小鼠睾丸支持细胞Attractin基因的抗氧化和抗凋亡作用的初步研究  

Antioxidative and antiapoptotic effects of the Attractin gene on Sertoli cells in mice

作　　者：李洁[1] 王清 程丹[1] 杨菁[1] 熊承良[3] 

机构地区：[1]武汉大学人民医院生殖医学中心 [2]解放军95171部队计划生育办公室 [3]华中科技大学同济医学院计划生育研究所

出　　处：《中华男科学杂志》2014年第9期787-791,共5页National Journal of Andrology

基　　金：国家自然科学基金(30570679;81401225)~~

摘　　要：目的:观察Attractin(Atrn)基因不同程度缺失对小鼠睾丸支持细胞抗氧化和抗凋亡的作用,及其对细胞超氧化物歧化酶(SOD)、caspase 6表达的影响。方法:观察Atrn基因正常表达、部分缺失、完全缺失的三组细胞(psiRNA-TM4、psiAtrn-TM4、mu-SC)的细胞凋亡指数;采用荧光定量PCR、Western印迹技术检测SOD、caspase 6mRNA及蛋白表达水平;采用化学比色法检测细胞中SOD活性和丙二醛(MDA)含量。结果:与对照组psiRNATM4比较,抑制Atrn基因的表达后,支持细胞的SOD mRNA表达水平下降,psiAtrn-TM4、mu-Sc组细胞SOD mRNA表达量分别下降了70.76%和92.58%,caspase 6 mRNA表达水平上升,分别为对照组的5.28倍和2.97倍;蛋白水平的变化趋势与mRNA结果一致,psiAtrn-TM4及mu-Sc组细胞中SOD蛋白的表达分别下降了65.11%和71.0%,caspase 6蛋白表达分别为对照组的3.4倍和2.5倍,差异均有统计学意义(P<0.05)。抑制Atrn基因表达后,psiAtrn-TM4、mu-SC组细胞凋亡率分别增加16.22%和22.03%,SOD活性分别下降23.00%和39.37%,mu-SC组MDA含量增加155.22%,差异有统计学意义(P<0.05)。结论:Atrn基因对睾丸支持细胞功能的影响是通过多途径实现的,抗氧化应激以及调控凋亡途径可能在其中起着重要作用。Objective： To evaluate the effects of Attractin（Atrn） silence on the anti-oxidative and anti-apoptotic abilities of TM4 Sertoli cells and its influence on the expressions of superoxide dismutase （SOD） and caspase6 in the cells. Methods： We observed the apoptotic indexes of TM4 Sertoli cells with normal expression （ control）, partial deletion, and complete deletion of the Atrn gene （psiRNA-TM4, psiAtm-TM4, and mu-SC）. We determined the mRNA and protein expressions of SOD and caspase6 by Q-PCR and Western blot, measured the SOD activity andmalondialdehyde（MDA） contentby spectrophotometry, and detected the apoptotic index of the cells by TUNEL. Results ： Compared with psiRNA-TM4, after inhibition of the Atrn expression, the Sertoli cells in the psiAtrn- TM4 and mu-SCgroupsshowed significantly decreased expressions ofSOD mRNA（70.76% and 92.58% ） and protein （65.11% and 71.0% ） （bothP 〈 0.05 ）. The levels ofcaspase 6 mRNA and protein were increased 5.28 and 3.40 times in the psiAtrn-TM4and 2.97 and 2.50 times in the mu-SCgroup as compared with the normal control （ both P 〈 O. 05）. Atrn deletion markedly increased the apop-totic indexesof thecells in the psiAtrn-TM4 and mu-SCgroupsby 16.22% and 22.03 % （P 〈 0.05） and reduced the activity of SOD by 23.00% and 39.37% （P 〈 0.05） ; it also elevated the level of MDAby 155.22% （P 〈 0.05）. Conclusion ： The Atrn gene exerts influence on the function of Sertoli cells inmuhiple ways, in whichantioxidative stress and apoptosis regulation may play an important role.

关 键 词：ATTRACTIN 凋亡 CASPASE 6 支持细胞 小鼠 超氧化物歧化酶 

分 类 号：R346[医药卫生—基础医学]