Application of cannabidiol in preparation of medicine for treating polycystic ovarian syndrome
By using cannabidiol to intervene in the PCOS mouse model, the problem that existing drugs are difficult to improve multiple symptoms of PCOS at the same time is solved, and the effect of improving ovulation function, ovarian pathology, sugar metabolism, lipid metabolism and anxiety symptoms is achieved without side effects.
Patent Information
- Application Number
- CN202411847074.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-13
- Publication Date
- 2025-02-11
AI Technical Summary
Existing drugs for the treatment of polycystic ovarian syndrome (PCOS) are difficult to improve ovulation function, ovarian pathological status, sugar metabolism, lipid metabolism and anxiety symptoms at the same time, and are often accompanied by side effects, which limits long-term use.
Using cannabidiol (CBD) as the main component, the PCOS mouse model was intervened by subcutaneously embedding of silicone sustained-release tubes and continuous gavage, and regulated its ovulation function, ovarian pathological status, sugar metabolism, lipid metabolism and anxiety-like behavior. .
The ovulation function of PCOS mice was significantly restored, the ovarian pathological status was improved, the sugar and lipid metabolic disorders were reduced, anxiety-like behavior was improved, and there was no psychoactive and drug dependence.
Smart Images
Figure CN119405635A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the field of medicine, and specifically relates to the application of cannabidiol in preparing a medicine for treating polycystic ovary syndrome. Background Art
[0002] Polycystic ovary syndrome (PCOS) is a common complex endocrine syndrome in women of childbearing age. It is the leading cause of ovulatory infertility, with an incidence rate of 6-19% in women of childbearing age. PCOS is characterized by chronic anovulation (ovulatory dysfunction or loss), hyperandrogenism, and polycystic ovaries, but PCOS also covers many clinical features, including acne, hirsutism, alopecia, menstrual disorders, infertility, and mood disorders in the short term, and metabolic disorders in the long term, increasing the risk of hepatic steatosis, type 2 diabetes, metabolic syndrome, and cardiovascular disease.
[0003] At present, the clinical treatment of PCOS mainly focuses on symptomatic treatment such as anti-androgen, ovulation induction, and insulin sensitization. Few drugs can effectively target all aspects of PCOS. Combined oral contraceptives (COCs) are recommended for the treatment of hyperandrogenism and / or irregular menstrual cycles in adult women with PCOS. However, COCs do not improve infertility, polycystic ovary morphology, and mood disorders. In addition, COCs are often accompanied by side effects such as thromboembolism and mood swings, which limit the long-term clinical application of COCs, especially for PCOS patients with metabolic disorders.
[0004] Cannabidiol (CBD) is a cannabinoid extracted from cannabis. It has very low affinity with cannabinoid type 1 receptor (CB1R) and type 2 receptor (CB2R), and is not psychoactive or drug-dependent, so it is relatively safe. As the main cannabinoid extract, CBD is believed to have anticonvulsant, antianxiety, antipsychotic and neuroprotective effects. Subsequently, it has been used for metabolic diseases such as obesity, fatty liver and insulin resistance because of its anti-inflammatory and anti-oxidative stress effects. However, cannabidiol has not yet been used to treat PCOS. Summary of the invention
[0005] Purpose of the invention: The purpose of the present invention is to overcome the shortcomings of the prior art and, based on the existing efficacy of cannabidiol, screen its other clinical uses through a large number of experiments.
[0006] The present invention has found through experiments that cannabidiol can significantly restore ovulation function (including improving serum progesterone levels and regulating the estrous cycle of mice), improve the ovarian pathological state of PCOS mice (such as reducing the number of cystic atretic follicles in ovarian tissue and increasing the number of corpora lutea), and can also improve the sugar and lipid metabolism of PCOS mice and improve the anxiety-like behavior of PCOS mice.
[0007] Therefore, the present invention proposes the use of cannabidiol in the preparation of a medicine for treating polycystic ovary syndrome.
[0008] Application of cannabidiol in the preparation of medicines for improving polycystic ovary syndrome complicated with glucose metabolism disorder.
[0009] Application of cannabidiol in the preparation of drugs for improving lipid metabolism disorders associated with polycystic ovary syndrome.
[0010] Application of cannabidiol in the preparation of medicines for polycystic ovary syndrome complicated with anxiety disorder.
[0011] The beneficial effects achieved by the present invention are as follows:
[0012] (1) The present invention discloses the use of CBD in the preparation of a drug for treating PCOS. CBD produces a pharmacological effect by regulating the estrous cycle of mice, improving the ovarian pathological state of PCOS mice, improving the sugar and lipid metabolism of PCOS mice, and improving the anxiety-like behavior of PCOS mice.
[0013] (2) The present invention constructs a high androgenic PCOS mouse model by subcutaneous embedding of DHT. Hematoxylin / eosin staining is used to observe the mouse vaginal exfoliated cells to determine the estrous cycle of the mouse. The mouse shows an interrupted estrous cycle. After administration of CBD, the estrous cycle of the PCOS mouse is restored.
[0014] (3) Reduce the number of cystic atretic follicles and increase the number of corpora lutea in ovarian tissue to improve PCOS.
[0015] (4) Reduce glucose tolerance impairment and improve glucose metabolism.
[0016] (5) Reduce serum cholesterol levels, improve the accumulation of lipid droplets in white adipocytes and brown adipocytes near the gonads of PCOS mice, reduce cell area, and improve lipid metabolism
[0017] (6) In the open field test, CBD significantly increased the time and frequency of PCOS mice entering the central area. In the elevated plus maze test, CBD significantly increased the time and frequency of PCOS mice entering the open arms, and improved the anxiety-like behavior of PCOS mice. BRIEF DESCRIPTION OF THE DRAWINGS
[0018] Figure 1CBD improves reproductive function in DHT-induced PCOS mice;
[0019] Figure 2 CBD improves lipid metabolism in DHT-induced PCOS mice;
[0020] Figure 3 CBD improves glucose metabolism in DHT-induced PCOS mice;
[0021] Figure 4 CBD improves DHT-induced anxiety-like behavior in PCOS mice. DETAILED DESCRIPTION
[0022] Example 1
[0023] 1. Materials and methods
[0024] 1.1 Construction of androgen-induced PCOS mouse model
[0025] Control group: Empty silicone sustained-release tubes were subcutaneously implanted in the back of the neck of 3-week-old SPF female C57BL / 6 mice for 90 days. PCOS model group: Silicone sustained-release tubes containing 10 mg dihydrotestosterone (DHT) were subcutaneously implanted in the back of the neck of 3-week-old SPF female C57BL / 6 mice for 90 days. Cannabidiol (CBD) intervention group (low-dose group, high-dose group): Silicone sustained-release tubes containing 10 mg DHT were subcutaneously implanted in the back of the neck of 3-week-old SPF female C57BL / 6 mice for 90 days. After 62 days of implantation, CBD was dissolved in 1% sodium carboxymethylcellulose and gavaged for 4 weeks. After 62 days of implantation, the control group and model group silicone sustained-release tubes were gavaged with an equal volume of 1% sodium carboxymethylcellulose for 4 weeks. The CBD doses (100 mg / kg / d and 200 mg / kg / d) for PCOS mice in this study were determined according to the literature (Xu C, Chang T, Du Y, Yu C, Tan X, Li X. Pharmacokinetics of oral and intravenous cannabidiol and its antidepressant-like effects in chronic mild stress mouse model. Environ Toxicol Pharmacol. 2019; 70: 103202.). The animal experimental design and procedures were in accordance with the requirements of the Animal Ethics Committee of Nanjing University of Chinese Medicine and were conducted in accordance with the Declaration of Helsinki. 1.2 Effects of CBD intervention on the estrous cycle of PCOS model mice
[0026] The estrous cycle of mice was monitored by vaginal smear and HE staining, and the smears were continued for 2 weeks. In the proestrus, all cells were nucleated epithelial cells, with occasional small amounts of keratinized cells. In the estrus, all cells were keratinized cells, with occasional small amounts of epithelial cells. In the metestrus, all cells were keratinized epithelium, leukocytes, and nucleated epithelial cells. In the diestrus, there were a large number of leukocytes and a small number of epithelial cells and mucosa.
[0027] 1.3 Effect of CBD intervention on ovarian tissue morphology in PCOS model mice
[0028] Ovaries were obtained from mice in diestrus, fixed with 4% paraformaldehyde, and sliced in paraffin. The pathological morphology of the ovaries was observed by HE staining. The number of atretic follicles and corpora lutea were counted.
[0029] 1.4 Effects of CBD intervention on serum sex hormones in PCOS model mice
[0030] After the blood was collected from the mice, it was left to stand for 30 minutes, centrifuged at 3000 g for 10 minutes, and the supernatant was collected. The serum estradiol (E2), progesterone (P), testosterone (T), and DHT levels were detected by ELISA according to the kit instructions.
[0031] 1.4 Effect of CBD intervention on glucose metabolism in PCOS model mice
[0032] Glucose tolerance test: At the end of the experiment, the mice were fasted for 12 hours and then injected with glucose solution at 1.5 g / kg body weight intraperitoneally. The tail vein blood glucose level of each group was measured before injection and 30 minutes, 60 minutes, 90 minutes and 120 minutes after injection using a rapid whole blood glucose tester.
[0033] 1.5 Effect of CBD intervention on lipid metabolism in PCOS model mice
[0034] The levels of serum triglyceride and cholesterol in each group of mice were detected by biochemical method.
[0035] 1.6 Effects of CBD intervention on the behavior of PCOS model mice
[0036] The open field test and elevated plus maze test are commonly used methods to evaluate the anxiety-like behavior of experimental animals. 90 days after DHT exposure, the open field test and elevated plus maze test were used to test the behavior of mice.
[0037] Open field test: The whole experiment was conducted in dark conditions. The mice were acclimated in the dark behavioral analysis room for 3-4 hours before the experiment. Before each mouse was tested, the instrument was wiped with 75% alcohol solution to eliminate olfactory cues. The mouse was placed in the center of the open field with its back facing the experimenter. The video analysis system recorded the movement time of the central area and the number of times it entered the central area, and the total recording time was 5 minutes.
[0038] Elevated plus maze test: The experimental environment and animal adaptation time of the elevated plus maze are the same as those of the open field test. At the beginning of the test, the mouse is placed on the central platform of the elevated plus maze with its back to the experimenter, facing one of the open arms. The video analysis system records the time the mouse enters the open arm movement time, the closed arm movement time, the number of arm openings, and the number of arm closings, and calculates the open / total time percentage (OT%) = open arm movement time / (open arm movement time + closed arm movement time) × 100%, open / total number percentage (OE%) = open arm number / (open arm number + closed arm number) × 100%. The time for each group of mice to perform the elevated plus maze test is the same as the open field test.
[0039] Now will refer to the accompanying drawings to describe in detail various exemplary embodiments of the present invention, the features and advantages of the present invention will become clear. The following description of at least one exemplary embodiment is only illustrative in nature and is not intended to limit the present invention and its application or use.
[0040] 2. Experimental results
[0041] 2.1 CBD improves reproductive function in DHT-induced PCOS mice
[0042] like Figure 1 The experimental results showed that the serum DHT level of PCOS mice induced by DHT increased significantly, and the estrous cycle was interrupted. The granulosa cells in the follicles of PCOS mice were loose, the number of layers decreased, the follicles were poorly developed, the number of atretic follicles increased, and the number of corpora lutea decreased after ovulation. The serum progesterone level of PCOS mice was significantly reduced. Compared with DHT-induced PCOS mice, CBD can improve the interruption of the estrous cycle of PCOS mice, increase the number of granulosa cell layers in the follicles, improve follicle development, reduce the number of atretic follicles, and increase the number of corpora lutea after ovulation. The serum progesterone level of PCOS mice increased significantly.
[0043] 2.2 CBD improves lipid metabolism in DHT-induced PCOS mice
[0044] like Figure 2 The experimental results showed that although the serum triglyceride level of DHT-induced PCOS mice was not significantly different from that of the normal control group, the serum cholesterol level of DHT-induced PCOS mice was significantly increased. Compared with DHT-induced PCOS mice, high and low doses of CBD significantly reduced the serum cholesterol level of PCOS mice. Moreover, the accumulation of lipid droplets in white adipocytes and brown adipocytes around the gonads of DHT-induced PCOS mice increased, and the cell area increased. CBD improved the accumulation of lipid droplets in white adipocytes and brown adipocytes around the gonads of PCOS mice, and the cell area decreased.
[0045] 2.3 CBD improves glucose metabolism in DHT-induced PCOS mice
[0046] like Figure 3 The experimental results showed that although there was no significant difference in the fasting blood glucose level between DHT-induced PCOS mice and the normal control group, in the glucose tolerance test, the glucose area under the curve (AUC) of DHT-induced PCOS mice was significantly increased. Compared with DHT-induced PCOS mice, high-dose CBD significantly reduced the glucose AUC of PCOS mice.
[0047] 2.4 CBD improves DHT-induced anxiety-like behavior in PCOS mice
[0048] like Figure 4 The experimental results showed that in the open field test, the time and number of times that DHT-induced PCOS mice entered the central area were reduced. Compared with DHT-induced PCOS mice, high-dose CBD significantly increased the time and number of times that PCOS mice entered the central area. In the elevated plus maze test, the time and number of times that DHT-induced PCOS mice entered the open arms were reduced. Compared with DHT-induced PCOS mice, high-dose CBD significantly increased the time and number of times that PCOS mice entered the open arms, which has an anti-anxiety effect.
[0049] The embodiments of the present invention are provided for the purpose of illustration and description, and are not intended to be exhaustive or to limit the present invention to the disclosed forms. Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for those skilled in the art to modify the technical solutions described in the aforementioned embodiments, or to make equivalent substitutions for some of the technical features therein.
Claims
1. The application of cannabidiol in the preparation of medicines for the treatment of polycystic ovary syndrome.
2. The use according to claim 1, characterized in that The cannabidiol is used in the preparation of drugs for restoring mammalian ovulation function and improving ovarian tissue pathological changes.
3. The use according to claim 2, characterized in that The indicators of ovulatory function include estrous cycle and serum progesterone level; The indicators of ovarian histological changes include the number of atretic follicles and corpora lutea in ovarian tissue.
4. The use according to claim 1, characterized in that The cannabidiol is used in the preparation of a medicine for improving polycystic ovary syndrome complicated with glucose metabolism disorder.
5. The use according to claim 1, characterized in that The cannabidiol is used in the preparation of a drug for improving lipid metabolism disorder complicated by polycystic ovary syndrome.
6. The use according to claim 1, characterized in that The application of cannabidiol in preparing medicine for polycystic ovary syndrome complicated with anxiety disorder.
7. A drug for treating polycystic ovary syndrome, characterized in that: Including cannabidiol.