摘要
目的通过观察高温对体外大鼠睾丸支持细胞凋亡的影响，探讨高温在男性生殖功能降低或不育中的作用机制。方法分离培养大鼠睾丸支持细胞（Sertoli cell，SC）并鉴定。实验分为实验组（39℃下培养2h、4h、8h、12h及24h）和对照组（36℃）。CCK-8法检测SC增殖活性，流式细胞仪检测SC凋亡率，RT-PCR检测Fas／Fasl及Bax／Bcl-2基因转录水平。结果本实验中，SC纯度达到96．15％；随高温时间的延长，SC增殖活性逐渐降低（P〈0．05）；凋亡率呈现先升高后降低的趋势，在高温作用下8h凋亡率最高（P〈0．05）；Fas、Fasl、Bax基因转录水平逐渐升高（P〈0．05），Bcl-2基因转录水平逐渐下降（P〈0．01），且呈时间效应依赖关系。结论高温导致SC增殖能力降低，并可能通过“死亡受体”和“线粒体”两条信号传导途径导致SC凋亡，破坏生精微环境，进而导致男性生殖功能降低甚至不育。
Objective To explore the pathological mechanism reducing or male infertility. Methods Sertoli cells were isolated from of heat stress in male reproduction ability the testicle of male VCistar rat and identified by Fasl. The SCs were divided into the experimental group （39℃ 2, 4, 8, 12, 24h） and the control group （36℃） Proliferation activity of SCs was determinated by CCK-8 assay, then the apoptosis ratio of SCs was determined with flow cytometry （FCM）, and the level of Fas/Fasl and Bax/Bcl-2 were assessed by RT-PCR. Results The purity of SCs was 96.15%. The proliferation of SCs was lower than that of the control under exposure to high temperature of 39℃ at any time length（P〈0.05）; The apoptotic ratios of SCs were higher in exposure to heat stress for 2 to 8 hours than that of the control（P〈0.05）, but declined after longer exposure to under heat stress （P〈0.01）; Compared with that of the control group, the expressions of Fas, Fasl and Bax in the experimental group increased along with exposure time grow （P〈0.05）, but the Bcl-2 declined （P〈0.01）. Conclusion Heat stress results in the decreas of proliferative ability of SCs in vitro, and increases the apoptosis of SCs through the two singnal transduction pathways： death receptor pathway and mitochondrial pathway. It suggests that high temperature might impare the male spermatogensis or male fertility by inducing the apoptosis of SCs.
出处
《中国男科学杂志》
CAS
CSCD
北大核心
2013年第8期16-20,共5页
Chinese Journal of Andrology
关键词
高温
支持细胞
细胞凋亡
heat stress
sertoli cells apoptosis
