身心应激诱导大鼠卵巢早衰模型的建立及机制研究

发布时间：2018-03-11 01:08

  本文选题：身心应激　切入点：卵巢早衰　出处：《华中科技大学》2010年硕士论文　论文类型：学位论文

【摘要】：目的:通过给予雌性SD大鼠长期的身心应激,建立大鼠POF模型,观察大鼠动情周期、行为症状改变、神经内分泌的改变以及卵巢局部因子的变化,研究身心应激引起POF的机制。 方法:将SD雌性大鼠随机分为正常组和模型组。正常组不做任何处理,常规喂养。模型组采用改良国际应激方法,每天随机抽取一种方法给予应激,应激每一笼模型组大鼠时,将其余模型组大鼠放于旁边,共同处于应激气氛中,以达到心理应激的目的。模型组连续应激28天,其间观察大鼠行为及最后十天观察动情周期变化,于第29天时同时处死正常组和模型组,收集大鼠卵巢、血清以及脑组织,采用HE染色法观测卵巢病理学改变,免疫组织化学法测定卵巢bax和VEGF蛋白的含量,放免法测定血清FSH和E2的含量,ELISA法检测NE、皮质酮和INHB的含量以及Realtime-PCR法检测卵巢GDF-9B mRNA的表达。 结果:行为症状显示,与正常组相比,模型组大鼠皮毛枯燥易掉,行动减少、呆滞,抓咬减少,洗脸次数增多,体重减轻。与正常组相比,HE染色卵巢病理学改变显示模型组大鼠各级正常卵泡明显减少(P0.05),闭锁卵泡显著增多(P0.01),黄体数目显著减少(P0.01),且体积大小不均;与正常组相比,模型组神经内分泌改变显示血清FSH含量升高(P0.01),E2含量降低(P0.05);模型组下丘脑NE值有下降趋势(P0.05),垂体NE值降低(P0.05),血清NE值升高(P0.01);血清皮质酮显著升高(P0.01);血清INHB显著降低(P0.01);与正常组相比,模型组卵巢bax蛋白表达高于正常组(P0.01);VEGF蛋白低于正常组(P0.01);模型组卵巢GDF-9B mRNA表达有下降趋势,但差异无统计学意义(P0.05)。 结论: 1 .身心应激能导致部分雌鼠卵巢早衰,是卵巢早衰的重要病因之一。 2.身心应激可引起精神-神经-内分泌紊乱以致神经递质分泌失调,导致下丘脑-垂体-肾上腺轴和下丘脑-垂体-卵巢轴破坏失衡,导致卵巢早衰。 3.身心应激可下调雌鼠卵巢血管内皮生长因子,减少卵巢血管发生,使卵巢血供减少,改变卵巢细胞内环境,增加颗粒细胞凋亡因子bax表达,影响卵母细胞GDF-9B mRNA表达,使卵泡闭锁加速,最终共同作用导致卵巢早衰。
[Abstract]:Objective: to study the mechanism of POF induced by physical and mental stress in female SD rats by observing the estrous cycle, behavioral changes, neuroendocrine changes and changes of local factors of ovary. Methods: SD female rats were randomly divided into normal group and model group. When each cage model group rats were stressed, the rest of the model group rats were placed on the side, together in the stress atmosphere, in order to achieve the purpose of psychological stress. The model group was under continuous stress for 28 days, and the rats in the model group were subjected to stress for 28 days. The behavior of rats and the changes of estrus cycle in the last ten days were observed. On the 29th day, the normal group and the model group were killed at the same time, the ovary, serum and brain tissues were collected, and the pathological changes of the ovary were observed by HE staining. The contents of bax and VEGF protein in ovary were detected by immunohistochemistry, the contents of FSH and E2 in serum were detected by radioimmunoassay, the contents of INHB and corticosterone were detected by Elisa, and the expression of GDF-9B mRNA in ovary was detected by Realtime-PCR method. Results: the behavioral symptoms showed that compared with the normal group, the fur of the model group was dull and easy to lose, the movement decreased, the grip decreased, the number of facial washing increased. Compared with the normal group, the pathological changes of the ovary stained by HE staining showed that the normal follicles in the model group decreased significantly (P 0.05), the atresia follicles increased significantly (P 0.01), the number of corpus luteum decreased significantly (P 0.01), and the volume was uneven, compared with the normal group. The neuroendocrine changes in the model group showed that the serum FSH content was increased P0.01E _ 2 content was decreased, the hypothalamus NE value was decreased, the pituitary NE value was decreased, the serum NE value was increased, the serum NE value was increased, the serum corticosterone level was significantly increased, and the serum INHB was significantly decreased. Compared with normal group, The expression of bax protein in the model group was higher than that in the normal group, and the expression of GDF-9B mRNA in the model group was lower than that in the normal group, but there was no significant difference in the expression of GDF-9B mRNA between the model group and the normal group (P 0.05). Physical and mental stress can lead to premature ovarian failure in some female rats, which is one of the important causes of premature ovarian failure. 2. 2. Mental and physical stress can lead to the disorder of neurotransmitter secretion, the destruction and imbalance of hypothalamus-pituitary-adrenal axis and hypothalamus-pituitary-ovarian axis, and premature ovarian failure. 3. Physical and mental stress could down-regulate vascular endothelial growth factor (VEGF), decrease ovarian angiogenesis, decrease ovarian blood supply, change the intracellular environment of ovary, increase the expression of apoptotic factor bax in granulosa cells, and affect the expression of GDF-9B mRNA in oocytes. Accelerate follicular atresia and eventually lead to premature ovarian failure.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R-332

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