【摘要】：褪黑素(melatonin)是一種由哺乳動物松果體分泌的神經(jīng)內(nèi)分泌胺類激素,通過其高親和性膜受體(membrane receptors,MT)的介導(dǎo)參與調(diào)控動物生理節(jié)律、季節(jié)性繁殖和卵巢功能等生理過程。人和其他哺乳類動物的高親和性膜受體分為Mel1a(MT1)、Mel1b(MT2)兩種亞型。研究發(fā)現(xiàn)MT1和MT2在動物下丘腦、視網(wǎng)膜、腦垂體、睪丸、卵巢、胃腸道和乳腺等均有表達。迄今為止,MT1和MT2在妊娠期奶山羊黃體的分布和表達尚未見報道。本試驗以妊娠期關(guān)中奶山羊為研究對象,采用免疫組化、QRT-PCR和Western blot等方法對妊娠期不同階段奶山羊黃體中MT1和MT2的分布和表達進行了探索。研究取得如下結(jié)果。1.在妊娠期不同階段奶山羊卵巢中都可檢測到MT1和MT2的表達,妊娠前期和中期的表達水平(mRNA和蛋白水平)顯著高于妊娠后期(P0.01)。2.免疫組化結(jié)果表明,MT1和MT2在妊娠期不同階段黃體組織中都有分布,主要定位于黃體細胞。MT1和MT2在妊娠前期和妊娠中期黃體細胞中免疫陽性反應(yīng)較強,尤其是在妊娠中期,幾乎所有的黃體細胞都為強陽性反應(yīng)。妊娠后期黃體細胞MT1和MT2的免疫陽性反應(yīng)較弱。3.qRT-PCR和Western blot檢測結(jié)果表明,在妊娠期不同階段奶山羊黃體組織中MT1和MT2表達(mRNA和蛋白水平)規(guī)律與在卵巢中表達相似,妊娠前期和妊娠中期顯著高于妊娠后期(P0.01)。4.Melatonin處理體外培養(yǎng)黃體細胞,結(jié)果發(fā)現(xiàn)melatonin劑量依賴性的提高黃體細胞中MT1和MT2的表達(mRNA和蛋白水平);另外,CCK-8檢測細胞活性結(jié)果表明,200和2000 pg/mL濃度的melatonin處理黃體細胞36h和48h后細胞活性顯著增加(P0.01)。
[Abstract]:Melatonin (melatonin) (melatonin (melatonin) is a neuroendocrine amine hormone secreted by mammalian pineal gland. It is involved in the regulation of physiological rhythm, seasonal reproduction and ovarian function through the mediation of high affinity membrane receptor (membrane receptor). The high affinity membrane receptors of human and other mammals are classified into two subtypes: Mel1a (MT1) and Mel1b (MT2). It was found that MT1 and MT2 were expressed in hypothalamus, retina, pituitary, testis, ovary, gastrointestinal tract and mammary gland. Up to now, the distribution and expression of MT1 and MT2 in corpus luteum of dairy goats during pregnancy have not been reported. In this study, the distribution and expression of MT1 and MT2 in corpus luteum of Guanzhong dairy goats during pregnancy were studied by immunohistochemical QRT-PCR and Western blot. The results of the study are as follows. The expression of MT1 and MT2 was detected in the ovaries of dairy goats at different stages of pregnancy. The expression levels of MT1 and MT2 (mRNA and protein) in the first and second trimester of pregnancy were significantly higher than those in the second trimester (P0.01). The immunohistochemical results showed that both MT1 and MT2 were distributed in luteal tissues in different stages of pregnancy, mainly located in luteal cells. MT1 and MT2 were strongly immunoreactive in luteal cells in prepregnancy and second trimester of pregnancy, especially in the second trimester of pregnancy. Almost all luteal cells were strongly positive. The expression patterns of MT1 and MT2 (mRNA and protein levels) in luteal tissues of dairy goats at different stages of pregnancy were similar to those in ovaries. The levels of luteal cells in vitro treated with Melatonin were significantly higher than those in the second trimester of pregnancy (P0.01) and in the second trimester of pregnancy. The results showed that the expression of MT1 and MT2 in luteal cells was increased in a dose-dependent manner by melatonin, and the activity of luteal cells was detected by CCK-8 assay in a dose-dependent manner. The results showed that the cellular activity of luteal cells treated with melatonin at the concentration of 2000 PG / mL for 36 h and 48 h was significantly increased (P0.01).
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S827.3

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本文編號：2125008