己烯雌酚染毒对小鼠睾丸引带细胞纤维肌动蛋白和游离钙离子浓度的影响  

Effects of diethylstilbestrol on F-actin and free Ca^(2+) in mouse gubernaculum cells in culture

作　　者：张镟[1] 蒋学武[1] 李建宏[1] 段守兴[1] 马廉[2] 

机构地区：[1]汕头大学医学院第二附属医院小儿外科,广东汕头515041 [2]汕头大学医学院第二附属医院儿科

出　　处：《环境与健康杂志》2014年第2期100-103,共4页Journal of Environment and Health

基　　金：国家自然科学基金(81341099;81170555)

摘　　要：目的通过检测己烯雌酚(diethylstilbestrol,DES)对小鼠睾丸引带细胞纤维肌动蛋白(F-actin)和游离钙离子浓度([Ca2+]i)的影响,从细胞水平探讨外源性雌激素对小鼠睾丸引带细胞收缩功能的影响及其可能机制。方法将小鼠睾丸引带细胞分别培养于终浓度为0(溶剂对照)、0.01、0.10、1.00、10.00μg/ml的含DES无血清培养基中24 h。采用细胞免疫荧光法检测细胞F-actin的表达情况,采用共聚焦激光扫描显微镜检测细胞[Ca2+]i的变化。结果经DES处理后的小鼠睾丸引带细胞质周边的肌动蛋白丝模糊,细胞内出现明显粗壮的束状纤维,结构紊乱。与溶剂对照组比较,各浓度DES染毒小鼠睾丸引带细胞内[Ca2+]i均较高,差异有统计学意义(P<0.05);且随着DES染毒浓度的升高,小鼠睾丸引带细胞内[Ca2+]i呈上升趋势。结论除传统的雌激素核受体介导的信号传导途径外,DES还可能通过非基因调控信号途径快速增加细胞内钙离子浓度,使由F-actin组成的纤维排列紊乱,并可能导致睾丸引带细胞结构紊乱,收缩功能减弱。Objective To observe the F-actin and free Ca^2＋（[Ca^2＋]i） in Kunming mice guberuacular cells in culture, and to explore the effects of exogenous estrogens （EEs） on testis gubernaculum and testis development by investigating the effects of gubernaeular cells exposed to diethylstilbestrol on the expressions of contractility. Methods Mouse gubernaculums testis subcuhuring cells were divided into six groups randomly, such as, normal group （not given any drug） , control group （given DMSO, 1 μl/ml）, and experimental groups 1-4：given 0.01, 0.10, 1.00 and 10.00 μg/ml diethylstilbestrol respectively （diethylstilbestrol dissolved in DMSO）. The expression of F-actin was observed by immunofluorescence. Fluo-3/AM and Pluronic F-127 were loaded as Ca2＋ sensitive probe. The changes of intracellular [Ca^2＋]i were observed by confocal laser scanning microscopy. Results Diethylstilbestrol exhibited the stress fibres distributed throughout the cytoplasm gradually instead of the F-actin around the cells, especially in the high dose. Diethylstilbestrol increased fluo-3/AM fluorescence in guberuaculums testis cells in dose dependent manner, with statistically significant differences （P〈0.05）. Conclusion Diethylstilbestrol exposure may increase intracellular [Ca^2＋]i via non-genomic signaling and induce F-actin rearrangement and stress fiber formation in gubernaculum testis cells. Diethylstilbestrol may inhibit the cell morphosis and contractility of gubernaculum.

关 键 词：己烯雌酚 外源性雌激素 睾丸引带 游离钙离子浓度 纤维肌动蛋白 

分 类 号：R994.6[医药卫生—毒理学]