Application of sesquiterpene lactone compounds of Yemazhui in the preparation of drugs for treating polycystic ovary syndrome

Abstract

The present invention belongs to the field of biomedicine and specifically discloses the use of a sesquiterpene lactone compound from Yemazhui in the preparation of a drug for treating polycystic ovary syndrome. The use of the sesquiterpene lactone compound from Yemazhui in the preparation of a drug for treating polycystic ovary syndrome disclosed in the invention has the following beneficial effects: 1. For the first time, the sesquiterpene lactone compound from Yemazhui was applied to a mouse model of polycystic ovary syndrome. Related experiments demonstrated that the sesquiterpene lactone compound from Yemazhui alleviated polycystic ovary syndrome symptoms and inhibited ovarian and systemic inflammation; 2. The sesquiterpene lactone compound from Yemazhui has been clinically used, has a single component, and is safe and reliable, facilitating clinical application and promotion.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and particularly relates to the application of a sesquiterpene lactone compound of Yemazhui in the preparation of a drug for treating polycystic ovary syndrome. Background Art

[0002] Polycystic ovary syndrome (PCOS) is a relatively common endocrine disease in women of childbearing age, with a clinical incidence rate between 5% and 15%.

[0003] Clinically, PCOS often presents with elevated plasma free testosterone levels, oligomenorrhea, polycystic ovaries, infertility, hirsutism, insulin resistance, obesity, and acanthosis. The imbalance of estrogen and androgen in PCOS patients leads to the development of antral follicle cysts in the ovaries, which blocks the ovarian cycle and causes amenorrhea and infertility.

[0004] The pathogenesis of PCOS is still not fully understood. It is generally believed that PCOS is a gynecological disease caused by multiple genes and multiple factors. In addition to genetic and environmental factors, inflammatory factors are closely related to the pathogenesis of PCOS.

[0005] Regarding the treatment of PCOS, the current treatment is mainly aimed at metabolic-related symptoms such as high testosterone symptoms and insulin resistance. Treatment methods include taking progesterone and estrogen to inhibit excessive secretion of androgens, and taking metformin to alleviate insulin resistance symptoms. However, long-term use of hormonal drugs can cause serious side effects, and can only alleviate the symptoms of PCOS by controlling androgen levels, but cannot fundamentally cure PCOS. The current problems faced by patients with PCOS are long treatment cycles and the lack of specific drugs to treat PCOS.

[0006] Traditional Chinese medicine (TCM) is characterized by relatively good safety, relatively few side effects, and the ability to treat both the symptoms and the root cause. Herba Amaranthus chinensis (Herba Amaranthus) is the dried aerial part of the Asteraceae plant, Eupatorium verticillatum. It is bitter, neutral in nature, and has expectorant, cough-relieving, and asthma-relieving properties. Clinically, Herba Amaranthus chinensis is primarily used to treat chronic tracheitis and bronchitis. Herba Amaranthus chinensis contains natural compounds, and over 100 compounds have been identified from Herba Amaranthus chinensis, including terpenes, flavonoids, and alkaloids. Sesquiterpene lactones are the main characteristic and active ingredients. Studies have shown that Herba Amaranthus chinensis has anti-inflammatory, antioxidant, and anti-tumor pharmacological activities. Polycystic ovary syndrome (PCOS) is a type of inflammatory disease, and no studies have reported the use of Herba Amaranthus chinensis in the treatment of PCOS. Summary of the Invention

[0007] Purpose of the invention: In view of the shortcomings of the above-mentioned prior art, the present invention discloses the use of sesquiterpene lactone compounds of Yemazhui in the preparation of drugs for treating polycystic ovary syndrome.

[0008] Summary of the invention: Application of sesquiterpene lactone compounds of Yemazhui in the preparation of drugs for treating polycystic ovary syndrome.

[0009] Furthermore, the use of sesquiterpene lactone compounds of Yemazhui as the only active ingredient in the preparation of drugs for treating polycystic ovary syndrome.

[0010] Furthermore, the sesquiterpene lactone compound of YEMAZHUAI is YEMAZHUAI lactone A or YEMAZHUAI lactone B.

[0011] Furthermore, the drug is composed of a sesquiterpene lactone compound of Yemazhui and a pharmaceutically acceptable carrier.

[0012] Furthermore, the drug is in the form of any one of tablets, capsules, pills, powders, granules, and syrups.

[0013] Beneficial effects: The application of the Yemazhui sesquiterpene lactone compounds disclosed in the present invention in the preparation of drugs for treating polycystic ovary syndrome has the following beneficial effects:

[0014] 1. For the first time, the sesquiterpene lactone compounds from Yemazhui were applied to a mouse model of polycystic ovary syndrome. Experiments have shown that the sesquiterpene lactone compounds from Yemazhui can alleviate the symptoms of polycystic ovary syndrome and inhibit ovarian and systemic inflammation.

[0015] 2. The sesquiterpene lactone compounds of Yemazhui have been used clinically. They have a single ingredient and are safe and reliable, which is conducive to clinical application and promotion. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 Flowchart developed for the animal model.

[0017] Figure 2 Schematic diagram of the pathological section detection results of the ovarian sections of each group of mice after H&E staining, where: scale bar = 500 μm; Control represents the normal group; DHEA represents the control group; DHEA+EA represents the group injected with malachite lactone A; DHEA+EB represents the group injected with malachite lactone B.

[0018] Figure 3The figure is a schematic diagram of the statistical results of the number of corpora lutea in the ovarian sections of each group of mice after H&E staining, wherein: n=5, Control represents the normal group; DHEA represents the control group; DHEA+EA represents the group injected with malachite lactone A; DHEA+EB represents the group injected with malachite lactone B.

[0019] Figure 4 The figure is a schematic diagram of the statistical results of the number of cystic follicles in the ovarian sections of each group of mice after H&E staining, wherein: n=5, Control represents the normal group; DHEA represents the control group; DHEA+EA represents the group injected with yamachulide A; DHEA+EB represents the group injected with yamachulide B.

[0020] Figure 5 Schematic diagram of the glucose tolerance test (GTT) results of each group of mice, where: control group n=9, other groups n=5; Control represents the normal group; DHEA represents the control group; DHEA+EA represents the group injected with yamalide A; DHEA+EB represents the group injected with yamalide B.

[0021] Figure 6 Schematic diagram of the testosterone levels of mice in each group, where: n=5; Control represents the normal group; DHEA represents the control group; DHEA+EA represents the group injected with yamalide A; DHEA+EB represents the group injected with yamalide B.

[0022] Figure 7 CD4 + T / CD8 + Schematic diagram of the results of T cell ratio, where: Control represents the normal group; DHEA represents the control group; DHEA+EA represents the group injected with yamalide A; DHEA+EB represents the group injected with yamalide B. DETAILED DESCRIPTION

[0023] The specific embodiments of the present invention are described in detail below.

[0024] The "ranges" disclosed herein are defined in the form of lower and upper limits. A given range is defined by selecting a lower limit and an upper limit, and the selected lower and upper limits define the boundaries of the particular range. Ranges defined in this manner can be inclusive or exclusive and can be combined arbitrarily, i.e., any lower limit can be combined with any upper limit to form a range. For example, if a range of 10 to 50 is listed for a particular parameter, it is understood that ranges of 10 to 40 and 20 to 50 are also contemplated. Furthermore, if the minimum range values ​​listed are 1 and 2, and if the maximum range values ​​listed are 3, 4, and 5, then the following ranges are all contemplated: 1 to 3, 1 to 4, 1 to 5, 2 to 3, 2 to 4, and 2 to 5. In this application, unless otherwise specified, the numerical range "a to b" is an abbreviation for any combination of real numbers between a and b, where a and b are real numbers. For example, the numerical range "0 to 5" means that all real numbers between "0 to 5" are listed herein, and "0 to 5" is simply an abbreviation for these numerical combinations.

[0025] Unless otherwise specified, all embodiments and optional embodiments of the present application can be combined with each other to form a new technical solution.

[0026] Unless otherwise specified, all technical features and optional technical features of this application can be combined with each other to form a new technical solution.

[0027] Unless otherwise specified, all steps of the present application may be performed sequentially or randomly, preferably sequentially. For example, the method includes steps (a) and (b), indicating that the method may include steps (a) and (b) performed sequentially, or may include steps (b) and (a) performed sequentially. For example, the method may further include step (c), indicating that step (c) may be added to the method in any order, for example, the method may include steps (a), (b) and (c), or may include steps (a), (c) and (b), or may include steps (c), (a) and (b), etc.

[0028] Unless otherwise specified, the terms "include" and "comprising" used in this application may be open-ended or closed-ended. For example, "include" and "comprising" may mean that other components not listed may also be included or that only the listed components are included.

[0029] Unless otherwise specified, the reaction is carried out at room temperature and pressure.

[0030] Unless otherwise specified, all parts or percentages are by weight.

[0031] In the present invention, all substances used are known substances and can be purchased or synthesized by known methods.

[0032] In the present invention, the devices or equipment used are all conventional devices or equipment known in the art and are commercially available.

[0033] Experimental Materials:

[0034] 1. Mice: 3-week-old C57BL / 6J mice were purchased from SPIEF (Beijing) Biotechnology Co., Ltd. and raised under SPF-grade conditions. The mouse breeding conditions were consistent.

[0035] Experimental methods:

[0036] 1. Establishment of animal model.

[0037] Three-week-old female mice were randomly divided into a normal group, a control group, and a drug-treated group. The mice were kept in a constant temperature and humidity environment with free access to food and water. The experiment was carried out after one week of adaptation.

[0038] like Figure 1 As shown, mice in the control group and the treatment group were subcutaneously injected with DHEA at 0.18 mg / g body weight for 30 days. Mice in the normal group were subcutaneously injected with DHEA solution in sesame oil at 0.18 mg / g body weight for 30 days.

[0039] After 31 days, the mice in the treatment group were intragastrically administered with 20 mg / kg of yamalide A or 20 mg / kg of yamalide B. The mice in the normal group and the control group were intragastrically administered with 20 mg / kg of DHEA in sesame oil.

[0040] After 51 days, glucose tolerance, insulin tolerance, ovarian pathological sections and hormone levels of mice in each group were tested.

[0041] 2. Ovarian pathological biopsy.

[0042] Ovarian tissues were obtained from each group of mice to compare ovarian morphology. Ovarian tissues were fixed with 4% paraformaldehyde, embedded in paraffin, and sectioned. H&E staining was performed, and the number of corpora lutea and cystic follicles in each group was counted.

[0043] 3. Glucose tolerance test.

[0044] All mice in each group were fasted for 12 hours beforehand. Blood glucose levels were measured from the tail of the mice. Then, 2 g / kg of D-glucose was injected into the mice. Blood glucose levels were measured from the tail of the mice 15, 30, 60, 90, and 120 minutes after glucose injection.

[0045] 4. Insulin tolerance test.

[0046] All mice in each group were fasted for 4 hours beforehand. Blood glucose levels were measured by tail blood sampling, followed by an insulin injection at 1 IU / kg body weight. Blood glucose levels were measured by tail blood sampling 15, 30, 60, 90, and 120 minutes after injection.

[0047] 5. Androgen level testing.

[0048] 100 μl of mouse blood was collected from the medial canthus of the orbit and centrifuged at 3000 rpm for 10 min at 4°C. The supernatant serum was aspirated. Serum androgen levels were measured using an ELISA kit, following the manufacturer's instructions.

[0049] 6. Flow cytometric analysis.

[0050] 100 μl of blood was collected from the inner canthus of the orbit of each group of mice and centrifuged at 3000 rpm for 5 min to collect cells. After removing red blood cells with red blood cell lysis buffer, the cells were stained with CD4 and CD8 fluorescent antibodies, and the proportion of CD4-positive cells and CD8-positive cells was analyzed by flow cytometry.

[0051] Experimental results:

[0052] 1. A mouse model of polycystic ovary syndrome was established by subcutaneous injection of the androgen DHEA, and the mice in the treatment group were treated with 20 mg / kg of yesama lactone A or 20 mg / kg of yesama lactone B by oral gavage (e.g. Figure 1 shown).

[0053] 2. From Figure 2 It can be seen that compared with the mice in the untreated group (DHEA group), the ovarian bleeding symptoms of the mice in the 20 mg / kg Yesamalide A group (DHEA+EA) and the 20 mg / kg Yesamalide B group (DHEA+EB) were significantly improved.

[0054] 3. From Figure 3 The results of ovarian section staining showed that compared with the mice in the untreated group (DHEA group), the number of cystic follicles in the mice in the DHEA+EA group and the DHEA+EB group was significantly lower than that in the mice in the DHEA group.

[0055] 4. Figure 4 It can be seen that the number of corpora lutea in the mice in the DHEA+EA group and the DHEA+EB group was significantly higher than that in the mice in the DHEA group.

[0056] 5. Figure 5 The glucose tolerance test showed that insulin resistance was improved after treatment with yemachaylactone A or yemachaylactone B.

[0057] 6. Figure 6The testosterone level test results showed that the testosterone levels of the DHEA+EA and DHEA+EB groups were lower than that of the DHEA group.

[0058] 7. Flow cytometry detection of CD4 in peripheral blood of each group + T cells and CD8 + T cell ratio, Figure 7 The results showed that the CD4+ / CD8+T cell ratio of mice in the DHEA+EA and DHEA+EB groups was significantly lower than that in the DHEA group.

[0059] The above results show that after treatment with yesamalide lactone A or yesamalide lactone B, the symptoms of polycystic ovary syndrome in the polycystic ovary syndrome model group mice were improved and the inflammation level was reduced.

[0060] The embodiments of the present invention have been described in detail above, but the present invention is not limited to the above embodiments, and various modifications can be made within the scope of knowledge possessed by those skilled in the art without departing from the spirit of the present invention.

Claims

1. Use of a sesquiterpene lactone compound of Yemazhui in the preparation of a drug for treating polycystic ovary syndrome, wherein: The sesquiterpene lactone compound of YEMACHU is YEMACHU lactone A or YEMACHU lactone B.

2. The use according to claim 1, characterized in that The invention relates to the use of sesquiterpene lactone compounds of Yemazhui as the sole active ingredient in the preparation of a drug for treating polycystic ovary syndrome.

3. The use according to claim 1, characterized in that The medicine consists of a sesquiterpene lactone compound of Yemazhui and a pharmaceutically acceptable carrier.

4. The use according to claim 3, characterized in that The medicine is in the form of any one of tablets, capsules, pills, powders, granules and syrups.

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