INHA基因干扰对马关无角山羊卵巢颗粒细胞的影响  

Effect of Interference of INHA Gene on Ovarian Granular Cells of Maguan Polled Goats

作　　者：富国文[1] 王绍卿[1] 滕晓红[1] 达永仙[1] 樊月圆[1] 

机构地区：[1]云南农业大学动物科学技术学院,云南昆明650201

出　　处：《家畜生态学报》2016年第8期12-15,49,共5页Journal of Domestic Animal Ecology

基　　金：云南省科技计划面上项目(2012FB147)

摘　　要：为了揭示INHA基因在马关无角山羊卵泡发育过程中的作用,为山羊多胎性状的研究提供理论依据,以高繁殖力地方品种--马关无角山羊的卵巢颗粒细胞为研究对象,采用siRNA技术,对卵巢颗粒细胞所分泌的糖蛋白激素基因INHA进行了干扰,Western blot法检验了干扰后INHA蛋白的表达量,Real-time PCR法检测了干扰后凋亡家族主要基因、主要生长因子及激素受体的mRNA表达量变化。Western blot结果显示,干扰48h后的INHA蛋白的表达量极显著低于正常组和阴性对照组(negative control,NC)(P<0.01);Real-time PCR结果显示,凋亡促进基因Bax在转染后,表达量极显著低于正常组和NC组(P<0.01),凋亡抑制基因bcl-2和bcl-xl基因在转染后,表达量极显著高于正常组和NC组(P<0.01);主要生长因子IGF-1和IGF-2基因在转染后,mRNA的表达量明显减少(P<0.01),其中IGF-1的表达量降低最为明显;激素受体FSHR和LHR基因mRNA的表达量极显著低于正常组和NC组(P<0.01)。由此推测,INHA基因可能是通过影响卵巢颗粒细胞凋亡/增殖,进而影响了卵泡的发育。In order to reveal the role of INHAgene in follicle development of Maguan polled goats,and provide a theoretical basis for the research of goat multiplets traits,this study took the ovarian granulosa cells of Maguan polled goat-a local variety-as the research object,using siRNA interference glycoprotein hormone gene INHA which is secreted by ovarian granulosa cells,the INHA protein expression after interference was tested by Western-blotting,main apoptosis regulation genes was tested by Real-time PCR,changes of growth factors and hormone receptor mRNA expression quantity were also detected.Western-blotting results show that the INHA protein expression was significantly lower than normal group and NC group（P〈0.01）after interference for 48 h.Real-time PCR results show that apoptosis gene Bax expression was significantly lower than the other two groups（P〈0.01）.After transfection,expressing quantity of apoptosis suppressor gene BCL-2and BCL-xl was significantly higher than that of the others（P〈0.01）,the gene mRNA expression quantity of major growth factor IGF-1and IGF-2significantly reduced（P〈0.01）,especially IGF-1.mRNA expression of FSHRand LHRgene significantly were lower than normal group and NC group（P〈0.01）.So it is speculated that INHAgene may affect the development of follicles by influencing the apoptosis/proliferation of ovarian granulosa cell.

关 键 词：INHA基因 卵巢颗粒细胞 SIRNA技术 马关无角山羊 

分 类 号：S811.5[农业科学—畜牧学]