α-硫辛酸对氧化应激下睾丸支持细胞骨架蛋白表达的影响  

Effects of α-1 ipoic acid on the expressions of cytoskeleton proteins in the oxidative stress-induced sertoli cells

作　　者：张荣秋[1] 王英俊[1] 徐敏[1] 王高能[1] 张唯力[1] 

机构地区：[1]重庆医科大学附属第二医院泌尿外科,重庆400010

出　　处：《中国男科学杂志》2011年第10期17-22,共6页Chinese Journal of Andrology

摘　　要：目的探讨α-硫辛酸（α-lipoicacid，α—LA）对氧化应激下大鼠睾丸支持细胞骨架蛋白表达的影响。方法原代培养大鼠睾丸支持细胞，分为4组：对照组、LA组、H202组及H202＋LA组。CCK．8法检测各组细胞存活率；DCFH—DA荧光探针法检测细胞内活性氧水平：酶活性检测试剂盒检测抗氧化酶活性；免疫组织化学法、蛋白质免疫印迹法检测波形蛋白及微管蛋白的表达。结果与对照组比较，H2O2刺激后支持细胞存活率下降（P〈0．05），细胞内ROS水平上升（P〈0．05），抗氧化酶活性降低（P〈0．05）；波形蛋白及微管蛋白的表达也显著降低（P〈0．05），而LA干预后可提高细胞存活率，降低细胞内ROS水平，提高抗氧化酶的活性，并提高波形蛋白及微管蛋白的表达。结论氧化应激能降低支持细胞波形蛋白及微管蛋白表达，使细胞存活率下降；LA通过清除细胞内ROS，提高抗氧化酶的活性及波形蛋白和微管蛋白表达而对氧化应激的支持细胞起保护作用。Objective To explore the effects of α -lipoic acid on the expression of cytoskeleton proteins in the oxidative stress-induced sertoli cells. Methods Primary rat sertoli cells were randomly divided into four groups including control group, LA（200μmol/L） group, H2O2（400 μmol/L） groupand H2O2 plus LA pretreatment group. Cell viability was assessed by CCK-8. Intracellular reactive oxygen species （ROS） were measured by the fluorescent probe DCFH-DA. The activities of antioxidant enzymes were measured by enzyme activity detection kit. The expressions of vimentin and α-tubulin were detected by immunohistochemistry and Western blot. Results Compared with those of the control group, cell viability was decreased, intracellular ROS was increased, activities of antioxidant enzymes were decreased and the expressions of vimentin and a-tubulin were downregulated in sertoli cells after the stimulation of H2O2（P〈0.05）. However, by LA pretreatment, cell viability was improved, activities of antioxidant enzymes were enhanced, intracellular ROS level was decreased, and the expressions of vimentin and a-tubulin.were upregulated （P 〈0.05 ）. Conclusion H2O2 might induce the expressions ofvimentin and a-tubulin in sertoli cells and its cell survival rate. The protective role of LA might amend this damage by eliminating intracellular ROS, improving the activities of antioxidant enzymes, and upregulating the expressions of vimentin and α-tubulin

关 键 词：睾丸 细胞骨架 氧化性应激 硫辛酸 

分 类 号：R697.22[医药卫生—泌尿科学]