本文關鍵詞： 著床前束縛應激 糖皮質激素 后代焦慮 卵母細胞發(fā)育能力 胚胎培養(yǎng) 凋亡 Fas/FasL 小鼠　出處：《山東農業(yè)大學》2016年博士論文　論文類型：學位論文

【摘要】：PART I:研究表明應激可以導致糖皮質激素過量分泌并損傷卵母細胞發(fā)育能力。我們實驗室前期工作發(fā)現(xiàn)注射糖皮質激素(皮質醇)同樣導致小鼠卵母細胞發(fā)育能力下降,但體外成熟過程中添加應激生理濃度的糖皮質激素并不影響卵母細胞的成熟和胚胎的發(fā)育。本實驗主要通過檢測注射皮質醇后小鼠卵母細胞的發(fā)育能力,卵巢細胞凋亡情況,Fas/Fas L信號通路的激活來探索糖皮質激素損傷卵母細胞發(fā)育能力的具體機制。結果顯示注射糖皮質激素(皮質醇)顯著降低雌性小鼠(a)卵母細胞發(fā)育能力,(b)血清和卵巢中E2/P4的比例,(c)壁顆粒細胞中胰島素樣生長因子1(IGF1)、腦源性營養(yǎng)因子(BDNF)及糖皮質激素受體(GR)的表達。同時顯著升高(a)血清和卵巢中皮質醇的含量,(b)壁顆粒細胞和卵丘細胞的凋亡比例,(c)體外成熟培養(yǎng)過程中卵丘細胞分泌的Fas L,(d)壁顆粒細胞、卵丘細胞和卵母細胞中Fas的表達。當給Fas L基因突變小鼠(gld小鼠)注射皮質醇后,發(fā)現(xiàn)其卵母細胞發(fā)育能力的削弱程度、壁顆粒細胞和卵丘細胞的凋亡比例與野生型相比都有很大幅度的緩解。由此可以得出以下結論:糖皮質激素是通過激活Fas系統(tǒng)導致卵巢細胞凋亡進而損傷卵母細胞發(fā)育潛能。到目前為止,本實驗是首次揭示糖皮質激素降低卵母細胞發(fā)育能力的機制,這將有助于進一步理解糖皮質激素在應激相關疾病中作用。PART II:人類流行病學研究表明產(chǎn)前應激可能會損傷情感發(fā)育,增加后代患抑郁癥、焦慮癥、精神分裂癥、自閉癥的機率。但是產(chǎn)前應激影響后代行為和神經(jīng)內分泌系統(tǒng)的機制尚不十分清楚。另外,大多數(shù)這方面的研究都集中于妊娠晚期。關于著床前應激對后代影響的研究很少,而有關這方面的體外試驗更是缺乏。本實驗主要研究著床前心理應激是否影響后代焦慮行為,并且運用體內和體外外兩種模型研究后代行為的改變是否是通過應激增加糖皮質激素的分泌引起的。結果表明著床前應激和體外培養(yǎng)胚胎過程中添加糖皮質激素(皮質酮)都可以增加后代焦慮行為,導致后代海馬區(qū)糖皮質激素受體(GR)和腦源性營養(yǎng)因子(BDNF)的下調,并促進糖皮質激素的分泌。而且體外培養(yǎng)過程中添加皮質酮會下調胚胎內糖皮質激素受體的表達。由此可以得出以下結論:著床前心理應激是通過促進妊娠母鼠糖皮質激素的分泌,進而引起胚胎內糖皮質激素受體表達下降,最終導致后代焦慮水平上升。本研究是首次報道著床前應激和體外培養(yǎng)胚胎時添加糖皮質激素會增加后代的焦慮行為,這些實驗結果可以幫助我們進一步理解產(chǎn)前應激影響后代行為的具體機制。
[Abstract]:PART I: studies have shown that stress can cause hypersecretion of glucocorticoids and damage oocyte development. Our laboratory work found that injecting glucocorticoid (cortisol). The development ability of mouse oocytes was also decreased. However, glucocorticoids added to stress physiological concentration during in vitro maturation did not affect the maturation of oocytes and the development of embryos. Apoptosis of ovarian cells. Activation of Fas/Fas L signaling pathway to explore the specific mechanism of glucocorticoid injury on oocyte development. Oocyte development. The ratio of E _ 2 / P _ 4 in serum and ovary was found in parietal granulosa cells, and insulin-like growth factor-1 (IGF1) was found in parietal granulosa cells. The expression of brain-derived nutrition factor (BDNF) and glucocorticoid receptor (GRG) increased significantly (P < 0.05). Meanwhile, the content of cortisol in serum and ovary was significantly higher than that in parietal granulosa cells and cumulus cells. (C) parietal granulosa cells secreted by cumulus cells during maturation in vitro. The expression of Fas in cumulus cells and oocytes. After injecting cortisol into Fas L mutant mice (GLD mice), it was found that the oocyte development ability was weakened. The percentage of apoptosis in parietal granulosa cells and cumulus cells was significantly alleviated compared with wild-type cells. Glucocorticoid induces ovarian cell apoptosis by activating the Fas system and thus impairs oocyte developmental potential. This experiment is the first to reveal the mechanism of glucocorticoid decreasing oocyte development ability. This will help to further understand the role of glucocorticoids in stress-related diseases .PART II: human epidemiological studies have shown that prenatal stress may damage emotional development and increase depression in offspring. The risk of anxiety, schizophrenia, and autism. But the mechanisms by which prenatal stress affects offspring's behavior and neuroendocrine system are not well understood. Most of these studies have focused on late pregnancy, and little research has been done on the effects of pre-implantation stress on offspring. However, there is a lack of in vitro experiments in this field. This study mainly studied whether pre-implantation psychological stress affects the anxiety behavior of offspring. Two models in vivo and in vitro were used to study whether the behavior change of offspring was caused by stress increasing the secretion of glucocorticoid. The results showed that preimplantation stress and in vitro embryo culture were supplemented with glucocorticoid. (. Corticosterone) can increase anxiety behavior in offspring. This results in down-regulation of glucocorticoid receptor (GRG) and brain-derived nutrition factor (BDNF) in hippocampal area of offspring. And to promote the secretion of glucocorticoids. Moreover, adding corticosterone in vitro can down-regulate the expression of glucocorticoid receptors in embryos. Preimplantation psychological stress promotes the secretion of glucocorticoid in pregnant rats. In turn, the expression of glucocorticoid receptor in embryos decreased. This is the first study to report that pre-implantation stress and glucocorticoid supplementation in vitro embryo culture can increase anxiety behavior in offspring. These results can help us to further understand the specific mechanism of prenatal stress affecting offspring behavior.
【學位授予單位】：山東農業(yè)大學
【學位級別】：博士
【學位授予年份】：2016
【分類號】：Q492

【參考文獻】

相關期刊論文 前2條

1 ;Bioactive compounds from Paecilomyces tenuipes regulating the function of the hypothalamo-hypophyseal system axis in chronic unpredictable stress rats[J];Chinese Medical Journal;2007年12期

2 ;Apoptosis in Granulosa cells during follicular atresia: relationship with steroids and insulin-like growth factors[J];Cell Research;2004年04期

，

本文編號：1461677