血管平滑肌细胞中睾酮膜受体的研究  

Membrane testosterone receptors in cultured vascular smooth muscle cells

作　　者：马瑞[1] 江时森[1] 程训民[1] 宫剑滨[1] 张启高[1] 林其谁[2] 

机构地区：[1]南京军区南京总医院心血管内科,江苏南京210002 [2]中国科学院上海生物化学与细胞生物学研究所,上海200031

出　　处：《中华男科学杂志》2010年第5期428-432,共5页National Journal of Andrology

摘　　要：目的:观察血管平滑肌细胞(VSMCs)胞膜有无睾酮(T)特异性结合位点(膜受体),并对其与经典的细胞内雄激素受体(iAR)的关系作初步探讨。方法:贴块法培养雄性SD大鼠胸主动脉VSMCs,给以异硫氰酸荧光素(FITC)标记的牛血清白蛋白-睾酮复合物(T-BSA-FITC)作用,流式细胞仪检测VSMCs荧光标记情况,以及iAR拮抗剂氟他胺对T-BSA-FITC与VSMCs结合的抑制作用;用免疫荧光法标记胞膜完整和TritonX-100处理后胞膜透化VSMCs表达的iAR,流式细胞术检测细胞平均荧光强度,进一步转换为相对荧光强度进行统计学分析。结果:与0作用时间点相比,T-BSA-FITC与VSMCs作用10s,细胞相对荧光强度即显著增加(P<0.01);VSMCs与T-BSA-FITC作用10min,细胞相对荧光强度显著高于对应摩尔浓度的BSA-FITC、BSA-FITC+BSA、BSA-FITC+BSA+T对照(P值分别为0.357,0.832和0.823)。与单纯T-BSA-FITC作用相比,预先给以iAR拮抗剂氟他胺对VSMCs荧光强度没有显著影响(P=0.318)。胞膜完整VSMCs与iAR抗体作用后,细胞相对荧光强度较单纯加入FITC标记二抗无明显增加(P=0.733);而胞膜透化VSMCs与iAR抗体作用后,细胞荧光强度较单纯FITC标记二抗处理明显增加(P=0.024)。结论:VSMCs中存在睾酮膜受体,其性质不同于经典的iAR。Objective：To determine the presence of membrane testosterone receptors in cultured vascular smooth muscle cells （VSMC）,and invertigate their relationship with classical intracellular androgen receptors （iAR）.Methods：VSMCs were cultured from the thoracic aorta of male Sprague-Dawley rats by the explant method.Subconfluent VSMCs were incubated with serum-free medium for 24 h to obtain quiescent non-dividing cells,and then treated with the indicated agents.The aliquots of VSMCs were labeled with testosterone-BSA-FITC （T-BSA-FITC） and analyzed by flow cytometry.Classical iARs in intact-and permeabilized-cells were detected with anti-iAR antibodies and FITC-labeled secondary antibodies by immunofluorescence,followed by flow cytometry analysis.Results：Incubation of VSMCs with T-BSA-FITC obviously increased their relative fluorescence intensity at 10 sec as compared with the untreated controls （P0.01）,and so did it at 10 min in comparison with the treatment with BSA-FITC alone or together with free testosterone （P0.01）.Pretreatment with iAR antagonist flutamide exhibited no significant influence on the relative fluorescence intensity of VSMCs （P = 0.318）.Traditional iARs were not detectable on the surface of intact VSMCs,although permeabilized cells contained iARs.Conclusion：VSMCs contain testosterone receptors in the plasma membrane,and these membrane receptors are not identical to classical iARs.

关 键 词：血管平滑肌 睾酮 雄激素受体 膜受体 

分 类 号：R541.4[医药卫生—心血管疾病]