摘要
为探究miRNA-148a-3p在山羊卵巢颗粒细胞中的作用,通过过表达或抑制miRNA-148a-3p、qRT-PCR、Western blot、流式细胞术、ELISA等方法,探究miR-148a-3p对卵巢颗粒细胞增殖、凋亡、自噬及类固醇激素分泌的影响。结果发现,过表达miR-148a-3p显著促进卵巢颗粒细胞增殖和PCNA表达,抑制miR-148a-3p显著抑制卵巢颗粒细胞增殖和PCNA表达。过表达miR-148a-3p抑制卵巢颗粒细胞凋亡,极显著下调细胞凋亡率,显著上调Bcl-2/Bax的mRNA和蛋白比值,抑制miR-148a-3p促进卵巢颗粒细胞凋亡。过表达miR-148a-3p显著促进卵巢颗粒细胞自噬,上调LC3-Ⅱ/LC3-Ⅰ比值,抑制miR-148a-3p上调p62表达。过表达miR-148a-3p显著促进卵巢颗粒细胞孕酮分泌,并上调3β-HSD、StAR、CYP11A1的mRNA和蛋白表达。虽然抑制miR-148a-3p下调孕酮水平,但对3β-HSD、StAR、CYP11A1及CYP19A1的mRNA和蛋白表达无显著性影响。结果表明,miR-148a-3p可促进卵巢颗粒细胞增殖、抑制细胞凋亡、诱导自噬并刺激孕酮的合成。
To investigate the role of miRNA-148a-3p in goat ovarian granulosa cells,in this study,overexpression or inhibition of miRNA-148a-3p,qRT-PCR,Western blot,flow cytometry,ELISA and other methods were used to explore the effects of miR-148a-3p on proliferation,apoptosis,autophagy and steroid hormones secretion of ovarian granulosa cells.The results showed that overexpression of miR-148a-3p significantly promoted the proliferation and PCNA expression of ovarian granulosa cells,and inhibition of miR-148a-3p significantly inhibited the proliferation and PCNA expression of ovarian granulosa cells.Overexpression of miR-148a-3p inhibited apoptosis of ovarian granulosa cells,significantly down-regulated apoptosis rate,significantly up-regulated mRNA and protein ratio of Bcl-2/Bax,and inhibited miR-148a-3p promoted apoptosis of ovarian granulosa cells.Overexpression of miR-148a-3p significantly promoted the autophagy of ovarian granulosia cells,up-regulated LC3-Ⅱ/LC3-Ⅰratio,and inhibited the up-regulated p62expression of miR-148a-3p.Overexpression of miR-148a-3p significantly promoted progesterone secretion of ovarian granulosa cells,and up-regulated mRNA and protein expressions of 3β-HSD,StAR and CYP11A1.Although inhibition of miR-148a-3p down-regulated progesterone level,mRNA and protein expressions of 3β-HSD,StAR,CYP11A1and CYP19A1 were not affected.The results showed that miR-148a-3p promoted the proliferation of ovarian granulosa cells,inhibited apoptosis,induced autophagy,and stimulated progesterone synthesis.
作者
王钰锟
李碧筠
张义语
丁文飞
王磊
唐雪
宋帅飞
姚慧
黄德利
徐德军
赵中权
WANG Yukun;LI Bijun;ZHANG Yiyu;DING Wenfei;WANG Lei;TANG Xue;SONG Shuaifei;YAO Hui;HUANG Deli;XU Dejun;ZHAO Zhongquan(College of Animal Science and Technology,Southwest University,Chongqing 400715,China;Chongqing Dazu Agricultural Technical Service Center,Chongqing 402360,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2023年第5期1035-1043,共9页
Chinese Journal of Veterinary Science
基金
重庆市技术创新与应用发展专项基金资助项目(cstc2021jscx-gksbX0008)
