CAJ | 학술논문

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。
목적：통과관찰고온대체외대서고환지지세포조망적영향，탐토고온재남성생식공능강저혹불육중적작용궤제。방법분리배양대서고환지지세포（Sertoli cell，SC）병감정。실험분위실험조（39℃하배양2h、4h、8h、12h급24h）화대조조（36℃）。CCK-8법검측 SC증식활성，류식세포의검측SC조망솔，RT-PCR검측Fas/Fasl급Bax/Bcl-2기인전록수평。결과본실험중， SC순도체도96.15%；수고온시간적연장，SC증식활성축점강저（P ＜0.05）；조망솔정현선승고후강저적추세，재고온작용하8h조망솔최고（P ＜0.05）； Fas、Fasl、Bax기인전록수평축점승고（P＜0.05），Bcl-2기인전록수평축점하강（P＜0.01），차정시간효응의뢰관계。결론고온도치SC증식능력강저，병가능통과“사망수체”화“선립체”량조신호전도도경도치SC조망，파배생정미배경，진이도치남성생식공능강저심지불육。
Objective To explore the pathological mechanism of heat stress in male reproduction ability reducing or male infertility. Methods Sertoli cells were isolated from the testicle of male Wistar 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.