Use of sanqi danshen tablet in treatment of polycystic ovary syndrome

Abstract

The present application provides the use of Sanqi Danshen tablets in the treatment of polycystic ovary syndrome, belonging to the technical field of traditional Chinese medicine. The present application provides the use of Sanqi Danshen tablets in the preparation of drugs, especially in the preparation of drugs for treating polycystic ovary syndrome. The animal model of the present application proves that Sanqi Danshen tablets can reduce the levels of testosterone (T), luteinizing hormone (LH), anti-Mullerian hormone (AMH) and insulin resistance index (HOMA-IR), thereby improving polycystic ovarian lesions. The drug described in the present application has the effects of regulating hormone levels, relieving insulin resistance, inhibiting cystic dilatation of follicles, allowing normal development of follicles, and ultimately improving the structure and function of the ovary.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology, specifically relating to the use of Sanqi Danshen tablets in the treatment of polycystic ovary syndrome. Background Technology

[0002] Polycystic ovary syndrome (PCOS) is a common multisystem disorder characterized by reproductive endocrine disorders such as hyperandrogenemia (HA), menstrual irregularities, polycystic ovarian morphology, and infertility. It is often accompanied by metabolic disorders, cardiovascular disease, sleep apnea, mood disorders, and even tumors. PCOS not only affects female fertility but is also closely related to various metabolic diseases, including diabetes and cardiovascular disease, seriously threatening women's health. In Traditional Chinese Medicine (TCM), PCOS falls under the categories of "menstrual disorders," "amenorrhea," and "abdominal masses," with its pathogenesis often related to kidney deficiency, liver stagnation, spleen deficiency, phlegm-dampness, and blood stasis.

[0003] Sanqi Danshen tablets use Danshen as the main ingredient, supplemented with Sanqi and Xiangfu, which precisely cover the core pathogenesis of PCOS such as kidney deficiency, spleen deficiency, liver stagnation and blood stasis, and show significant therapeutic effects in rat PCOS models. Summary of the Invention

[0004] This invention provides the use of Panax notoginseng and Salvia miltiorrhiza tablets in the treatment of polycystic ovary syndrome (PCOS), and the Panax notoginseng and Salvia miltiorrhiza tablets have a good therapeutic effect on PCOS model rats.

[0005] This invention provides the application of Panax notoginseng and Salvia miltiorrhiza tablets in the preparation of medicines, wherein the medicines have at least one of the following effects: regulating hormone levels, relieving insulin resistance, inhibiting cystic dilated follicles, promoting normal follicle development, and improving ovarian tissue structure.

[0006] In a preferred embodiment of the present invention, the regulation of hormone levels includes reducing the level of at least one of the following hormones: testosterone, luteinizing hormone, anti-Müllerian hormone, and follicle-stimulating hormone.

[0007] This invention also provides the application of Panax notoginseng and Salvia miltiorrhiza tablets in the preparation of drugs for treating polycystic ovary syndrome.

[0008] In a preferred embodiment of the present invention, the working concentration of Panax notoginseng and Salvia miltiorrhiza tablets in the drug is greater than 2.5 g / kg.

[0009] Beneficial Effects: This invention established a rat model of polycystic ovary syndrome (PCOS) using a combination of insulin and human chorionic gonadotropin (hCG). The model was then treated with various drugs, and serum levels of testosterone (T), luteinizing hormone (LH), anti-Müllerian hormone (AMH), follicle-stimulating hormone (FSH), and fasting insulin were measured. Ovaries were also harvested for pathological examination. Compared to the control group, rats in each treatment group showed significantly lower levels of testosterone (T), luteinizing hormone (LH), and the homeostasis model of insulin resistance (HOMA-IR), and significant improvement in polycystic ovarian symptoms. Sanqi Danshen tablets can improve ovarian lesions and insulin resistance by reducing T, LH, AMH, LH / FSH, and HOMA-IR levels. High-dose Sanqi Danshen suspension (5 g / kg) was superior to low-dose Sanqi Danshen suspension (2.50 g / kg) in reducing LH / FSH, AMH, and the number of cystic dilated follicles. Its therapeutic effect in reducing AMH and LH / FSH was superior to that of the commercially available drug metformin. The above results indicate that Panax notoginseng and Salvia miltiorrhiza tablets can regulate hormone levels, alleviate insulin resistance, thereby inhibiting cystic dilated follicles, promoting normal follicle development, and ultimately improving ovarian tissue structure. Furthermore, high-dose Panax notoginseng and Salvia miltiorrhiza suspension has a better therapeutic effect. Attached Figure Description

[0010] Figure 1 The graph shows the changes in rat body weight during the drug administration process. In the graph, "Con" represents the normal control group, "PCOS" represents the model control group, "SQDS-L" represents the low-dose group of Panax notoginseng and Salvia miltiorrhiza tablets (250 mg / kg), "SQDS-H" represents the high-dose group of Panax notoginseng and Salvia miltiorrhiza tablets (500 mg / kg), and "Met" represents the metformin group (200 mg / kg). Figure 2 The figure shows the effect of Panax notoginseng and Salvia miltiorrhiza tablets on hormone levels in PCOS rats. In the figure, "Con" represents the normal control group, "PCOS" represents the model control group, "SQDS-L" represents the low-dose group of Panax notoginseng and Salvia miltiorrhiza tablets (250 mg / kg), "SQDS-H" represents the high-dose group of Panax notoginseng and Salvia miltiorrhiza tablets (500 mg / kg), and "Met" represents the metformin group (200 mg / kg). Compared with the normal control group, P<0.05, P<0.01; # represents the difference between the model control group and the control group, #P<0.05, ##P<0.01; AE: T, AMH, FSH, LH and LH / FSH levels of rats in each group at the end of drug administration; Figure 3The figure shows the effect of Panax notoginseng and Salvia miltiorrhiza tablets on insulin resistance in PCOS rats. In the figure, "Con" represents the normal control group, "PCOS" represents the model control group, "SQDS-L" represents the low-dose group of Panax notoginseng and Salvia miltiorrhiza tablets (250 mg / kg), "SQDS-H" represents the high-dose group of Panax notoginseng and Salvia miltiorrhiza tablets (500 mg / kg), and "Met" represents the metformin group (200 mg / kg). Compared with the normal control group, P<0.05, P<0.01; # represents the difference between the model control group and the control group, #P<0.05, ##P<0.01; A: Fasting glucose level of rats in each group after drug administration; B: Fasting insulin level of rats in each group after drug administration; C: Insulin resistance index (HOMA-IR) of rats in each group after drug administration. Figure 4 The figure shows the effects of Panax notoginseng and Salvia miltiorrhiza tablets on the uterine and ovarian coefficients of PCOS rats. In the figure, "Con" represents the normal control group, "PCOS" represents the model control group, "SQDS-L" represents the low-dose Panax notoginseng and Salvia miltiorrhiza group (250 mg / kg), "SQDS-H" represents the high-dose Panax notoginseng and Salvia miltiorrhiza group (500 mg / kg), and "Met" represents the metformin group (200 mg / kg). A: Uterine coefficient of rats in each group at the end of drug administration; B: Ovarian coefficient of rats in each group at the end of drug administration. Figure 5 The figure shows the effect of Panax notoginseng and Salvia miltiorrhiza tablets on the morphology of polycystic ovaries in PCOS rats. In the figure, "Con" represents the normal control group, "PCOS" represents the model control group, "SQDS-L" represents the low-dose group of Panax notoginseng and Salvia miltiorrhiza tablets (250 mg / kg), "SQDS-H" represents the high-dose group of Panax notoginseng and Salvia miltiorrhiza tablets (500 mg / kg), and "Met" represents the metformin group (200 mg / kg). A: Typical pathological images of ovaries in each group after drug administration, with arrows indicating cystic dilated follicles; B: Number of polycystic ovaries in each group after drug administration. Detailed Implementation

[0011] This invention provides the application of Panax notoginseng and Salvia miltiorrhiza tablets in the preparation of medicines, wherein the medicines have at least one of the following effects: regulating hormone levels, relieving insulin resistance, inhibiting cystic dilated follicles, promoting normal follicle development, and improving ovarian tissue structure.

[0012] In this embodiment of the invention, a rat PCOS model was constructed and the model was treated with medication. The results showed that after administration of Sanqi Danshen tablets, the levels of testosterone (T), luteinizing hormone (LH), anti-Müllerian hormone (AMH), and insulin resistance index (HOMA-IR) in rats were significantly reduced, and the polycystic ovarian symptoms were significantly improved. This proves that Sanqi Danshen tablets may improve hormone levels in PCOS rats and inhibit polycystic ovarian changes by reducing T, LH, AMH, and LH / FSH levels; and improve insulin resistance by reducing HOMA-IR levels.

[0013] This invention does not specifically limit the source of the Panax notoginseng and Salvia miltiorrhiza tablets, requiring that Salvia miltiorrhiza be the main ingredient, with other components including Panax notoginseng, Cyperus rotundus, etc. In the examples, the Panax notoginseng and Salvia miltiorrhiza tablets used were developed by the First Affiliated Hospital of Kunming Medical University and have been patented; its Chinese patent application number is CN1151887A. In this invention, the Panax notoginseng and Salvia miltiorrhiza tablets consist of Salvia miltiorrhiza or its extract, Panax notoginseng, and Cyperus rotundus, with Salvia miltiorrhiza or its extract as the main ingredient, containing 55-65%. Furthermore, the Salvia miltiorrhiza extract can be obtained by directly grinding Salvia miltiorrhiza into powder or by extraction with water or organic solvents to obtain a precipitate, which is the Salvia miltiorrhiza extract described in this invention.

[0014] This invention provides the application of Panax notoginseng and Salvia miltiorrhiza tablets in the preparation of drugs for treating polycystic ovary syndrome.

[0015] The dosage form of the drug described in this invention can be a suspension, granules, tablets, or capsules, etc. In one embodiment of this invention, a suspension is used as an example, and 0.5% CMC-Na solution is used as a solvent when preparing the suspension.

[0016] In a preferred embodiment of the present invention, the working dose of Panax notoginseng and Salvia miltiorrhiza tablets in the pharmaceutical composition is greater than 2.50 g / kg.

[0017] To further illustrate the present invention, the use of the Panax notoginseng and Salvia miltiorrhiza tablets provided by the present invention in the treatment of polycystic ovary syndrome is described in detail below with reference to embodiments, but these should not be construed as limiting the scope of protection of the present invention.

[0018] Unless otherwise specified, all materials used in the embodiments of the present invention are conventional commercially available materials, and the test methods used are also conventional methods in the field.

[0019] The SPF-grade female SD rats used in this embodiment of the invention totaled 54, weighing 180-200 g, and were purchased from the Henan Provincial Animal Experiment Center. The animal quality certificate number is: No. 411006251100013236. The rats were housed in the SPF animal facility of the Institute of Materia Medica, Zhengzhou University. The facility license number is SYXK(Yu)2023 - 0004. During the experiment, the rats had free access to food and water, and the room temperature was maintained at 20 - 22 °C with a relative humidity of 60% - 70%.

[0020] The main reagents used in the embodiments of the present invention and their sources are shown in Table 1 below.

[0021] Table 1 Main reagents in the embodiments of the present invention

[0022] The preparation methods of the main reagents involved in the embodiments of the present invention are as follows:[[]]END]] 1. 0.5% CMC - Na solution: Measure 400 mL of distilled water with a measuring cylinder and place it in a conical flask. Use an electronic balance to accurately weigh 2.0000 g of CMC - Na powder. Slowly add the CMC - Na powder to the distilled water while stirring until the powder is completely dissolved.

[0023] 2. Panax notoginseng and Salvia miltiorrhiza tablet suspension: Take 5 Panax notoginseng and Salvia miltiorrhiza tablets (specification: 0.26 g / tablet), grind them into fine powder in a mortar, add 0.5% CMC - Na solution, continue to grind finely and then transfer to a 50 mL EP tube, dilute to 50 mL with 0.5% CMC - Na solution, and perform low - temperature ultrasonic treatment for 20 min to obtain a low - dose Panax notoginseng and Salvia miltiorrhiza tablet suspension (25 mg / mL). Take another 10 Panax notoginseng and Salvia miltiorrhiza tablets and prepare to 50 mL in the same way to obtain a high - dose Panax notoginseng and Salvia miltiorrhiza tablet suspension (50 mg / mL).

[0024] 3. Metformin suspension (20 mg / mL): Take 2 metformin hydrochloride tablets (specification: 0.5 g / tablet), grind them into fine powder in a mortar, add 0.5% CMC - Na solution, continue to grind finely and then transfer to a 50 mL EP tube, and perform low - temperature ultrasonic treatment for 20 min.

[0025] The experimental data obtained in the embodiments of the present invention were processed using SPSS 27.0 statistical software, and the results were expressed as mean ± standard deviation ( ); for comparing the means of two samples, t - test was used, and for comparing the means of multiple samples, one - way analysis of variance (ANVOA) was used. The results were considered significant at P = 0.05 and extremely significant at P = 0.01.

[0026] Example 1 1. Modeling method and drug administration Female SD rats were randomly divided into two groups: a normal control group (n=9) and a model group (n=45). From day 1 to 10, the model group received subcutaneous insulin injections, starting at 0.5 IU / day and increasing by 0.5 IU daily to 4.0 IU (day 8). On days 9 and 10, the insulin was increased by 1.0 IU daily, gradually increasing to 6.0 IU / day, and maintained at this dose until day 22. Simultaneously, from day 11 to 22, each animal received a subcutaneous injection of 3.0 IU of HCG solution once daily, with 5% glucose solution replacing their daily drinking water. The normal control group received the same volume of physiological saline. On the last day of modeling, the rats were fasted overnight, and blood was collected from the jugular vein. A significant increase in serum testosterone levels was considered a successful model. Ovaries from two additional animals were harvested for pathological analysis; the presence of polycystic ovaries was considered a successful model.

[0027] After successful modeling, the PCOS model rats were randomly divided into 4 groups: Model control group: Administered 0.5% CMC-Na at a dose of 10 mL / kg, once daily for a total of 28 days; Low-dose group of Panax notoginseng and Salvia miltiorrhiza tablets: 25 mg / mL Panax notoginseng and Salvia miltiorrhiza tablet suspension, the dosage is 10 mL / kg, once a day, for a total of 28 days; High-dose group of Panax notoginseng and Salvia miltiorrhiza tablets: 50 mg / mL Panax notoginseng and Salvia miltiorrhiza tablet suspension, dosage 10 mL / kg, once a day, for a total of 28 days; Metformin group: 20 mg / mL metformin suspension, administered at a dose of 10 mL / kg, once daily for a total of 28 days.

[0028] 2. Detection indicators 2.1 Body weight: During the administration period, the animal's body weight was measured every 4 days.

[0029] During the modeling period in female SD rats, compared with the normal control group, the water intake and urine output increased; during the drug administration period, compared with the normal control group, no obvious abnormal signs were observed in any of the drug administration groups.

[0030] The changes in body weight of rats in each group are as follows: Figure 1 As shown, during the administration process, there was no significant change in the body weight of rats in each group, and no statistically significant difference was observed between the groups.

[0031] 2.2 Hormone Levels: One day prior to the end of administration, the animals were fasted, anesthetized with isoflurane, and their abdomens were opened. Blood was collected from the abdominal aorta, and serum was separated to detect the levels of testosterone (T), luteinizing hormone (LH), anti-Müllerian hormone (AMH), and fasting insulin (FINS).

[0032] Hormone level detection results of rats in each group 28 days after drug administration are as follows: Figure 2 As shown, compared with the normal control group, the levels of T, AMH, LH, and LH / FSH in the model control group rats were significantly increased, while FSH was significantly decreased. Compared with the model control group, the T level in the high- and low-dose Sanqi Danshen tablet groups and the metformin group rats was significantly decreased (P<0.01); the AMH level in the high-dose Sanqi Danshen tablet group rats was significantly decreased (P<0.05), while the AMH level in the metformin group rats showed no significant change. Compared with the model control group, the FSH level in the metformin group rats was significantly decreased (P<0.05); the LH level in the high- and low-dose Sanqi Danshen tablet groups and the metformin group rats was significantly decreased (P<0.05); the serum LH / FSH ratio in the low- and high-dose Sanqi Danshen tablet groups rats was significantly decreased, and this decrease was dose-dependent.

[0033] 2.3 Insulin resistance: Fasting plasma glucose (FPG) levels were measured and recorded using a blood glucose meter and blood glucose test strips, and the Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) index was calculated (HOMA-IR = FPG × FINS / 22.5).

[0034] Insulin resistance in rats of each group 28 days after administration is as follows: Figure 3 As shown, there were no significant differences in fasting blood glucose levels among the groups after drug administration. Compared with the normal control group, the INS level of rats in the model control group was significantly increased (P<0.01); compared with the model control group, the INS level of rats in the metformin group was significantly decreased (P<0.05), and there were no significant changes in INS levels in the low-dose and high-dose groups of Sanqi Danshen tablets.

[0035] Compared with the normal control group, the HOMA-IR of rats in the model control group was significantly increased (P<0.01); compared with the model control group, the HOMA-IR of rats in the low-dose and high-dose groups of Sanqi Danshen tablets was significantly decreased (P<0.05), and the HOMA-IR of rats in the metformin group was significantly decreased (P<0.01).

[0036] 2.4 Uterine and Ovarian Coefficients and Histopathological Examination: Bilateral ovaries and uteruses were removed from each group of animals. Bloodstains were washed away with physiological saline, and the animals were blotted dry with filter paper before weighing. The organ coefficient was calculated (organ coefficient = organ weight ÷ body weight × 100%). The ovaries were fixed in formaldehyde, embedded, and prepared as slides. After HE staining, histological changes were observed under a light microscope.

[0037] After 28 days of drug administration, the uterine and ovarian coefficients of animals in each group were observed. Figure 4 The results showed no significant differences in uterine and ovarian coefficients among the drug-treated groups. Pathological results showed that the model control group had a large number of cystic dilated follicles in their ovaries, the low-dose Sanqi Danshen tablet group had a small number of cystic dilated follicles in their ovaries, and the normal control group, the high-dose Sanqi Danshen tablet group, and the metformin group had significantly fewer cystic dilated follicles in their ovaries (see...). Figure 5 ).

[0038] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. The application of Panax notoginseng and Salvia miltiorrhiza tablets in the preparation of medicines, characterized in that, The drug has at least one of the following effects: regulating hormone levels, alleviating insulin resistance, inhibiting cystic dilated follicles, promoting normal follicle development, and improving ovarian tissue structure.

2. The application according to claim 1, characterized in that, The regulation of hormone levels includes lowering the levels of at least one of the following hormones: testosterone, luteinizing hormone, anti-Müllerian hormone, and follicle-stimulating hormone.

3. Application of Panax notoginseng and Salvia miltiorrhiza tablets in the preparation of drugs for the treatment of polycystic ovary syndrome.

4. The application according to claim 3, characterized in that, The working dose of the Panax notoginseng and Salvia miltiorrhiza tablets in the drug is greater than 2.5 g / kg.

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