Application of rare ginsenoside Rg5 in the preparation of drugs for inhibiting, improving or treating polycystic ovary syndrome

Abstract

The invention provides an application of rare ginsenoside Rg5 in preparing a drug for inhibiting, improving or treating polycystic ovary syndrome. Experiments have shown that rare ginsenoside Rg5 has a significant effect of promoting the proliferation of polycystic ovary syndrome-like ovarian granulosa cells and inhibiting, improving or treating polycystic ovary syndrome, and can be used to prepare a drug for inhibiting, improving or treating polycystic ovary syndrome and a drug for promoting the proliferation of polycystic ovary syndrome-like ovarian granulosa cells. The invention uses a polycystic ovary syndrome-like human ovarian granulosa cell model induced by oleic acid and androsol to demonstrate the application of rare ginsenoside Rg5 in promoting the proliferation of polycystic ovary syndrome-like ovarian granulosa cells, and uses a female SD rat model induced by letrozole and a high-sugar and high-fat feed to demonstrate the application of rare ginsenoside Rg5 in resisting polycystic ovary syndrome. The test method is safe and reliable, the test result is accurate, the therapeutic effect is significant, and there is no toxic side effect. The invention belongs to the field of medical technology.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to application of rare ginsenoside Rg5 in preparing a drug for inhibiting, improving or treating polycystic ovary syndrome. Background Art

[0002] Polycystic ovary syndrome (PCOS) is a common heterogeneous disease in women of childbearing age, with hyperandrogenemia, insulin resistance, hirsutism, acne, infertility and polycystic ovaries as the main clinical manifestations, and the incidence rate is 6% to 10%. The pathogenesis of PCOS is still unclear, and it may be closely related to factors such as inflammation, neuroendocrine, genetics and environment. High androgen is considered to be a key factor in impaired follicular development and metabolic disorders in PCOS, and high androgen increases the susceptibility to metabolic syndrome. PCOS patients have varying degrees of insulin resistance, and their risk of developing diseases such as type 2 diabetes or impaired glucose tolerance is higher than that of normal women. In addition, about 50% to 60% of PCOS patients are obese. Therefore, the treatment of PCOS has become a topic of common concern in the fields of gynecology and endocrinology.

[0003] At present, the drugs for treating PCOS are mainly chemical synthetic drugs, such as insulin sensitizers, ovulation drugs, and anti-androgen drugs. Chemical synthetic drugs have good efficacy in treating PCOS, but they have side effects, such as gastrointestinal irritation and difficulty in implantation of fertilized eggs. Therefore, developing an effective and safe plant compound to replace chemical synthetic drugs is an important development direction for the treatment of PCOS in the future.

[0004] Ginseng is a traditional Chinese medicine in my country, and its main active ingredient is ginsenoside. Ginsenoside has a variety of biological and pharmacological activities, such as anti-inflammatory, antioxidant, and anti-apoptotic, and has beneficial effects on cardiovascular diseases and metabolic syndrome. Rare ginsenosides are ginsenosides with lower polarity in ginseng. They have less sugar content, higher pharmacological activity, good permeability through cell membranes, and are more conducive to the body's absorption. They are more effective in anti-tumor, anti-diabetic, anti-thrombotic, regulating metabolism, and promoting cell differentiation. Among them, the rare ginsenoside Rg5 has strong pharmacological activities such as anti-tumor, lowering blood pressure, improving immunity, anti-inflammatory and improving lipid metabolism, but its effect on female reproduction has been less studied. Summary of the invention

[0005] In order to solve the technical problem that existing chemical synthetic drugs have side effects in treating polycystic ovary syndrome, the technical solution adopted in this application is to provide the use of rare ginsenoside Rg5 in the preparation of drugs for inhibiting, improving or treating polycystic ovary syndrome.

[0006] The embodiments of the present application provide the use of rare ginsenoside Rg5 in the preparation of a drug for inhibiting, improving or treating polycystic ovary syndrome.

[0007] The examples of the present application also provide the use of rare ginsenoside Rg5 in the preparation of a drug for promoting the proliferation of polycystic ovary syndrome-like ovarian granulosa cells.

[0008] In this application, the chemical structure of rare ginsenoside Rg5 (CAS: 186763-78-0) is:

[0009]

[0010] Preferably, the application of the rare ginsenoside Rg5 described above, calculated based on mammals other than humans, the dosage of the rare ginsenoside Rg5 used as a drug is 12.5-50 mg / kg / d.

[0011] In the present application, mammals other than humans may be rats, rabbits, sheep, and the like.

[0012] Preferably, the application of the rare ginsenoside Rg5 described above, calculated based on adults, the dosage of the rare ginsenoside Rg5 as a drug is 2.0-8.1 mg / kg / d.

[0013] In the present application, when rare ginsenoside Rg5 is used as a drug, its dosage varies depending on the degree of symptoms, patient age, gender, weight, sensitivity differences, administration method, administration period, administration interval, properties of the drug preparation, type of active ingredient, etc.

[0014] Preferably, the application of the rare ginsenoside Rg5 described above adopts a female SD rat model induced by letrozole and a high-sugar and high-fat diet to demonstrate the application of the rare ginsenoside Rg5 in treating polycystic ovary syndrome.

[0015] Preferably, the application of the rare ginsenoside Rg5 described above adopts a PCOS-like human ovarian granulosa cell model induced by oleic acid and androsandrone to demonstrate the application of the rare ginsenoside Rg5 in promoting the proliferation of PCOS-like ovarian granulosa cells.

[0016] Preferably, the application of the rare ginsenoside Rg5 described above, the use of the rare ginsenoside Rg5 as a medicine includes oral administration or external use.

[0017] In the present application, when the rare ginsenoside Rg5 is used as a medicine, its usage is not limited, and any suitable usage method can be used, depending on the specific circumstances; the dosage form of the rare ginsenoside Rg5 when used as a medicine is not limited. When it is used orally, the dosage form can be powder, capsule, granule, granule, tablet, pill, syrup, oral liquid; when it is used for external use, the dosage form can be a patch, smear, ointment.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] (1) The present invention provides the use of rare ginsenoside Rg5 in the preparation of drugs for inhibiting, improving or treating polycystic ovary syndrome. Experiments have found that rare ginsenoside Rg5 has a significant effect of promoting the proliferation of polycystic ovary syndrome-like ovarian granulosa cells and inhibiting, improving or treating polycystic ovary syndrome. It can be used to prepare drugs for inhibiting, improving or treating polycystic ovary syndrome and to prepare drugs for promoting the proliferation of polycystic ovary syndrome-like ovarian granulosa cells, which solves the technical problem of side effects of existing chemical synthetic drugs for the treatment of polycystic ovary syndrome and has good application prospects.

[0020] (2) The present invention uses a PCOS-like human ovarian granulosa cell model induced by oleic acid and androsol to demonstrate the application of rare ginsenoside Rg5 in promoting the proliferation of PCOS-like ovarian granulosa cells, and uses a female SD rat model induced by letrozole and a high-sugar and high-fat diet to demonstrate the application of rare ginsenoside Rg5 in treating PCOS. The test method is safe and reliable, the test results are accurate, the therapeutic effect is significant, and there are no toxic side effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0022] Figure 1 This is a graph showing the effects of oleic acid and androsol concentrations and intervention time on the survival rate of human ovarian granulosa cells in polycystic ovary syndrome;

[0023] Figure 2 The graphs are the changes in the survival rate of PCOS-like human ovarian granulosa cells in each group;

[0024] Figure 3 The graphs are the changes of testosterone content in human ovarian granulosa cells of PCOS in each group;

[0025] Figure 4These are the four stages of the estrus cycle of normal rats;

[0026] Figure 5 Figure 2 is the weight change of rats in each group;

[0027] Figure 6 It is the change diagram of fasting insulin (FINS) content of rats in each group;

[0028] Figure 7 The graphs are the changes of blood sugar in each group of rats;

[0029] Figure 8 The graphs are the changes in ovarian weight of rats in each group;

[0030] Fig. 9 The sections of ovarian tissues of rats in each group (light microscope, 100×);

[0031] Fig.10 The graphs are the changes of testosterone content in rats of each group;

[0032] Fig.11 The graph shows the changes in follicle stimulating hormone (FSH) levels in rats of each group. DETAILED DESCRIPTION

[0033] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present application more clearly understood, the present application is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present application and are not used to limit the present application.

[0034] The present invention provides the use of rare ginsenoside Rg5 in preparing a drug for inhibiting, improving or treating polycystic ovary syndrome. The raw materials and reagents used are conventional commercially available products, and their specific sources are shown in Table 1.

[0035] Table 1 Sources of raw materials and reagents

[0036]

[0037] In order to verify the reliability of the effect of the present invention, the present invention is described below in conjunction with Examples 1 to 3.

[0038] Example 1 Determination of the concentration and interference time of the modeling drug for polycystic ovary syndrome-like human ovarian granulosa cells

[0039] (1) Culture of human ovarian granulosa cells

[0040] Human ovarian granulosa cells were inoculated in a modified culture medium containing 10% fetal bovine serum and 100 U / mL penicillin and streptomycin, and cultured in a cell culture incubator at 37° C. and 5% CO 2 .

[0041] (2) Oleic acid and androsterone induction

[0042] 1×10 4 Human ovarian granulosa cells were seeded in 6-well plates and were divided into 6 groups after they attached to the wall. The experiment was repeated 3 times in each group. Then, equal volumes of oleic acid with concentrations of 0μM, 100μM, 200μM, 300μM, 400μM, and 500μM and equal volumes of andranolone with concentrations of 0nm, 1nm, 5nm, 10nm, 50nm, and 100nm were added to the 6 groups of well plates for intervention. The intervention was continued for 0h, 12h, 24h, and 48h to obtain human ovarian granulosa cells like polycystic ovary syndrome.

[0043] CCK-8 solution was added one hour before the end of the intervention. After the intervention, the absorbance of each well at 450nm was detected by using an enzyme marker, and the survival rate of PCOS-like human ovarian granulosa cells in each well was calculated. Then, a relationship diagram of the effects of oleic acid and androsol concentrations and intervention time on the survival rate of PCOS-like human ovarian granulosa cells was drawn, and the optimal oleic acid concentration, androsol concentration and intervention time were determined. The results are shown in Figure 2. Figure 1 shown.

[0044] Test conclusion:

[0045] See also Figure 1 , is a graph showing the effects of oleic acid and androsol concentrations and intervention time on the survival rate of human ovarian granulosa cells in polycystic ovary syndrome. Figure 1 It can be seen that the optimal concentration of oleic acid is 300 μM, the optimal concentration of androsol is 10 nm, and the cell survival rate is best after 48 hours of intervention, so the optimal intervention time is 48 hours.

[0046] Example 2 Test of the therapeutic effect of rare ginsenoside Rg5 on polycystic ovary syndrome-like human ovarian granulosa cells

[0047] In order to further confirm the reliability of the effect of the present invention, the following modeling conditions were selected: oleic acid concentration of 300 μM, andranolol concentration of 10 nm, and intervention time of 48 h, human ovarian granulosa cells were modeled, polycystic ovary syndrome-like human ovarian granulosa cells were obtained, and the therapeutic effect of rare ginsenoside Rg5 on polycystic ovary syndrome-like human ovarian granulosa cells was verified.

[0048] (1) Grouping

[0049] Six groups of experiments were set up to induce modeling of human ovarian granulosa cells to obtain PCOS-like human ovarian granulosa cells, and then the PCOS-like human ovarian granulosa cells were intervened by drugs. The specific induction drugs and intervention drugs of each group are shown in Table 2.

[0050] Table 2 Induction drugs and intervention drugs in each group

[0051]

[0052] (2) Culture of human ovarian granulosa cells

[0053] Human ovarian granulosa cells were inoculated in a modified culture medium containing 10% fetal bovine serum and 100 U / mL penicillin and streptomycin, and cultured in a cell culture incubator at 37° C. and 5% CO 2 .

[0054] (3) Oleic acid and androsanthellone induced modeling

[0055] 1×10 4 Human ovarian granulosa cells were seeded in 6-well plates and waited for adhesion. Then, 6 groups were divided, namely, con group, mod group, LGS group, MGS group, HGS group, and Met group. The experiment was repeated 3 times for each group. Then, equal volumes of oleic acid with a concentration of 300 μM and equal volumes of androsanthesine with a concentration of 10 nm were added to the well plates of mod group, LGS group, MGS group, HGS group, and Met group for intervention. The intervention was continued for 48 hours, and the modeling was completed. No treatment was given to the con group.

[0056] (4) Intervention with rare ginsenoside Rg5

[0057] After modeling, equal volumes of 2.5 mg / L rare ginsenoside Rg5, 5 mg / L rare ginsenoside Rg5, 10 mg / L rare ginsenoside Rg5, and 5 mg / L positive drug metformin hydrochloride were added to the well plates of the LGS group, MGS group, HGS group, and Met group for intervention for 48 hours; no drug intervention was performed in the mod group;

[0058] CCK-8 solution was added one hour before the end of the intervention. After the intervention, the absorbance of each well at 450 nm was detected using an enzyme marker, and the survival rate of PCOS-like human ovarian granulosa cells in each well was calculated. Then, the change graph of the survival rate of PCOS-like human ovarian granulosa cells in each group was drawn. The results are shown in Figure 2. Figure 2 shown.

[0059] (5) Hormone level testing

[0060] The supernatant of human ovarian granulosa cells in each well was obtained, and the supernatant of each well was treated with an Elisa kit, and then the absorbance of the supernatant of each well at 450nm was detected by an ELISA reader, and the testosterone content in the polycystic ovary syndrome-like human ovarian granulosa cells of each well was calculated according to the software provided by the Elisa kit, and the change graph of the testosterone content in the polycystic ovary syndrome-like human ovarian granulosa cells of each group was drawn. The results are shown in FIG. Figure 3 shown.

[0061] Test conclusion:

[0062] See also Figure 2 , is the change of survival rate of PCOS-like human ovarian granulosa cells in each group, where, compared with con, * indicates P<0.05, ** indicates P<0.01, *** indicates P<0.001, **** indicates P<0.0001; compared with mod group, # indicates P<0.05, ## indicates P<0.01, ### indicates P<0.001, #### indicates P<0.0001. Figure 2 It can be seen that, compared with the con group and the mod group, the survival rate of PCOS-like human ovarian granulosa cells in the mod group was significantly lower than that in the con group; compared with the mod group and the LGS group, the MGS group, and the HGS group, the survival rates of PCOS-like human ovarian granulosa cells in the LGS group, the MGS group, and the HGS group were significantly higher than those in the mod group, and the survival rate of PCOS-like human ovarian granulosa cells in the HGS group was the highest. It can be seen that the addition of rare ginsenoside Rg5 is beneficial to the proliferation of PCOS-like human ovarian granulosa cells, and the higher the concentration of rare ginsenoside Rg5, the better its effect on PCOS-like human ovarian granulosa cells. The better the proliferation effect of granulocytes, when the concentration of rare ginsenoside Rg5 is 10 mg / L, it has the best proliferation effect on PCOS-like human ovarian granulosa cells; comparing the Met group with the LGS group, MGS group, and HGS group, the survival rate of PCOS-like human ovarian granulosa cells in the LGS group was slightly lower than that in the Met group, while the survival rate of PCOS-like human ovarian granulosa cells in the MGS group and HGS group was significantly higher than that in the Met group. It can be seen that when the concentration of rare ginsenoside Rg5 is ≥5 mg / L, its proliferation effect on PCOS-like human ovarian granulosa cells is better than that of the positive drug metformin hydrochloride.

[0063] See also Figure 3 , is the change of testosterone content in PCOS-like human ovarian granulosa cells in each group, where, compared with con, * indicates P<0.05, ** indicates P<0.01, *** indicates P<0.001, **** indicates P<0.0001; compared with mod group, # indicates P<0.05, ## indicates P<0.01, ### indicates P<0.001, #### indicates P<0.0001. Figure 3It can be seen that, compared with the con group and the mod group, the testosterone content in the mod group was significantly higher than that in the con group, and the difference between the two was p<0.0001, and the difference was statistically significant. Therefore, oleic acid and androsol were successfully used for modeling; compared with the mod group and the LGS group, the MGS group, and the HGS group, the testosterone content in the LGS group, the MGS group, and the HGS group was significantly lower than that in the mod group, and the testosterone content in the HGS group was the lowest, and the difference between the two was p<0.001, and the difference was statistically significant. Therefore, after the intervention of the rare ginsenoside Rg5 in the HGS group, the testosterone content was significantly reduced, and the effect was the best; compared with the Met group and the LGS group, the MGS group, and the HGS group, the testosterone content in the LGS group was significantly higher than that in the Met group, the testosterone content in the MGS group was slightly higher than that in the Met group, and the testosterone content in the HGS group was significantly lower than that in the Met group. Therefore, when the concentration of the rare ginsenoside Rg5 was 10 mg / L, its effect on reducing the testosterone content was better than that of the positive drug metformin hydrochloride.

[0064] Example 3 In vivo therapeutic effect test of rare ginsenoside Rg5

[0065] (I) Modeling and intervention

[0066] (1) Animal grouping and modeling

[0067] Thirty-six SPF female SD rats of 6 weeks old were selected, with a mass of 160-180 g. The 36 rats were randomly divided into 6 groups, namely normal control group (con group), model control group (mod group), low-dose rare ginsenoside Rg5 group (LGS group), medium-dose rare ginsenoside Rg5 group (MGS group), high-dose rare ginsenoside Rg5 group (HGS group), and positive control group (Met group), with 6 rats in each group.

[0068] The rats in the con group were gavaged with 0.5% CMC-Na (10 mL / kg / d) in equal volume every day for 28 consecutive days. The rats in the other five groups were gavaged with letrozole (1 mg / kg / d) in equal volume every day and fed with high-sugar and high-fat diet in equal amounts for 28 consecutive days to induce PCOS rat model. The feeding conditions were room temperature 22±2℃, relative humidity 55±5%, and light-dark alternation conditions of 12h / 12h. The specific induction drugs and intervention drugs of each group are shown in Table 3.

[0069] Table 3 Inducing drugs and intervention drugs for rats in each group

[0070]

[0071] (2) Judgment of modeling standards

[0072] The criteria for successful modeling are: vaginal smears were taken from rats in each group at 10 a.m. every day from the 21st to the 28th day after induction with letrozole and high-sugar and high-fat diets, the cells smeared on the slides were fixed with methanol, the cells were stained with Giemsa staining solution, and then the main types of cells were observed under a microscope to determine the stage of the estrus cycle of the rats. The estrus cycle of normal rats is divided into the following 4 stages, as follows: Figure 4 As shown:

[0073] Figure 4 a is the estrus period: white blood cells are basically visible in the field of vision;

[0074] Figure 4 b is proestrus: a large number of nucleated epithelial cells can be seen, occasionally keratinized cells can be seen, and no leukocytes;

[0075] Figure 4 c is midestrus: almost all are anucleated keratinocytes, without leukocytes and nucleated epithelial cells;

[0076] Figure 4 d is late estrus: the number of cells is the largest in the field of view, and the amounts of the three types of cells are relatively uniform.

[0077] The estrous cycle of normal rats shows regular changes between proestrus, metaestrus, metestrus and diestrus. When the estrous cycle of rats in the mod group is disordered and irregular, it means that the model is successfully established and PCOS rats are obtained.

[0078] (3) Intervention with rare ginsenoside Rg5

[0079] After the modeling, rats in the con group and mod group were given distilled water gavage once a day; rats in the LGS group, MGS group, and HGS group were given 12.5 mg / kg / d, 25 mg / kg / d, and 50 mg / kg / d of rare ginsenoside Rg5 gavage once a day; rats in the Met group were given 180 mg / kg / d of metformin hydrochloride gavage once a day; and the equal volume gavage was continued for 21 days. At the beginning of gavage treatment, the body weight of rats in each group was measured on 0d, 2d, 4d, 6d, 8d, 10d, 12d, 14d, 16d, 18d, and 20d, and the weight change graph of rats in each group was drawn. The results are shown in Figure 2. Figure 5 As shown; during the oral gavage treatment, the rats were maintained with free access to food and clean drinking water.

[0080] Test conclusion:

[0081] See also Figure 5 , is the weight change diagram of rats in each group. Figure 5It can be seen that the body weights of rats in the con group and the mod group showed an increasing trend, and the body weight of rats in the mod group was always higher than that of the other five groups; the body weights of rats in the LGS group, MGS group, HGS group and Met group showed a decreasing trend, and the body weights of rats in the LGS group, MGS group and HGS group were close to that of rats in the Met group. It can be seen that the rare ginsenoside Rg5 can reduce the body weight of PCOS rats, and its dosage is lower than that of metformin hydrochloride.

[0082] (II) Detection of the improvement effect of rare ginsenoside Rg5 on PCOS rats

[0083] (1) Fasting blood glucose (FBG), fasting insulin (FINS) levels and glucose tolerance test

[0084] One day before the end of the intragastric administration, the rats in each group were fasted but not watered for 12 h, and then the tails were cut to collect blood. The fasting blood glucose (FBG) of the rats in each group was measured using a blood glucose meter, and the fasting insulin (FINS) content of the rats in each group was measured by enzyme-linked immunosorbent assay using an Elisa kit. The changes in the fasting insulin (FINS) content of the rats in each group were plotted. The results are shown in Figure 2. Figure 6 As shown;

[0085] After the fasting blood glucose (FBG) was measured, each group of rats was orally gavaged with 50wt% glucose solution at a dose of 2g / kg. The blood glucose of each group of rats was then monitored at 30min, 60min, 90min, and 120min after gavage, and the changes in blood glucose of each group of rats were plotted. The results are shown in Figure 2. Figure 7 shown.

[0086] Test conclusion:

[0087] See also Figure 6 , is the change diagram of fasting insulin (FINS) content of rats in each group, among which, compared with con group, * indicates P<0.05, ** indicates P<0.01, *** indicates P<0.001, **** indicates P<0.0001; compared with mod group, # indicates P<0.05, ## indicates P<0.01, ### indicates P<0.001, #### indicates P<0.0001. Please refer to Figure 7 , is the change diagram of blood glucose in each group of rats, where the blood glucose at 0min is fasting blood glucose (FBG). Figure 6 and Figure 7It can be seen that, compared with the con group and the mod group, the FBG content of the rats in the mod group was significantly increased, and the FINS content of the rats in the mod group was significantly increased, p<0.05, and the difference was statistically significant. It can be seen that the rats in the mod group had insulin resistance, that is, after letrozole and high-sugar and high-fat diet modeling, the rats had insulin resistance; comparing the mod group and the LGS group, MGS group, and HGS group, the FBG content of the rats in the LGS group, MGS group, and HGS group was significantly decreased, and the FINS of the rats in the HGS group was significantly decreased, p<0.05, and the difference was statistically significant. It can be seen that the rare ginsenoside Rg5 in the HGS group can improve the insulin resistance of PCOS rats.

[0088] See also Figure 7 , is the change diagram of blood sugar in each group of rats. Figure 7 It can be seen that the glucose clearance of rats in the mod group was delayed and the area under the curve increased, which shows that oral administration of letrozole and feeding a high-sugar and high-fat diet reduced the glucose excretion ability of rats; after administration of rare ginsenoside Rg5, the area under the curve decreased, which shows that rare ginsenoside Rg5 enhanced the glucose excretion ability of rats and improved the impairment of glucose tolerance; comparing the mod group with the LGS group, MGS group, and HGS group, the area under the curve of the HGS group was the smallest and the improvement effect was the best.

[0089] (2) Ovarian quality and ovarian morphology detection

[0090] After 21 days of drug treatment, rats in each group were fasted for 12 hours and anesthetized by inhalation of isoflurane. After successful anesthesia, they were locally disinfected with medical ethanol, the abdominal cavity was opened, and 5 mL of blood was collected through the abdominal aorta. The blood was centrifuged at 4000 r / min for 15 minutes, and the supernatant was divided into centrifuge tubes and then frozen in a -80℃ refrigerator for later use.

[0091] The ovaries of the rats were separated and washed with saline and dried with absorbent paper. The ovaries were weighed and the changes in the ovarian weight of the rats in each group were plotted. The results are shown in Figure 2. Figure 8 As shown;

[0092] The ovaries of rats in each group were fixed in 4% paraformaldehyde solution and sent to Wuhan Sewell Biotechnology Co., Ltd. after 24 hours. After paraffin embedding and hematoxylin-eosin (HE) staining, the sections were observed under an inverted microscope. The sections of the ovarian tissues of rats in each group are shown in the following figure. Fig. 9 shown.

[0093] Test conclusion:

[0094] See also Figure 8, is the change diagram of ovarian weight of rats in each group, where, compared with con group, * indicates P<0.05, ** indicates P<0.01, *** indicates P<0.001, **** indicates P<0.0001; compared with mod group, # indicates P<0.05, ## indicates P<0.01, ### indicates P<0.001, #### indicates P<0.0001. Figure 8 It can be seen that 21 days after administration, the ovarian mass of rats in the mod group was significantly increased compared with the con group and the mod group, and the difference was statistically significant (P<0.001); the ovarian mass of rats in the mod group was significantly decreased compared with the LGS group, the MGS group, and the HGS group, and the difference was statistically significant (P<0.05). It can be seen that the three doses of rare ginsenoside Rg5 can reduce the increase in ovarian mass of PCOS rats caused by letrozole combined with high-sugar and high-fat feed, and promote the recovery of ovarian mass of PCOS rats to normal levels.

[0095] See also Fig. 9 , are the sections of ovarian tissues of rats in each group. The presence of multiple dilated cystic follicles in the ovaries is a typical feature of PCOS. After HE staining, the ovarian specimens of rats in each group were observed under a microscope. Compared with the con group and the mod group, there were multiple corpora lutea in the ovaries of rats in the con group, which were neatly arranged and complete in morphology, with very few cystic dilated follicles. The corpora lutea in the ovaries of rats in the mod group were smaller in size and fewer in number, with more cystic dilated follicles and atretic follicles, and the granulosa layer was disordered and the number of layers was reduced. Compared with the mod group and the LGS group, the MGS group, and the HGS group, the number of corpora lutea in the ovaries of rats in the MGS group and the HGS group increased, and cystic dilated follicles were rare. It can be seen that the rare ginsenoside Rg5 in the MGS group and the HGS group can effectively improve the ovarian lesions of rats with PCOS.

[0096] (3) Sex hormone level testing

[0097] The serum supernatants of the six groups of rats were taken out and treated with Elisa kits. Then, the absorbance of the serum supernatants of each group at 450 nm was detected with an ELISA reader. The contents of follicle stimulating hormone (FSH) and testosterone (T) in the rats were calculated according to the software provided by the Elisa kits. Then, the change graphs of testosterone content and follicle stimulating hormone (FSH) content in the rats of each group were drawn. The results were as follows: Fig.10 , Fig.11 shown.

[0098] Test conclusion:

[0099] See also Fig.10, is the change diagram of testosterone content in rats of each group, among which, compared with con group, * indicates P<0.05, ** indicates P<0.01, *** indicates P<0.001, **** indicates P<0.0001; compared with mod group, # indicates P<0.05, ## indicates P<0.01, ### indicates P<0.001, #### indicates P<0.0001. Fig.10 It can be seen that, compared with the con group and the mod group, the testosterone content in the mod group was significantly increased, and the difference was statistically significant (P<0.001); compared with the mod group and the LGS group, the MGS group, and the HGS group, the testosterone content in the LGS group was significantly decreased, and the difference was not statistically significant, and it was dose-dependent. The testosterone content in the MGS group was significantly decreased, and the difference was statistically significant (P<0.01), and the testosterone content in the HGS group was significantly decreased, and the difference was statistically significant (P<0.01). It can be seen that the rare ginsenoside Rg5 in the MGS group and the HGS group can effectively improve the increased testosterone content in the serum of PCOS rats.

[0100] See also Fig.11 , is the change diagram of follicle stimulating hormone (FSH) content in each group of rats, among which, compared with con group, * indicates P<0.05, ** indicates P<0.01, *** indicates P<0.001, **** indicates P<0.0001; compared with mod group, # indicates P<0.05, ## indicates P<0.01, ### indicates P<0.001, #### indicates P<0.0001. Fig.11 It can be seen that, compared with the con group and the mod group, the FSH content in the mod group was significantly decreased, and the difference was statistically significant (P<0.05); compared with the mod group and the LGS group, the MGS group and the HGS group, the FSH content in the LGS group and the MGS group was significantly increased, and the difference was not statistically significant, and the FSH content in the HGS group was also significantly increased, and the difference was statistically significant (P<0.05). It can be seen that the rare ginsenoside Rg5 in the HGS group can increase the production of follicle-stimulating hormone in the serum of PCOS rats.

[0101] Comprehensive analysis Figure 5-Figure 11 The rare ginsenoside Rg5 has the effect of promoting the proliferation of PCOS-like ovarian granulosa cells in PCOS rats induced by letrozole and high-sugar and high-fat diet, and inhibiting or improving or treating PCOS.

[0102] The present invention also provides the use of rare ginsenoside Rg5 in preparing a drug for inhibiting, improving or treating polycystic ovary syndrome, and the use of rare ginsenoside Rg5 in preparing a drug for promoting the proliferation of polycystic ovary syndrome-like ovarian granulosa cells; when the rare ginsenoside Rg5 is used as a drug, the dosage thereof is 12.5 to 50 mg / kg / d calculated for mammals other than humans, and the dosage thereof is 2.0 to 8.1 mg / kg / d calculated for adults; the use of the rare ginsenoside Rg5 as a drug includes oral administration or external use.

[0103] In summary, the present invention provides the use of rare ginsenoside Rg5 in preparing drugs for inhibiting, improving or treating polycystic ovary syndrome. Experiments have found that rare ginsenoside Rg5 has a significant effect of promoting the proliferation of polycystic ovary syndrome-like ovarian granulosa cells and inhibiting, improving or treating polycystic ovary syndrome. It can be used to prepare drugs for inhibiting, improving or treating polycystic ovary syndrome and to prepare drugs for promoting the proliferation of polycystic ovary syndrome-like ovarian granulosa cells, which solves the technical problem that existing chemical synthetic drugs have side effects in treating polycystic ovary syndrome and has good application prospects. The present invention uses a polycystic ovary syndrome-like human ovarian granulosa cell model induced by oleic acid and androsan to prove the use of rare ginsenoside Rg5 in promoting the proliferation of polycystic ovary syndrome-like ovarian granulosa cells, and uses a female SD rat model induced by letrozole and high-sugar and high-fat feed to prove the use of rare ginsenoside Rg5 in anti-polycystic ovary syndrome. The test method is safe and reliable, the test results are accurate, the therapeutic effect is significant, and there are no toxic side effects; it can be widely used in the field of medical technology.

[0104] The embodiments described above are only used to illustrate the technical solutions of the present application, rather than to limit them. Although the present application has been described in detail with reference to the aforementioned embodiments, a person skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. Such modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present application, and should all be included in the protection scope of the present application.

Claims

1. The use of rare ginsenoside Rg5 as the sole active ingredient in the preparation of drugs for inhibiting, improving or treating polycystic ovary syndrome.

2. The use of the rare ginsenoside Rg5 according to claim 1, characterized in that: Calculated based on mammals other than humans, the dosage of rare ginsenoside Rg5 used as a drug is 12.5-50 mg / kg / d.

3. The use of the rare ginsenoside Rg5 according to claim 1, characterized in that: Calculated based on adults, the dosage of rare ginsenoside Rg5 used as a drug is 2.0-8.1 mg / kg / d.

4. The use of the rare ginsenoside Rg5 according to claim 1, characterized in that: A female SD rat model induced by letrozole and high-sugar and high-fat diet was used to demonstrate the application of rare ginsenoside Rg5 in anti-polycystic ovary syndrome.

5. The use of the rare ginsenoside Rg5 according to claim 1, characterized in that: The rare ginsenoside Rg5 is used as a drug by oral administration.

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