Application of lindley eupatorium sesquiterpene lactone compound in preparation of medicine for treating polycystic ovarian syndrome

Abstract

The invention belongs to the field of biological medicines, and particularly discloses application of a lindley eupatorium sesquiterpene lactone compound in preparation of a medicine for treating polycystic ovarian syndrome. The application of the lindley eupatorium sesquiterpene lactone compound in preparing the medicine for treating the polycystic ovarian syndrome disclosed by the invention has the following beneficial effects: 1, the lindley eupatorium sesquiterpene lactone compound is applied to a mouse model with the polycystic ovarian syndrome for the first time; related experiments prove that the lindley eupatorium sesquiterpene lactone compound has the effects of relieving the symptoms of the polycystic ovarian syndrome and inhibiting ovarian and systemic inflammation; 2, the lindley eupatorium sesquiterpene lactone compound is applied clinically, is single in component, is safe and reliable, and is beneficial to clinical application and popularization.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and particularly relates to the application of eupatorium lindleyanum sesquiterpene lactones in the preparation of drugs for treating polycystic ovary syndrome. Background Art

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

[0003] Clinically, polycystic ovary syndrome often manifests as excessive plasma free testosterone levels, oligomenorrhea, polycystic ovaries, infertility, hirsutism, insulin resistance, obesity, and acanthosis. The imbalance of estrogen and androgen in the body of patients with polycystic ovary syndrome leads to the formation of antral follicle cysts in the ovaries, blocking the periodic ovulation activity of the ovaries and resulting in amenorrhea and infertility in women.

[0004] Regarding the pathogenesis of polycystic ovary syndrome, it is not yet fully understood. Generally, polycystic ovary syndrome is a gynecological disease induced by multiple genes and multiple factors. In addition to being related to genetic factors and environmental factors, inflammatory factors are closely related to the pathogenesis of polycystic ovary syndrome.

[0005] Regarding the treatment of polycystic ovary syndrome, currently, it mainly focuses on treating hyperandrogenism symptoms and metabolic-related symptoms such as insulin resistance. The treatment methods include taking progesterone and estrogen to inhibit the excessive secretion of androgens, and taking metformin to relieve insulin resistance symptoms. However, long-term use of hormonal drugs for treatment will bring serious side effects, and it can only relieve the symptoms of polycystic ovary syndrome by controlling androgen levels, and cannot fundamentally treat polycystic ovary syndrome. The current problems faced by the treatment of patients with polycystic ovary syndrome are the long treatment cycle and the lack of specific drugs for treating polycystic ovary syndrome.

[0006] Traditional Chinese medicine has the characteristics of relatively good safety, relatively small side effects, and treating both the symptoms and the root causes. Eupatorium lindleyanum is the dried aerial part of the plant Eupatorium lindleyanum DC. of the Compositae family. It is bitter in taste and neutral in nature, and has the effects of resolving phlegm, relieving cough, and calming asthma. Clinically, Eupatorium lindleyanum is mainly used to treat chronic tracheitis and bronchitis. Eupatorium lindleyanum contains natural compounds. Currently, more than 100 compounds have been identified from Eupatorium lindleyanum, including terpenoids, flavonoids, and alkaloids, etc. Among them, sesquiterpene lactones are the main characteristic components and active components. Research shows that Eupatorium lindleyanum has pharmacological activities such as anti-inflammatory, antioxidant, and anti-tumor. Polycystic ovary syndrome is a kind of inflammation-related disease, and there has been no research report on the application of Eupatorium lindleyanum in the treatment of polycystic ovary syndrome. Summary of the Invention

[0007] Objective of the Invention: Aiming at the deficiencies existing in the above-mentioned prior art, the present invention discloses the application of eupatorium lindleyanum sesquiterpene lactone compounds in the preparation of drugs for treating polycystic ovary syndrome.

[0008] Content of the Invention: The application of eupatorium lindleyanum sesquiterpene lactone compounds in the preparation of drugs for treating polycystic ovary syndrome.

[0009] Furthermore, the application of eupatorium lindleyanum sesquiterpene lactone compounds as the sole active ingredient in the preparation of drugs for treating polycystic ovary syndrome.

[0010] Furthermore, the eupatorium lindleyanum sesquiterpene lactone compounds are eupalactone A or eupalactone B.

[0011] Furthermore, the drug is composed of eupatorium lindleyanum sesquiterpene lactone compounds and a pharmaceutically acceptable carrier.

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

[0013] Beneficial Effects: The application of the eupatorium lindleyanum sesquiterpene lactone compounds disclosed by 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 eupatorium lindleyanum sesquiterpene lactone compounds are applied to a polycystic ovary syndrome mouse model, and relevant experiments prove that the eupatorium lindleyanum sesquiterpene lactone compounds have the effects of relieving the symptoms of polycystic ovary syndrome and inhibiting ovarian and systemic inflammation;

[0015] 2. The eupatorium lindleyanum sesquiterpene lactone compounds have been clinically applied, with a single component and reliable safety, which is conducive to clinical application and promotion. Description of the Drawings

[0016] Figure 1 It is a flow chart for establishing an animal model.

[0017] Figure 2 It is a schematic diagram of the pathological section detection results of ovarian sections of each group of mice after H&E staining, where: scale = 500μm; Control represents the normal group; DHEA represents the control group; DHEA + EA represents the drug administration group injected with eupalactone A; DHEA + EB represents the drug administration group injected with eupalactone B.

[0018] Figure 3Schematic diagram of the statistical results of the number of corpora lutea in the ovarian sections of mice in each group after H&E staining, where: n = 5, Control represents the normal group; DHEA represents the control group; DHEA+EA represents the administration group injected with eupatoriopicrin A; DHEA+EB represents the administration group injected with eupatoriopicrin B.

[0019] Figure 4 Schematic diagram of the statistical results of the number of cystic follicles in the ovarian sections of mice in each group after H&E staining, where: n = 5, Control represents the normal group; DHEA represents the control group; DHEA+EA represents the administration group injected with eupatoriopicrin A; DHEA+EB represents the administration group injected with eupatoriopicrin B.

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

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

[0022] Figure 7 For CD4 + T / CD8 + Schematic diagram of the results of the T cell ratio in the peripheral blood of mice in each group, where: Control represents the normal group; DHEA represents the control group; DHEA+EA represents the administration group injected with eupatoriopicrin A; DHEA+EB represents the administration group injected with eupatoriopicrin B. Detailed implementation manners

[0023] The following details the specific implementation manners of the present invention.

[0024] The "ranges" disclosed in the present invention are defined in the form of a lower limit and an upper limit. A given range is defined by selecting a lower limit and an upper limit, and the selected lower limit and upper limit define the boundaries of a particular range. The ranges defined in this way can include the end values or not include the end values, and can be combined arbitrarily, that is, any lower limit can be combined with any upper limit to form a range. For example, if a range of 10-50 is listed for a specific parameter, it is understood that ranges of 10-40 and 20-50 are also contemplated. In addition, if the minimum range values 1 and 2 are listed, and if the maximum range values 3, 4, and 5 are listed, then the following ranges are all contemplated: 1-3, 1-4, 1-5, 2-3, 2-4, and 2-5. In this application, unless otherwise specified, the numerical range "a-b" represents an abbreviated representation of any real number combination between a and b, where a and b are both real numbers. For example, the numerical range "0-5" means that all real numbers between "0-5" have been fully listed herein, and "0-5" is only an abbreviated representation of these numerical combinations.

[0025] Unless otherwise specified, all embodiments and alternative embodiments of this application can be combined with each other to form new technical solutions.

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

[0027] Unless otherwise specified, all steps of this application can be carried out in sequence or randomly, preferably in sequence. For example, the method includes steps (a) and (b), which means that the method can include steps (a) and (b) carried out in sequence, or can also include steps (b) and (a) carried out in sequence. For example, it is mentioned that the method may further include step (c), which means that step (c) can be added to the method in any order. For example, the method can include steps (a), (b), and (c), or can also include steps (a), (c), and (b), or can also include steps (c), (a), and (b), etc.

[0028] Unless otherwise specified, the "including" and "comprising" mentioned in this application mean open-ended or can also be closed-ended. For example, the "including" and "comprising" can mean that other components not listed can also be included or comprised, or can only include or comprise the listed components.

[0029] Unless otherwise specified, the reaction is carried out under normal temperature and normal pressure conditions.

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

[0031] In the present invention, the substances used are all known substances, which 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 can be purchased.

[0033] Experimental materials:

[0034] 1. Mice: 3-week-old C57BL / 6J mice, purchased from Beijing SPF Biotechnology Co., Ltd., and raised under SPF-level breeding conditions with consistent breeding conditions for mice.

[0035] Experimental methods:

[0036] 1. Establishment of animal models.

[0037] Three-week-old female mice were randomly divided into a normal group, a control group, and a drug administration group. The breeding environment was kept at a constant temperature and humidity, and the mice were allowed to eat and drink freely. After one week of adaptation, the experiment was carried out.

[0038] As Figure 1 shown, the control group mice and the drug administration group mice were subcutaneously injected with DHEA at a dose of 0.18 mg / g body weight for 30 consecutive days. The normal group mice were subcutaneously injected with a DHEA sesame oil solution at a dose of 0.18 mg / g body weight for 30 consecutive days.

[0039] On the 31st day, the drug administration group mice were given 20 mg / kg Eupatorium lactone A or 20 mg / kg Eupatorium lactone B by gavage. The normal group mice / control group mice were given 20 mg / kg DHEA sesame oil solution by gavage.

[0040] On the 51st day, glucose tolerance, insulin tolerance, ovarian pathological section, and hormone level detection were performed on each group of mice.

[0041] 2. Detection of ovarian pathological sections.

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

[0043] 3. Glucose tolerance test.

[0044] Each group of mice was fasted for 12 hours in advance. The blood glucose content of the mice was detected by collecting tail blood, and then D-glucose was injected at a dose of 2 g / kg body weight. At 15 min, 30 min, 60 min, 90 min, and 120 min after glucose injection, the tail blood of each group of mice was collected to detect the blood glucose content.

[0045] 4. Insulin tolerance test.

[0046] Fasted the mice in each group 4 hours in advance. Collected the tail blood of the mice to detect the blood glucose content, and then injected insulin at 1 IU / kg body weight. At 15 min, 30 min, 60 min, 90 min, and 120 min after injection, collected the tail blood of the mice to detect the blood glucose content respectively.

[0047] 5. Detection of androgen level.

[0048] Collected 100 μl of blood from the inner canthus of the mouse orbit, centrifuged at 3000 rpm for 10 min at 4 °C, and aspirated the upper serum. Detected the androgen level in the serum using an ELISA kit, and strictly operated according to the instructions of the ELISA kit.

[0049] 6. Flow cytometry analysis.

[0050] Collected 100 μl of blood from the inner canthus of the mouse orbit in each group, centrifuged at 3000 rpm for 5 min, collected the cells, removed the red blood cells with red blood cell lysate, stained with CD4 and CD8 fluorescent antibodies, and analyzed the proportions of CD4-positive cells and CD8-positive cells by flow cytometry.

[0051] Experimental results:

[0052] 1. Established a polycystic ovary syndrome mouse model by subcutaneous injection of androgen DHEA, and treated the mice in the administration group with 20 mg / kg eupatoriopicrin A or 20 mg / kg eupatoriopicrin B by gavage (as 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 eupatoriopicrin A group (DHEA+EA) and the 20 mg / kg eupatoriopicrin B group (DHEA+EB) were significantly improved.

[0054] 3. From Figure 3 the staining results of ovarian sections 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 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 DHEA group.

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

[0057] 6. Figure 6The testosterone level detection results show that the testosterone levels in the DHEA+EA and DHEA+EB groups are lower than those in the DHEA group.

[0058] 7. Flow cytometry was used to detect the proportions of CD4 + T cells and CD8 + T cells in the peripheral blood of each group. Figure 7 The results show that the proportion of CD4+ / CD8+ T cells in the mice of the DHEA+EA and DHEA+EB groups is significantly lower than that in the DHEA group.

[0059] Taken together, the above results indicate that after treatment with eupatoriopicrin A or eupatoriopicrin B, the symptoms of polycystic ovary syndrome in the mice of the polycystic ovary syndrome model group are improved and the inflammation level is reduced.

[0060] The above has made a detailed description of the embodiments of the present invention. However, the present invention is not limited to the above embodiments, and various changes can be made without departing from the gist of the present invention within the knowledge scope of those of ordinary skill in the art.

Claims

1. Application of sesquiterpene lactone compounds of YEMAZHUAI in the preparation of drugs for the treatment of polycystic ovary syndrome.

2. The use according to claim 1, characterized in that Application of sesquiterpene lactone compounds of Yemazhui as the only active ingredient in the preparation of drugs for treating polycystic ovary syndrome.

3. The use according to claim 1 or 2, characterized in that: The sesquiterpene lactone compound of Herba Lycopersici is Herba Lycopersici lactone A or Herba Lycopersici lactone B.

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

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

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