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14-3-3 epsilon 在睾丸生殖细胞凋亡中发挥重要作用:实验性精索静脉曲张的功能蛋白质组学研究。

14-3-3 epsilon plays an important role in testicular germ cell apoptosis: A functional proteomic study of experimental varicocele.

机构信息

Department of Infertility and Sexual Medicine, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, China.

Department of Urology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

出版信息

Andrologia. 2019 Jul;51(6):e13275. doi: 10.1111/and.13275. Epub 2019 Apr 5.

Abstract

The latest perspective indicates that apoptotic dysregulation is an important mechanism in male infertility induced by varicocele. To elucidate the molecular mechanism of apoptosis caused by varicocele, we used proteomics (2D-MALDI-TOF MS) to identify the altered proteins in the testes of experimental varicocele rats compared with the control. Here, 21 significantly different protein spots were detected by proteomics technology. 14-3-3 epsilon (14-3-3ε) was our subsequent research target because of its function in apoptosis. The expression of 14-3-3ε in rat testes was confirmed by Western blot and immunohistochemistry, and the terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) method was used to analyse the apoptosis of germ cells. GC-1 spg cells transfected with small interfering RNA were used to confirm the function of 14-3-3ε in vitro. 14-3-3ε protein expression decreased, accompanied by a higher apoptosis index in rat testes of the varicocele group. Furthermore, 14-3-3ε siRNA-treated GC-1 spg cells caused the upregulation of the apoptotic rate detected by flow cytometry. The expression of Bax and Bcl-2 was found to be regulated by 14-3-3ε in vitro. Our investigation demonstrated the pro-apoptotic function of the downregulation of 14-3-3ε, which may play an important role in germ cell apoptosis induced by varicocele.

摘要

最新观点表明,细胞凋亡失调是精索静脉曲张导致男性不育的一个重要机制。为了阐明精索静脉曲张导致细胞凋亡的分子机制,我们使用蛋白质组学(2D-MALDI-TOF MS)技术比较了实验性精索静脉曲张大鼠睾丸与对照组之间发生改变的蛋白质。通过蛋白质组学技术检测到 21 个明显不同的蛋白质斑点。14-3-3 ɛ(14-3-3ε)是我们后续研究的目标,因为它在细胞凋亡中发挥作用。通过 Western blot 和免疫组织化学法证实了大鼠睾丸中 14-3-3ε的表达,并使用末端脱氧核苷酸转移酶 dUTP 缺口末端标记法(TUNEL)分析生精细胞的凋亡。使用小干扰 RNA 转染 GC-1 spg 细胞,以确认 14-3-3ε在体外的功能。精索静脉曲张组大鼠睾丸中 14-3-3ε蛋白表达降低,同时凋亡指数升高。此外,流式细胞术检测到经 14-3-3ε siRNA 处理的 GC-1 spg 细胞凋亡率上调。体外研究发现 Bax 和 Bcl-2 的表达受 14-3-3ε调控。我们的研究表明下调 14-3-3ε具有促凋亡作用,这可能在精索静脉曲张导致的生精细胞凋亡中发挥重要作用。

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