SMAD7对山羊卵泡颗粒细胞增殖、凋亡的影响  

Effects of SMAD7on Proliferation and Apoptosis of Goat Follicular Granulosa Cells

作　　者：李碧筠 黄思艺 王钰锟 王磊[1] 何莉娜 唐雪 徐德军 赵中权[1] LI Bijun;HUANG Siyi;WANG Yukun;WANG Lei;HE Lina;TANG Xue;XU Dejun;ZHAO Zhongquan(College of Animal Science and Technology,Southwest University,Chongqing400715,China)

机构地区：[1]西南大学动物科学技术学院,重庆400715

出　　处：《畜牧兽医学报》2022年第8期2548-2557,共10页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：重庆市技术创新与应用发展专项(cstc2021jscx-gksbX0008);中央高校基本科研业务费(XDJK2020B013)。

摘　　要：旨在探究SMAD7对山羊卵泡颗粒细胞增殖和凋亡的影响。本试验收集3~4月龄大足黑山羊母羊的卵泡颗粒细胞,通过过表达或siRNA干扰、ELISA、qRT-PCR、Western blot及流式细胞术等技术与方法探究SMAD7对颗粒细胞增殖、凋亡及类固醇激素分泌的影响。结果发现,SMAD7过表达显著下调颗粒细胞增殖活力并促进细胞凋亡,抑制PCNA表达(P<0.05),下调BCL2/BAX的比值(P<0.01);同时,SMAD7干扰显著上调颗粒细胞增殖活力,显著上调PCNA表达(P<0.05)与BCL2/BAX表达量比值(P<0.05)。SMAD7过表达极显著上调颗粒细胞的孕酮分泌,下调雌二醇表达水平(P<0.01);同时SMAD7干扰极显著下调孕酮分泌,上调雌二醇分泌(P<0.01)。进一步研究发现,SMAD7过表达显著抑制SMAD2、SMAD3的mRNA和蛋白表达(P<0.05);SMAD7干扰则显著促进SMAD2、SMAD3的mRNA和蛋白表达(P<0.05)。结果表明,SMAD7抑制山羊卵泡颗粒细胞的增殖和雌二醇分泌,促进凋亡和孕酮的合成,并且抑制SMAD2、SMAD3的表达,进而调节卵泡的发育与闭锁。The purpose of this study was to explore the effects of SMAD7on the proliferation and apoptosis of follicular granulosa cells in goat.In this study,follicular granulosa cells were collected from 3-to 4-month-old Dazu black goats.The overexpression,siRNA interference,ELISA,qRT-PCR,Western blot and flow cytometry were used to investigate the effects of SMAD7on proliferation,apoptosis and steroid secretion of granulosa cells.The results showed that overexpression of SMAD7significantly reduced the proliferation activity and promoted the apoptosis of granulosa cells,inhibited the expression of PCNA(P<0.05)and decreased the ratio of BCL2/BAX(P<0.01).Oppositely,SMAD7interference significantly promoted the proliferation activity of granulosa cells,significantly up-regulated the expression of PCNAand the ratio of BCL2/BAX(P<0.05).SMAD7overexpression significantly increased progesterone secretion of granulated cells,and reduced estradiol expression levels(P<0.01).Meanwhile,SMAD7interference significantly down-regulated the secretion of progesterone and up-regulated the secretion of estradiol(P<0.01).Further studies showed that SMAD7overexpression significantly inhibited SMAD2,SMAD3mRNA and protein expression(P<0.05),but did not affect SMAD4ex pression.SMAD7interference significantly promoted the mRNA and protein levels of SMAD2,SMAD3(P<0.05).The results suggested that SMAD7inhibits the proliferation and estradiol secretion,promotes apoptosis and progesterone synthesis,and inhibits the expression of SMAD2,SMAD3in follicular granulosa cells,thereby regulating the development and atresia of follicles.

关 键 词：SMAD7 山羊 颗粒细胞 增殖 凋亡 

分 类 号：S827.3[农业科学—畜牧学]