was dripped with a sterile applicator.

[0126] Experimental group 2 (municipal drug): Compound metronidazole cervical suppository (approval number: National Medicine Standard H13024553, production unit: Yaodu Pharmaceutical Group Co., Ltd., specification: 3g), placed deep in the vagina, once a day.

[0127] Experimental Group 3 (composition of Example 2): injection deep into the vagina, once a day.

[0128] One course of treatment is 10 times, and the medicine is taken for 3 courses. Sexual intercourse is prohibited during the treatment.

[0129] Criteria for judging efficacy:

[0130] Significantly effective: the erosion area is reduced by >50% or the II degree erosion is converted to the I degree erosion, and the III degree erosion is converted to the II degree erosion;

[0131] Effective: The erosion area is reduced by < 50% or the erosion area is not significantly reduced, and the granular type is converted to the simple type, and the papillary type is converted to the granular type;

[0132] Ineffective: No change in the erosion area.

[0133] Statistical analysis: The data were processed using SPSS Winolons software, and the rank sum test was used as the statistical method, with P < 0.05 as the difference being statistically significant.

[0134] The results are shown in Table 3:

[0135] Table 3

[0136]

[0137] Case 1, Ms. Wang, 39 years old, cervical erosion stage II, was treated with intravaginal injection of the composition of Example 2 of the present invention. After 1 week of treatment, the effect was obvious; after 3 weeks of treatment, the patient was cured. Comparison of pictures before and after treatment Figure 7 shown.

[0138] Case 2, Ms. Zhang, 44 years old, cervical erosion stage II, was treated with intravaginal injection of the composition of Example 2 of the present invention. After 1 week of treatment, the effect was obvious; after 3 weeks of treatment, the patient was cured. Comparison of pictures before and after treatment Figure 8 shown.

[0139] Case 3, Ms. Li, 37 years old, cervical erosion stage III, was treated with intravaginal injection of the composition of Example 2 of the present invention. After 1 week of treatment, the effect was obvious; after 3 weeks of treatment, the patient was cured. Comparison of pictures before and after treatment Fig. 9 shown.

[0140] Although the present invention has been described in detail above by general description and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements can be made to the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all belong to the scope of protection claimed by the present invention.

Claims

1. A composition for treating gynecological diseases, characterized in that: The composition comprises 4-5 mL of placental mesenchymal stem cell PMSCs extract, 10-100 μl / mL of vitamin C, 5-45 μg / mL of cyclosporine, 2-25 mg / mL of amino acid mixture, and 0.01-0.5 mg / ml of sodium hyaluronate; The preparation method of the placental mesenchymal stem cell (PMSCs) extract comprises: culturing the placental mesenchymal stem cells using an α-MEM culture medium until the growth density is not less than 80%, the cells are in good growth state, and the cell viability is greater than 95%, removing the supernatant, adding an induction drug, and continuing the induction culture. After the culture is completed, the cytokines secreted by the cells are collected and purified to obtain the placental mesenchymal stem cell (PMSCs) extract; The induction drugs include 5-15 μL / mL of adenosine diphosphate heptose, 3-10 ng / mL of protein kinase, 5-20 μL / mL of vitamin E and 5-15 μL / mL of dendrobium polysaccharide.

2. A method for preparing a composition for treating gynecological diseases as claimed in claim 1, characterized in that: The method comprises mixing the raw materials.

3. Use of the composition for treating gynecological diseases as claimed in claim 1 in preparing a product for treating cervicitis.

4. Use of the composition for treating gynecological diseases as claimed in claim 1 in preparing a product for treating cervical erosion.

Images