摘要
目的探讨敲低热休克转录因子5(Hsf5)对小鼠睾丸间质细胞(TM3)和支持细胞(TM4)中热休克家族表达的影响。方法培养小鼠TM3和TM4细胞,随机分为对照组(control group)和Hsf5敲低组(Hsf5 knockdown group)。用实时荧光定量PCR(RT-qPCR)检测两种细胞中热休克转录因子(HSFs)和热休克蛋白(HSPs)mRNA的表达。根据RT-qPCR结果分别选取两种细胞中变化具有显著差异的HSPs(P<0.01),应用Western blot检测TM3细胞中HSPA2、HSPA5、HSP90ab1和TM4细胞中HSPA2、HSP90aa1蛋白表达。结果用siRNA干扰技术成功构建Hsf5敲低模型。敲低Hsf5后,TM3细胞中Hspa2、Hspa5、Hsp90ab1和TM4细胞中Hsf2、Hspa2、Hsp90aa1、Hsp90ab1、Hspd1的mRNA表达明显下调(P<0.05)。与对照组相比,在Hsf5敲低组中,TM3细胞中HSPA2、HSPA5和TM4细胞中HSPA2蛋白表达明显下调(P<0.05)。结论HSF5可能通过调控小鼠睾丸间质细胞和支持细胞中HSPA2和HSPA5的表达参与精子发生。
Objective To investigate the effect of knockdown heat shock transcription factor 5(Hsf5)on the expression of heat shock family in mouse Leydig cells(TM3)and Sertoli cells(TM4).Methods Mouse TM3 and TM4 cells were cultured and randomly divided into control group and Hsf5 knockdown group.The expression of heat shock transcription factor(HSF)and heat shock protein(HSP)mRNA in the two groups of cells was detected by real-time quantitative PCR.According to the results of RT-qPCR,HSPs with significant differences in the two kinds of cells were selected(P<0.01).Western blot were used to detect the expression of HSPA2,HSPA5,HSP90ab1 in TM3 cells and HSPA2,HSP90aa1 in TM4 cells.Results Hsf5 knockdown model was successfully constructed by RNA interference.After knockdown of Hsf5,the mRNA expression of Hspa2,Hspa5,Hsp90ab1 in TM3 cells and Hsf2,Hspa2,Hsp90aa1,Hsp90ab1,Hspd1 in TM4 cells was all significantly down-regulated(P<0.05).In Hsf5 knockdown group,the expression of HSPA2,HSPA5 proteins in TM3 cells and HSPA2 protein in TM4 cells were significantly down-regulated(P<0.05).Conclusions HSF5 is potentially involved inspermatogenesis by regulating the expression of HSPA2 and HSPA5 in mouse Leydig cells and Sertoli cells.
作者
钟琳
钟洪莉
代宇婕
张庆华
ZHONG Lin;ZHONG Hongli;DAI Yujie;ZHANG Qinghua(Department of Obstetrics and Gynecology,Army Medical Center of PLA,Army Medical University,Chongqing 400042,China)
出处
《基础医学与临床》
2023年第4期626-631,共6页
Basic and Clinical Medicine
基金
2021年度大学指令性计生专项科研课题(2021XZL06)。
关键词
热休克转录因子5
热休克转录因子
热休克蛋白
精子发生
heat shock transcription factor 5
heat shock transcription factors
heat shock proteins
spermatogenesis
