热休克蛋白70-2基因沉默对大鼠精子附睾成熟的影响

目的 观察热休克蛋白(HSP)70-2基因沉默对大鼠附睾中精子成熟的影响.方法 构建HSP70-2特异的短发夹RNA(shRNA)质粒载体,注入大鼠附睾中,1周后处死大鼠,无菌切取附睾,用逆转录-聚合酶链反应(RT-PCR)方法 检测附睾中HSP70-2 mRNA表达,Western blot观察HSP70-2蛋白表达,应用苏木素-伊红(HE)染色检测附睾管内精子密度.结果 空白对照组HSP70-2蛋白表达量为1.31±0.10,附睾管内成熟精子密度为(63.46±14.43)%.基因沉默组HSP70-2蛋白表达量为0.93±0.13,附睾管内成熟精子密度为(31.51±10.67)%,与对照组比较均显著减少,差异有统计学意义(P＜0.01).结论 HSP70-2基因沉默影响附睾中精子成熟.

Effects of HSP70-2 gene silencing on the maturation of rat sperms in epididymis

Objective To investigate the effects of heat shock protein (HSP) 70-2 small interfer-ence RNA (siRNA) on the maturation of rat sperms in epididymis. Methods The recombinant plasmid series of HSPT0-2-targeted short hairpin RNA (shRNA) were constructed by pGenesil-1 plasmids vector, and injected into rat epididymis. The HSP70-2 expression was detected by RT-PCR and Western blot. The sperm density was evaluated by HE staining. Results The HSP70-2 protein expression was 1.31 ± 0.10, and the mature sperm density in epididymis was (63.46 ± 14.43) % in the control group. Those were 0.93 ± 0.13 and (31.51 ± 10.67)% in the pGenesil-HSP70-2 treated group, respectively. There was sig-nificant difference between the two groups (P ＜ 0.01). Conclusion HSP70-2-shRNA could significantly affect the maturation of rat sperms in epididymis.