MGRN1对小鼠睾丸精原干细胞线粒体自噬的影响  

MGRN1 affects the mitophagy of spermatogonial stem cells in mice

作　　者：朱元[1] 罗勇[2] 刘发英[2] 汪利群[1] ZHU Yuan;LUO Yong;LIU Fa-ying;WANG Li-qun(Department of Reproductive Health,Jiangxi Hospital for Women and Children,Nanchang,Jiangxi 330006,China;Central Laboratory,Jiangxi Hospital for Women and Children,Nanchang,Jiangxi 330006,China)

机构地区：[1]江西省妇幼保健院生殖健康科,江西南昌330006 [2]江西省妇幼保健院中心实验室,江西南昌330006

出　　处：《中华男科学杂志》2021年第11期969-973,共5页National Journal of Andrology

基　　金：国家自然科学基金(81660265);江西省重点研发计划(20202BBGL73065);江西省自然科学基金(20181BAB205015);江西省卫生计生委科技计划(20175430,20191104)。

摘　　要：目的:研究MGRN1对小鼠睾丸精原干细胞(SSCs)线粒体自噬的影响。方法:体外培养小鼠睾丸精原干细胞,分为3组,对照组(空载体组)、MGRN1组(利用RNAi下调SSCs中MGRN1的表达)、MGRN1+FCCP组(在MGRN1下调后的SSCs中加FCCP诱导线粒体自噬);利用Western印迹检测3组线粒体功能相关蛋白(Cytochromo c、COX IV)及线粒体自噬相关蛋白LC3、P62、CK2表达及FUNDC1磷酸化水平。电镜观察各组SSCs的线粒体及线粒体自噬小体。结果:免疫印迹显示,MGRN1组及MGRN1+FCCP组与对照组相比,Cytochromo c、COX IV、LC3、P62表达明显下降,FUNDC1磷酸化水平增加,促FUNDC1磷酸化的蛋白CK2表达增加(P<0.05)。而MGRN1+FCCP组和MGRN1组相比,MGRN1、Cytochrome c、COX IV、LC3、P62、CK2表达及FUNDC1磷酸化无明显差异(P>0.05)。电镜显示:与对照组相比MGRN1组及MGRN1+FCCP组中,SSCs的线粒体损伤增加,线粒体自噬小体减少。结论:MGRN1下调后,体外培养的小鼠SSCs中线粒体受损、线粒体自噬受抑制、加FCCP也无法诱导出线粒体自噬增加,即MGRN1影响小鼠睾丸线粒体自噬,可能与CK2影响FUNDC1的磷酸化有关,具体分子机制有待进一步研究。Objective:To study the effect of mahogunin ring finger-1(MGRN1)on the mitophagy of the spermatogonial stem cells(SSC)in mice.Methods:SSCs cultured in vitro were divided into three groups:empty vector control,MGRN1(MGRN1 in SSCs knocked down by RNAi),and MGRN1+FCCP(inducing mitophagy with carbonyl cyanide p-trifluoromethoxyphenylhydrazone[FCCP]in the SSCs with down-regulated MGRN1).The expressions of mitochondrial function-related proteins(Cytochromo c and COX IV)and mitophagy-related proteins(LC3,P62,FUNDC1 and CK2)and the phosphorylation of FUNDC1 were detected by Western blot.Mitochondria and mitochondrial autophagosomes in the SSCs were observed under the electron microscope.Results:Compared with the empty vector control group,the MGRN1 and MGRN1+FCCP groups showed significantly down-regulated expressions of Cytochromo c,Cox IV,LC3 and P62,increased phosphorylation level of FUNDC1,and up-regulated expression of CK2 in the SSCs(P<0.05).No statistically significant differences were found in the expressions of Cytochromo c,Cox IV,LC3,P62 and CK2 or in the phosphorylation level of FUNDC1 between the MGRN1 and MGRN1+FCCP groups(P>0.05).Electron microscopy manifested increased mitochondrial damage and reduced mitochondrial autophagosomes in the SSCs in the MGRN1 and MGRN1+FCCP groups compared with those in the control group.Conclusion:MGRN1 affects mitophagy in the SSCs of mice,which may be associated with the effect of CK2 on the phosphorylation of FUNDC1,and its molecular mechanism needs to be further studied.

关 键 词：MGRN1 小鼠睾丸精原干细胞 线粒体自噬 

分 类 号：R698.2[医药卫生—泌尿科学]