慢性应激对卵泡发育的影响及自分泌、旁分泌调控机制研究

发布时间：2018-08-24 12:37

【摘要】： 目的: 观察慢性应激对卵泡发育和动情周期的影响及其对卵泡发育的非促性腺激素依赖的影响。选取GDF-9、BMP-15、bFGF、SCF四个代表性的卵巢旁分泌、自分泌因子行RT-PCR,观察慢性应激是否影响这些调控卵泡发育的卵巢局部调控因子的表达。方法: (1)通过建立小鼠慢性应激模型,制备卵巢HE切片观察慢性应激对卵泡发育的影响。 (2)从慢性应激的第一天开始(至处死日止)取小鼠阴道分泌物,作涂片,行瑞氏染色观察其所处动情周期。 (3)通过外源性补充促性腺激素,观察其对应激后卵泡发育的恢复作用。 (4)采用相对定量real-time PCR方法测定GDF-9、BMP-15、bFGF、SCF这几个代表性的卵巢旁分泌、自分泌因子的mRNA水平。实验结果: (1)慢性应激使小鼠的动情周期紊乱、间期延长。 (2)慢性应激影响卵泡的发育及排卵的发生。 (3)外源性补充促性腺激素不能使卵泡发育和排卵完全恢复正常。 (4)卵源性自分泌、旁分泌因子GDF-9、BMP-15的mRNA表达水平有明显下降,而颗粒细胞源性自分泌、旁分泌因子SCF、bFGF的mRNA表达水平则无变化。结论: 慢性应激可以影响小鼠的动情周期,影响小鼠卵泡的发育及排卵。外源性补充促性腺激素后,慢性应激组排卵数明显减少,HE染色结果显示:慢性应激增加了卵泡的闭锁,干扰了卵泡的发育和排卵,即使外源性给予促性腺激素仍不能使早期卵泡发育和排卵恢复正常,提示非促性腺激素类因素参与慢性应激干扰卵泡发育和排卵。RT-PCR结果显示,慢性应激后,相比正常对照组,自分泌、旁分泌因子GDF-9、BMP-15表达水平有明显降低(P0.05),SCF、bFGF表达无明显变化。提示慢性应激通过下调卵源性的局部旁分泌因子GDF-9、BMP-15参与影响卵泡的发育。
[Abstract]:Aim: to observe the effects of chronic stress on follicular development and estrus cycle and on the non-gonadotropin dependence of follicular development. Four representative ovarian paracrine factors of GDF-9,BMP-15,bFGF,SCF and autocrine factors were selected to observe whether chronic stress affects the expression of local ovarian regulatory factors which regulate follicular development. Methods: (1) chronic stress model was established in mice. The effects of chronic stress on follicle development were observed by preparing ovarian HE sections. (2) from the first day of chronic stress to the day of death, the vaginal secretions of mice were taken as smears. The estrus cycle was observed by RIA staining. (3) supplementation of gonadotropin, (4) relative quantitative real-time PCR method was used to measure the mRNA level of paracrine and autocrine factors of GDF-9,BMP-15,bFGF,SCF. The results were as follows: (1) chronic stress caused the disorder of estrous cycle in mice. The interval was prolonged. (2) chronic stress affected follicle development and ovulation. (3) exogenous supplementation of gonadotropin could not completely restore follicle development and ovulation to normal. (4) oocyte autocrine, The mRNA expression of paracrine factor GDF-9,BMP-15 was significantly decreased, while that of granulosa cell derived autocrine was not changed, while that of paracrine factor SCF,bFGF was not changed. Conclusion: chronic stress can affect the oestrus cycle, follicle development and ovulation of mice. After exogenous supplementation of gonadotropin, the number of ovulation decreased significantly in chronic stress group. The results showed that chronic stress increased follicle atresia and interfered with follicle development and ovulation. Even exogenous administration of gonadotropin could not make early follicle development and ovulation return to normal, suggesting that non-gonadotropins participate in chronic stress to interfere follicle development and ovulation. RT-PCR results showed that, after chronic stress, compared with normal control group, non-gonadotropins involved in follicular development and ovulation. The expression of autocrine and paracrine factor GDF-9,BMP-15 was significantly decreased (P0.05). The results suggest that chronic stress affects follicular development by down-regulating the local paracrine factor (GDF-9,BMP-15).
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R33

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