补肾疗更浸膏治疗更年期综合征的机制研究

发布时间：2018-06-11 16:47

本文选题：补肾疗更浸膏 + 更年期综合征　；参考：《湖南中医药大学》2016年硕士论文

【摘要】：目的:通过研究中药新药-补肾疗更浸膏对去势更年期模型大鼠血清相关性激素、下丘脑单胺类神经递质、下丘脑β-内啡肽、自由基代谢与子宫组织病理学的影响,以及对老年小鼠血清相关性激素、卵巢储备功能、颗粒细胞凋亡与子宫、卵巢组织病理学的影响,探讨中药新药补肾疗更浸膏治疗更年期综合征的机制。方法:(1)通过手术摘除雌性SD大鼠双侧卵巢复制更年期模型,灌胃给药4周后腹主动脉取血,离心制备血清样本,采用ELISA方法检测模型大鼠血清雌二醇(E2)、促黄体生成素(LH)、促卵泡生成素(FSH)、促性腺激素释放激素(Gn RH)、睾酮(T)、丙二醛(MDA)的含量以及超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)的活性;分离下丘脑组织,制备组织匀浆,经BCA蛋白定量后采用ELISA方法检测模型大鼠下丘脑多巴胺(DA)、去甲肾上腺素(NE)、五羟色胺(5-HT)、五羟吲哚乙酸(5-HIAA)、β-内啡肽(β-EP)的含量;摘取双侧子宫,准确称量湿重计算脏器系数,石蜡包埋切片后HE染色,进行组织病理学观察并测量大鼠子宫内膜厚度。(2)采用老年小鼠复制更年期自然衰老模型,灌胃给药4周后眼内眦采血,分离制备血清样本,采用ELISA法检测老年小鼠血清E2、LH、FSH、孕酮(P)的含量;摘取双侧卵巢,准确称量湿重计算脏器系数,石蜡包埋切片,一部分卵巢经HE染色后进行组织病理学观察,并检测老年小鼠各级卵泡、黄体数,一部分卵巢通过DNA原位末端标记技术,采用TUNEL试剂盒检测老年小鼠卵巢颗粒细胞凋亡数;摘取双侧子宫,准确称量湿重计算脏器系数,石蜡包埋切片,HE染色观察其组织病理学改变并测量老年小鼠子宫内膜厚度。结果:(1)补肾疗更浸膏可有效升高去势模型大鼠血清E2含量(P0.05,P0.01),升高SOD、GSH-Px活性(P0.05,P0.01),降低血清LH、FSH、Gn RH、T、MDA含量(P0.05,P0.01);降低模型大鼠下丘脑5-HT、5-HIAA、DA、NE含量(P0.05,P0.01),升高β-EP含量(P0.05);增加模型大鼠子宫内膜厚度(P0.05),提高子宫系数(P0.05),缓解并改善子宫组织病理改变。(2)补肾疗更浸膏可有效升高老年小鼠血清E2、P含量(P0.05,P0.01),降低LH、FSH含量(P0.01);升高卵巢生长卵泡、成熟卵泡数(P0.05,P0.01);降低老年小鼠卵巢颗粒细胞凋亡数(P0.01);增加老年小鼠子宫内膜厚度(P0.05,P0.01),提高子宫、卵巢系数(P0.05,P0.01),缓解并改善子宫、卵巢组织病理改变。结论:中药新药-补肾疗更浸膏对更年期综合征具有明显的治疗作用,其机制主要为:(1)通过调节血液相关性激素含量,调节下丘脑-垂体-卵巢轴功能,改善机体内分泌,缓解更年期月经紊乱、月经不调、性欲冷淡等症状;(2)通过调节下丘脑单胺类神经递质含量,改善植物神经功能,缓解更年期失眠、烦躁、易怒等症状;(3)通过调节下丘脑内源性阿片肽含量,改善下丘脑神经内分泌功能以及体温调节中枢功能异常,缓解更年期月经周期紊乱,情志失调以及潮热等症状;(4)通过调节血液过氧化物酶活性以及自由基代谢产物含量,改善机体自由基代谢异常,缓解更年期组织器官氧化衰老,功能衰退;(5)通过调节卵巢各级卵泡、黄体数,增强卵巢储备功能,缓解更年期卵泡数量减少,质量减退;(6)通过抑制卵巢颗粒细胞凋亡,治疗更年期卵巢储备功能降低,卵泡多度耗竭;(7)通过改善子宫、卵巢组织微观形态,缓解子宫、卵巢组织病理性改变,恢复子宫、卵巢功能,改善生殖功能。
[Abstract]:Objective: To study the effects of new herbal medicine - tonifying kidney therapy on serum related hormones, monoamine neurotransmitters in hypothalamus, hypothalamus, beta endorphin, free radical metabolism and histopathology, as well as serum related irrigating, ovarian reserve, granulosa cell apoptosis and uterus and ovum in old mice. The mechanism of nesting histopathology was discussed. Methods: (1) the menopause model of bilateral ovary of female SD rats was removed by operation and the abdominal aorta was taken for 4 weeks. The serum samples were prepared by centrifugation, and the serum estradiol (E2) of the model rats was detected by ELISA. LH, follicle stimulating hormone (FSH), gonadotropin releasing hormone (Gn RH), testosterone (T), malondialdehyde (MDA), and the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px); the hypothalamus tissue was isolated and tissue homogenate was prepared. The rat hypothalamic DOPA was detected by ELISA method after BCA protein quantification. The hypothalamic DOPA was detected by ELISA method after BCA protein quantitative analysis. The content of amine (DA), norepinephrine (NE), five hydroxytryptamine (5-HT), five hydroxyindole acetic acid (5-HIAA), beta endorphin (beta -EP), extraction of bilateral uterus, accurate weighing wet weight calculation of organ coefficient, HE staining after paraffin embedded section, histopathological observation and measurement of endometrium thickness in rats. (2) the old mice were used to reproduce the natural decline of menopause. The old model, after 4 weeks of intragastric administration, the blood samples were collected in the eye canthus and the serum samples were prepared. The serum E2, LH, FSH, progesterone (P) content of the aged mice were detected by ELISA. The ovariectomized ovary was extracted and the viscera coefficient was accurately weighed, the paraffin embedded sections were embedded, some of the ovaries were observed histopathologically after HE staining, and the eggs of the aged mice were detected at all levels of eggs. Vesicles, corpus luteum and a part of ovary were detected by DNA in situ terminal labeling technique, TUNEL kit was used to detect the number of apoptosis in ovarian granulosa cells in old mice; bilateral uterus was extracted and the viscera coefficient was accurately weighed, paraffin embedded section, HE staining was used to observe histopathological changes and the thickness of endometrium in old mice. Results: (1) kidney tonifying. Increase the serum E2 content (P0.05, P0.01) in ovariectomized rats, increase SOD, GSH-Px activity (P0.05, P0.01), reduce the serum LH, FSH, Gn RH, T, increase the content of the rat's hypothalamus, increase the content of the beta endometrium, increase the thickness of the endometrium in the model rats; increase the thickness of the endometrium in the model rats, increase the thickness of the endometrium of the model rats, and increase the thickness of the endometrium of the model rats, and increase the thickness of the endometrium of the model rats, and increase the thickness of the endometrium of the model rats, and the increase of the endometrial thickness of the model rats, and the increase of the endometrial thickness of the model rats Uterine coefficient (P0.05), relieving and improving the histopathological changes of uterus. (2) TP can effectively increase serum E2, P content (P0.05, P0.01), decrease LH, FSH content (P0.01), increase ovarian follicle, mature follicle number (P0.05, P0.01), decrease the number of ovarian granulosa cell apoptosis (P0.01) in old mice and increase the uterus in old mice. Membrane thickness (P0.05, P0.01), improve the uterus, ovarian coefficient (P0.05, P0.01), relieve and improve the pathological changes of uterus and ovary. Conclusion: the new medicine - tonifying kidney therapy has obvious therapeutic effect on menopause syndrome. The mechanism is: (1) regulating the function of hypothalamus hypophysis ovary axis by regulating the blood related hormone content. Improve the body endocrine, menopause menstrual disorder, menstruation, lust and other symptoms; (2) through the regulation of the hypothalamus monoamine neurotransmitter content, improve plant nerve function, relieve menopause insomnia, irritability, irritability and other symptoms; (3) through the regulation of the hypothalamus endogenous opioid peptide content, improve the hypothalamus neuroendocrine function and Abnormal central function of thermoregulation, relieving menopause menstrual cycle disorders, emotional disorders and hot flashes, and (4) improving the metabolism of free radicals by regulating the activity of peroxidase and the content of free radical metabolites, relieving the aging of oxygen and function in climacteric organs and function decline; (5) by regulating ovarian follicles at all levels, yellow Body number, enhance ovarian reserve function, reduce the number of follicles in menopause, decrease the quality of follicle; (6) by inhibiting ovarian granulosa cell apoptosis, the treatment of ovarian granulosa cell apoptosis, treatment of ovarian reserve function decreased, follicle exhaustion; (7) through the improvement of the uterus, ovarian tissue micromorphology, the slow solution of uterus, ovarian tissue pathological changes, the restoration of uterus, ovarian function, improvement Reproductive function.
【学位授予单位】：湖南中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R271.116

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