妊娠期奶山羊黄体中褪黑素膜受体MT1和MT2的分布和表达
发布时间：2018-07-15 18:33
【摘要】：褪黑素(melatonin)是一种由哺乳动物松果体分泌的神经内分泌胺类激素,通过其高亲和性膜受体(membrane receptors,MT)的介导参与调控动物生理节律、季节性繁殖和卵巢功能等生理过程。人和其他哺乳类动物的高亲和性膜受体分为Mel1a(MT1)、Mel1b(MT2)两种亚型。研究发现MT1和MT2在动物下丘脑、视网膜、脑垂体、睾丸、卵巢、胃肠道和乳腺等均有表达。迄今为止,MT1和MT2在妊娠期奶山羊黄体的分布和表达尚未见报道。本试验以妊娠期关中奶山羊为研究对象,采用免疫组化、QRT-PCR和Western blot等方法对妊娠期不同阶段奶山羊黄体中MT1和MT2的分布和表达进行了探索。研究取得如下结果。1.在妊娠期不同阶段奶山羊卵巢中都可检测到MT1和MT2的表达,妊娠前期和中期的表达水平(mRNA和蛋白水平)显著高于妊娠后期(P0.01)。2.免疫组化结果表明,MT1和MT2在妊娠期不同阶段黄体组织中都有分布,主要定位于黄体细胞。MT1和MT2在妊娠前期和妊娠中期黄体细胞中免疫阳性反应较强,尤其是在妊娠中期,几乎所有的黄体细胞都为强阳性反应。妊娠后期黄体细胞MT1和MT2的免疫阳性反应较弱。3.qRT-PCR和Western blot检测结果表明,在妊娠期不同阶段奶山羊黄体组织中MT1和MT2表达(mRNA和蛋白水平)规律与在卵巢中表达相似,妊娠前期和妊娠中期显著高于妊娠后期(P0.01)。4.Melatonin处理体外培养黄体细胞,结果发现melatonin剂量依赖性的提高黄体细胞中MT1和MT2的表达(mRNA和蛋白水平);另外,CCK-8检测细胞活性结果表明,200和2000 pg/mL浓度的melatonin处理黄体细胞36h和48h后细胞活性显著增加(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).
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S827.3
本文编号：2125008
[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).
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S827.3
【参考文献】
相关期刊论文 前5条
1 孙中安,刘志民,黄超;人胚胎甲状腺褪黑素受体mRNA的检测[J];第二军医大学学报;2000年11期
2 赵瑛,邵福源,何淑芬,彭树勋;人胚胎中枢神经系统褪黑素受体的鉴定及生物学特性[J];第二军医大学学报;2001年01期
3 陆祖谦,刘志民,何金,刘会敏,赵瑛,彭树勋;人胚胎肾上腺皮质褪黑素受体亚型蛋白的分布[J];第二军医大学学报;2001年01期
4 王国权,刘冬萍,邵福源,赵瑛,刘志民;褪黑素受体在人胎脑中的分布[J];江西医学院学报;2003年03期
5 周爱民,袁育康,范桂香,王军阳,任会勋;褪黑素的免疫调节作用[J];西安医科大学学报;2001年05期
，本文编号：2125008
本文链接：https://www.wllwen.com/yixuelunwen/dongwuyixue/2125008.html
