本文選題：�；撬� + 下丘腦-垂體-卵巢軸　； 參考：《沈陽農(nóng)業(yè)大學》2015年博士論文

【摘要】：牛磺酸作為人和動物的條件性必需氨基酸,是雌性動物生殖腺和生殖細胞中含量最為豐富的游離氨基酸之一,已有研究證明�；撬崤c卵巢組織發(fā)育成熟及雌激素合成分泌密切相關(guān),但其具體機制尚不清楚。本研究以雌性大鼠為試驗動物,從mRNA和蛋白水平檢測了�；撬嵘锖铣申P(guān)鍵酶—半胱氨酸雙加氧酶(CDO)、半胱亞磺酸脫羧酶(CSD)及�；撬徂D(zhuǎn)運體(TAUT)在下丘腦-垂體-卵巢軸中的表達；通過牛磺酸飼喂試驗,檢測了�；撬釋Σ煌l(fā)情周期大鼠生殖激素分泌的影響；采用體外細胞培養(yǎng)技術(shù),檢測了�；撬釋w外培養(yǎng)大鼠下丘腦細胞、垂體細胞、卵巢細胞激素分泌及相關(guān)因子表達的影響；通過體外培養(yǎng)卵泡和卵母細胞檢測了牛磺酸對卵泡發(fā)育、卵母細胞成熟、受精及卵裂的影響,探討了牛磺酸對雌性生殖機能作用的影響及其可能機制,以期為�；撬嵩谂约按菩詣游锷持械膽锰峁├碚撘罁�(jù)。研究結(jié)果如下：1.成功克隆出了大鼠下丘腦-垂體-卵巢軸上CSD基因的編碼區(qū)序列,下丘腦CDO基因的編碼區(qū)序列,卵巢CDO基因的全長序列及下丘腦-垂體-卵巢軸上TAUT基因的全長序列,且與肝臟中基因的核苷酸序列、氨基酸序列均有較高同源率(96%以上)；下丘腦-垂體-卵巢軸上也存在CSD、CDOTAUT蛋白表達；CDO、CSDTAUT主要分布于卵泡內(nèi)膜間質(zhì)細胞層。結(jié)果表明：下丘腦、垂體和卵巢能通過CSD通路合成牛磺酸,并具有轉(zhuǎn)運外源�；撬崛氚哪芰Α�2.�；撬釋Σ煌l(fā)情周期大鼠促性腺激素釋放激素(GnRH)、泌乳素(PRL)、促黃體生成素(LH)、雌二醇(E2)和孕酮(P)的分泌均有一定的刺激作用,但對促卵泡刺激素(FSH)分泌的影響不明顯。結(jié)果表明：�；撬峥赡芡ㄟ^作用于下丘腦-垂體-卵巢軸來調(diào)控雌性大鼠的生殖機能。3.�；撬崮艽龠M體外培養(yǎng)大鼠下丘腦細胞GnRH、腺垂體細胞PRL和LH、卵巢內(nèi)膜細胞T、顆粒細胞E2和P的分泌；提高內(nèi)膜細胞中促黃體生成素受體(LHR)、3β-羥基類固醇脫氫酶(3p-HSD)和17p-羥基類固醇脫氫酶(17P-HSD) mRNA的表達,增加顆粒細胞中促卵泡刺激素受體(FSHR)和CYP-19A-1 mRNA的表達。結(jié)果表明：牛磺酸可通過增加卵巢中FSHR和LHR的數(shù)量,促進性激素合成關(guān)鍵酶的表達來促進雌激素的合成和分泌。4.�；撬峥纱龠M離體培養(yǎng)卵泡直徑增長和E2的分泌；�；撬釋﹄x體培養(yǎng)卵母細胞MⅡ期形成、受精及卵裂具有促進作用。結(jié)果表明：�；撬崮艽龠M卵泡發(fā)育和卵母細胞成熟、受精及卵裂率。本文試驗結(jié)果表明,大鼠下丘腦-垂體-卵巢軸均可生物合成牛磺酸,而且具有將外源�；撬徂D(zhuǎn)運入胞的能力；�；撬峥赏ㄟ^增強下丘腦-垂體-卵巢軸的功能來調(diào)控雌性生殖機能。
[Abstract]:Taurine is one of the most abundant free amino acids in the gonads and germ cells of female animals. Studies have shown that taurine is closely related to ovarian tissue maturation and estrogen synthesis and secretion, but its mechanism is not clear. In this study, the expression of cysteine dioxygenase (cysteine dioxygenase), cysteine decarboxylase (cysteine decarboxylase) and taurine transporter (taurine) in hypothalamus-pituitary-ovary axis was detected from mRNA and protein levels in female rats. The effects of taurine on the secretion of reproductive hormones in rats with different estrous cycles were tested by taurine feeding test, and the effects of taurine on rat hypothalamic and pituitary cells in vitro were detected by cell culture technique in vitro. The effects of taurine on follicle development, oocyte maturation, fertilization and cleavage were examined by in vitro culture of follicles and oocytes. The effect of taurine on female reproductive function and its possible mechanism were discussed in order to provide theoretical basis for the application of taurine in female and female animal reproduction. The results are as follows: 1. The coding region sequence of CSD gene on hypothalamus-pituitary-ovary axis, the coding region sequence of CDO gene in hypothalamus, the full-length sequence of CDO gene in ovary and the full-length sequence of TAUT gene on hypothalamus-pituitary-ovary axis were cloned successfully. Moreover, the nucleotide sequence and amino acid sequence of the gene in the liver had a high homology rate of more than 96%, and the expression of CSD- CDOTAUT protein on the hypothalamus-pituitary-ovary axis was mainly distributed in the interstitial layer of the follicular intima. The results showed that the hypothalamus, pituitary and ovary could synthesize taurine via CSD pathway and have the ability of transporting exogenous taurine into cells. Taurine stimulated the secretion of gonadotropin releasing hormone (GnRH), prolactin (PRL), luteinizing hormone (LH), estradiol (E 2) and progesterone (P) in rats with different estrus cycle, but had no obvious effect on follicle stimulating hormone (FSH) secretion. The results showed that taurine may regulate the reproductive function of female rats by acting on hypothalamus-pituitary-ovarian axis. Taurine could promote the secretion of GnRH in rat hypothalamic cells, PRL and LHin adenohypophysis cells, T, E 2 and P in granulosa cells. To increase the expression of luteinizing hormone receptor (LHR3 尾 -hydroxysteroid dehydrogenase 3p-HSD3) and 17p- hydroxysteroid dehydrogenase (17P-HSD) mRNA, and to increase the expression of follicle-stimulating hormone receptor (FSHR) and CYP-19A-1 mRNA in granulosa cells. The results showed that taurine could promote the synthesis and secretion of estrogen by increasing the number of FSHR and LHR in ovary and promoting the expression of key enzymes of sex hormone synthesis. Taurine could increase the diameter of follicle and secretion of E2 in vitro, and taurine could promote the formation of M 鈪，

本文編號：1864032