细胞因子信号传导抑制蛋白-1减轻细胞因子诱导胰岛β细胞促凋亡基因的表达  

SOCS-1 reduced the cytokine-induced pro-apoptosis gene expression in pancreatic β cells

作　　者：孙琦[1] 向若兰[2] 杨燕丽[3] 冯凯[1] 张文龙[1] 杨国华[1] 宋爱玲[1] 

机构地区：[1]中国医学科学院北京协和医学院北京协和医院内分泌科卫生部内分泌重点实验室,北京100730 [2]北京大学医学部生理与病理生理学系,北京100191 [3]中国医学科学院北京协和医学院北京协和医院内科,北京100730

出　　处：《基础医学与临床》2010年第12期1278-1283,共6页Basic and Clinical Medicine

基　　金：北京市自然科学基金(7082076)

摘　　要：目的构建稳定表达细胞因子信号传导抑制蛋白-1(SOCS-1)蛋白的细胞,探讨其在胰岛β细胞凋亡中的保护作用。方法克隆大鼠SOCS-1基因,构建表达载体pEGFP-C1-SOCS1并转染入大鼠胰岛细胞系RINm5F中,构建稳定表达SOCS-1蛋白的细胞。分别用细胞因子IL-1β、TNF-α、IFN-γ、IL-1β联合IFN-γ、IL-1β联合TNF-α及IFN-γ刺激24 h,RT-PCR与Western blot检测凋亡相关指标。结果成功构建了稳定表达SOCS-1蛋白的细胞。用IFN-γ、IL-1β联合IFN-γ、IL-1β联合TNF-α及IFN-γ刺激后,未转染SOCS-1质粒的细胞iNOS转录水平显著升高。在用所有细胞因子刺激后,未转染SOCS-1质粒的细胞caspase-3表达及活性均显著升高。结论 SOCS-1蛋白能减轻多种细胞因子诱导的胰岛β细胞促凋亡基因的表达。Objective To generate a cell line overexpressing SOCS1 protein and to investigate the protective effect of SOCS-1 on apoptosis of pancreatic β cell.Methods The rat SOCS-1 coding sequence was amplified and subcloned into pEGFP-C1 vector.The recombinant plasmid pEGFP-C1-SOCS1 was transfected into RINm5F cell line（insulin-secreting tumor cell line）.We generated a rat RINm5F cell line overexpressing recombinant plasmid pEGFP-C1-SOCS1.Cells were treated by several cytokines,including TNF-α,IL-1β,IFN-γ,IL-1β＋IFN-γ,IL-1β＋TNF-α＋IFN-γ.The cellular apoptosis were measured by RT-PCR and Western blot.Results We have generated a rat RINm5F cell line overexpressing SOCS1 protein successfully.The expression level of the iNOS mRNA was significantly increased in RINm5F cells after treatment by IFN-γ,IL-1β＋IFN-γ,IL-1β＋TNF-α＋IFN-γ,but notSOCS1-RINm5F cells.The expression and activity of caspase-3 was significantly increased in RINm5F cells after treatment by all cytokines.The caspase-3 expression and activity were not significantly changed in SOCS1-RINm5F cells after treated by cytokines.Conclusion Overexpression of SOCS-1 can reduce cytokine-induced pro-apoptosis gene expression in pancreatic β cells.

关 键 词：细胞因子信号传导抑制蛋白-1 胰岛细胞 细胞因子 凋亡 

分 类 号：R587.1[医药卫生—内分泌]