基础医学与临床 ›› 2010, Vol. 30 ›› Issue (12): 1278-1283.

• 研究论文 • 上一篇    下一篇

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

孙琦1,向若兰2,杨燕丽3,冯凯4,张文龙1,杨国华1,宋爱玲1   

  1. 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2. 北京大学医学部生理学与病理生理学系3. 中国医学科学院北京协和医学院北京协和医院4. 中国医学科学院 北京协和医学院 北京协和医院
  • 收稿日期:2010-10-12 修回日期:2010-10-18 出版日期:2010-12-05 发布日期:2010-12-05
  • 通讯作者: 孙琦

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

SUN Qi 1,XIANG Ruo-lan 2,YANG Yan-li 3,FENG Kai 1,ZHANG Wen-long 1,YANG Guo-hua 1,SONG Ai-ling 1   

  1. 1. PUMC Hospital, CAMS & PUMC2. Department of Physiology and Pathophysiology, Peking University Health Science Center3. Peking Union Medical College Hospital, CAMS & PUMC
  • Received:2010-10-12 Revised:2010-10-18 Online:2010-12-05 Published:2010-12-05
  • Contact: SUN Qi

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-r、IL-1β联合TNF-α及IFN-γ刺激24h,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-r、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。

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

Abstract: Objective This paper aims to generate a cell line overexpressing SOCS1 protein and 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-r, IL-1β+IFN-r, IL-1β+TNF-α+IFN-r. 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 treated by IFN-r, IL-1β+IFN-r, IL-1β+TNF-α+IFN-r, but not SOCS1-RINm5F cells. Western blot analysis demonstrated that the expression and activity of caspase-3 was significantly increased in RINm5F cells after treated by all cytokines. However, the caspase-3 expression and activity was not significantly different in SOCS1-RINm5F cells after treated by cytokines. Conlusion Overexpression of SOCS-1 can reduced cytokine-induced pro-apoptotic gene expression in pancreatic β cells.

中图分类号: