干扰EZH2的表达对奶山羊精原干细胞抗氧化能力的影响  

Effect of EZH2 knockdown on antioxidant capacity of goat male germline stem cells

作　　者：蔡玉 邓明田 刘孜斐 张国敏[1] 马键宇 安世钰 王智博[1] 张艳丽[1] 王锋[1] CAI Yu;DENG Mingtian;LIU Zifei;ZHANG Guomin;MA Jianyu;AN Shiyu;WANG Zhibo;ZHANG Yanli;WANG Feng(Jiangsu Livestock Engineering Laboratory,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学江苏省家畜胚胎工程实验室,江苏南京210095

出　　处：《南京农业大学学报》2021年第4期726-732,共7页Journal of Nanjing Agricultural University

基　　金：国家重点研发计划项目(2018YFD0501900)。

摘　　要：[目的]本试验旨在探究EZH2对奶山羊精原干细胞抗氧化能力的影响。[方法]通过siRNA干扰EZH2在奶山羊精原干细胞中的表达,验证干扰效率,并采用ELISA、qPCR和Western blot检测抗氧化酶活性和抗氧化相关基因与蛋白表达水平。[结果]与对照组相比,干扰EZH2极显著提高奶山羊精原干细胞中过氧化氢酶(CAT)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GPx)的活性(P<0.01),而丙二醛(MDA)含量极显著下降(P<0.01)。在分子水平上,干扰EZH2后,奶山羊精原干细胞中CAT、SOD2蛋白和mRNA表达水平均极显著提高(P<0.01),GPx mRNA表达水平极显著上升(P<0.01)。[结论]在体外培养的奶山羊精原干细胞中,干扰EZH2增强了细胞的抗氧化能力,其分子机制可能与激活CAT和SOD2蛋白并促进其下游抗氧化基因GPx表达相关。[Objectives]The experiment was conducted to investigate the role of enhancer of zeste homolog 2(EZH2)on the antioxidant capacity of goat male germline stem cells(mGSC).[Methods]Small interference RNA(siRNA)was used to knockdown the expression of EZH2 in mGSC.After verifying the effective siRNA,the enzymic activity of catalase(CAT),superoxide dismutase(SOD),glutathione peroxidase(GPx)and the content of malondialdehyde(MDA)were detected by ELISA.Then,the expression levels of antioxidant indicators including CAT,SOD2 and GPx in goat mGSC were detected by qPCR and Western blot.[Results]The ELISA results showed that the enzymic activities of CAT,SOD and GPx significantly increased(P<0.01),but the content of MDA significantly decreased(P<0.01)in mGSCs after knockdown of EZH2 compared with the control group.The mRNA and protein expression levels of CAT and SOD2 significantly increased(P<0.01),and the transcriptional level of GPx significantly increased(P<0.01)in EZH2 knockdown mGSC compared with the control group.[Conclusions]EZH2 knockdown increased the antioxidant capacity of mGSC,and its molecular mechanism might be related to the activation of oxidative stress-related genes.

关 键 词：山羊 睾丸 精原干细胞 EZH2 抗氧化能力 

分 类 号：S826.8[农业科学—畜牧学]