GPR30在全氟辛烷磺酰基化合物诱导GC-1细胞凋亡中的作用  

The effects of GPR30 on apoptosis of GC-1 cells induced by PFOS

作　　者：瞿建华[1] 贲帅 仇梁林[1] 江俊康[1] 陈刚[1] 

机构地区：[1]南通大学公共卫生学院,江苏南通226019

出　　处：《毒理学杂志》2016年第5期349-352,353,共5页Journal of Toxicology

基　　金：国家自然科学基金(81302453;21277078;81302452);江苏省高等学校大学生创新创业训练计划项目(201510304066Y)

摘　　要：目的研究全氟辛烷磺酰基化合物(perfluorooctane sulfonate,PFOS)对生精细胞凋亡的影响,并探讨其可能机制及G蛋白偶联受体30(G protein-coupled receptor 30,GPR30)在此过程中的作用。方法以GC-1小鼠精原细胞株为染毒模型,设置对照组和PFOS染毒组(50、100和200μmol/L)。运用MTT法检测GC-1细胞的存活率;Annexin VFITC/PI双染色法检测GC-1细胞凋亡;采用蛋白质印迹技术(Western blot)检测不同剂量PFOS及同时加入GPR30特异性阻断剂G15后对生精细胞凋亡相关蛋白表达的影响。结果 PFOS能够显著抑制GC-1细胞的增殖、促进GC-1细胞凋亡;PFOS染毒后,GC-1细胞中GPR30及凋亡相关蛋白Cleaved caspase-3、Bax表达显著升高;G15阻断GPR30的表达后,PFOS对Cleaved caspase-3表达水平的提升作用被部分逆转。结论 PFOS能显著促进生精细胞凋亡,其机制可能是通过上调GPR30的表达继而激活线粒体凋亡通路,并最终影响精子发生。Objective To investigate the effects of perfluorooctane sulfonate（ PFOS） on apoptosis of germ cells,and discuss the possible mechanism and the role of G protein-coupled receptor 30（ GPR30） in the process. Methods GC-1 mouse spermatogonial cell line was treated with various concentrations of PFOS（ 0,50,100,200 μmol / L） for 24 h. MTT assay was used to assess cell viability and Annexin V-FITC / PI was performed to detect apoptosis of GC-1 cells. Western blot analysis was used to determine the expression of apoptosisrelated proteins of germ cells,which were treated with PFOS in the absent or in the present of GPR30 specific inhibitor G15. Results PFOS significantly inhibited the proliferation of GC-1 cells and promoted apoptosis of GC-1 cells. The expression levels of GPR30,cleaved caspase-3 and Bax in GC-1 cells significantly increased after the treatment of PFOS. The promoting effect of PFOS on Cleaved Caspase-3 protein expression was partially reversed when the expression of GPR30 was blocked by G15. Conclusion PFOS could significantly stimulate the apoptosis of GC-1 cells. This action might be mediated by GPR30,followed by activation of apoptotic pathway involving bax and cleaved caspase-3 up-regulation,which provides new information about the mechanism of PFOS-induced spermatogenic impairment.

关 键 词：PFOS 精子发生 GPR30 

分 类 号：R994.6[医药卫生—毒理学]