【摘要】：褪黑素(melatonin)是一種由哺乳動(dòng)物松果體分泌的神經(jīng)內(nèi)分泌胺類激素,通過(guò)其高親和性膜受體(membrane receptors,MT)的介導(dǎo)參與調(diào)控動(dòng)物生理節(jié)律、季節(jié)性繁殖和卵巢功能等生理過(guò)程。人和其他哺乳類動(dòng)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(dòng)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(dá)。迄今為止,MT1和MT2在妊娠期奶山羊黃體的分布和表達(dá)尚未見(jiàn)報(bào)道。本試驗(yàn)以妊娠期關(guān)中奶山羊?yàn)檠芯繉?duì)象,采用免疫組化、QRT-PCR和Western blot等方法對(duì)妊娠期不同階段奶山羊黃體中MT1和MT2的分布和表達(dá)進(jìn)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(cè)到MT1和MT2的表達(dá),妊娠前期和中期的表達(dá)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(xì)胞。MT1和MT2在妊娠前期和妊娠中期黃體細(xì)胞中免疫陽(yáng)性反應(yīng)較強(qiáng),尤其是在妊娠中期,幾乎所有的黃體細(xì)胞都為強(qiáng)陽(yáng)性反應(yīng)。妊娠后期黃體細(xì)胞MT1和MT2的免疫陽(yáng)性反應(yīng)較弱。3.qRT-PCR和Western blot檢測(cè)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(dá)(mRNA和蛋白水平)規(guī)律與在卵巢中表達(dá)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(xì)胞,結(jié)果發(fā)現(xiàn)melatonin劑量依賴性的提高黃體細(xì)胞中MT1和MT2的表達(dá)(mRNA和蛋白水平);另外,CCK-8檢測(cè)細(xì)胞活性結(jié)果表明,200和2000 pg/mL濃度的melatonin處理黃體細(xì)胞36h和48h后細(xì)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(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S827.3

【參考文獻(xiàn)】

相關(guān)期刊論文 前5條

1 孫中安,劉志民,黃超;人胚胎甲狀腺褪黑素受體mRNA的檢測(cè)[J];第二軍醫(yī)大學(xué)學(xué)報(bào);2000年11期

2 趙瑛,邵福源,何淑芬,彭樹(shù)勛;人胚胎中樞神經(jīng)系統(tǒng)褪黑素受體的鑒定及生物學(xué)特性[J];第二軍醫(yī)大學(xué)學(xué)報(bào);2001年01期

3 陸祖謙,劉志民,何金,劉會(huì)敏,趙瑛,彭樹(shù)勛;人胚胎腎上腺皮質(zhì)褪黑素受體亞型蛋白的分布[J];第二軍醫(yī)大學(xué)學(xué)報(bào);2001年01期

4 王國(guó)權(quán),劉冬萍,邵福源,趙瑛,劉志民;褪黑素受體在人胎腦中的分布[J];江西醫(yī)學(xué)院學(xué)報(bào);2003年03期

5 周愛(ài)民,袁育康,范桂香,王軍陽(yáng),任會(huì)勛;褪黑素的免疫調(diào)節(jié)作用[J];西安醫(yī)科大學(xué)學(xué)報(bào);2001年05期

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本文編號(hào)：2125008