【摘要】： 目的: 觀察慢性應激對卵泡發(fā)育和動情周期的影響及其對卵泡發(fā)育的非促性腺激素依賴的影響。選取GDF-9、BMP-15、bFGF、SCF四個代表性的卵巢旁分泌、自分泌因子行RT-PCR,觀察慢性應激是否影響這些調控卵泡發(fā)育的卵巢局部調控因子的表達。 方法: (1)通過建立小鼠慢性應激模型,制備卵巢HE切片觀察慢性應激對卵泡發(fā)育的影響。 (2)從慢性應激的第一天開始(至處死日止)取小鼠陰道分泌物,作涂片,行瑞氏染色觀察其所處動情周期。 (3)通過外源性補充促性腺激素,觀察其對應激后卵泡發(fā)育的恢復作用。 (4)采用相對定量real-time PCR方法測定GDF-9、BMP-15、bFGF、SCF這幾個代表性的卵巢旁分泌、自分泌因子的mRNA水平。 實驗結果: (1)慢性應激使小鼠的動情周期紊亂、間期延長。 (2)慢性應激影響卵泡的發(fā)育及排卵的發(fā)生。 (3)外源性補充促性腺激素不能使卵泡發(fā)育和排卵完全恢復正常。 (4)卵源性自分泌、旁分泌因子GDF-9、BMP-15的mRNA表達水平有明顯下降,而顆粒細胞源性自分泌、旁分泌因子SCF、bFGF的mRNA表達水平則無變化。 結論: 慢性應激可以影響小鼠的動情周期,影響小鼠卵泡的發(fā)育及排卵。外源性補充促性腺激素后,慢性應激組排卵數(shù)明顯減少,HE染色結果顯示:慢性應激增加了卵泡的閉鎖,干擾了卵泡的發(fā)育和排卵,即使外源性給予促性腺激素仍不能使早期卵泡發(fā)育和排卵恢復正常,提示非促性腺激素類因素參與慢性應激干擾卵泡發(fā)育和排卵。RT-PCR結果顯示,慢性應激后,相比正常對照組,自分泌、旁分泌因子GDF-9、BMP-15表達水平有明顯降低(P0.05),SCF、bFGF表達無明顯變化。提示慢性應激通過下調卵源性的局部旁分泌因子GDF-9、BMP-15參與影響卵泡的發(fā)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).
【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R33

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