T细胞免疫球蛋白及黏蛋白家族-3对干扰素-γ活化的小鼠肝Kupffer细胞分泌炎性细胞因子的调节作用及机制  

Role of T cells immunoglobulin and mucin family-3 in regulation of cytokines secretion of mice liver Kupffer cell activated by interferon-γ

作　　者：牛坚[1] 王月[1] 李克清[1] 朱志军[2] 刘斌[1] 

机构地区：[1]徐州医学院附属医院,徐州市221004 [2]北京友谊医院普外科,北京100069

出　　处：《中华实验和临床感染病杂志（电子版）》2015年第5期130-133,共4页Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition)

基　　金：天晴甘美基金项目资助(No.CFHPC20132020);徐州市重大科研项目(No.KC14SX011)

摘　　要：目的探讨T细胞免疫球蛋白及黏蛋白家族-3(Tim3)对IFN-γ活化的小鼠Kupffer细胞的调节作用并探讨其相关机制。方法将真核表达质粒pc DNA3.1-Tim3转染小鼠肝Kupffer细胞,以Real-time PCR和Western blot检测Tim3在小鼠肝Kupffer细胞的表达。通过ELISA检测质粒pc DNA3.1-Tim3、Tim3阻断型抗体对IFN-γ活化的小鼠肝Kupffer细胞因子(TNF-α、IL-1β和IL-6)产生的影响,Western blot检测JAK2/STATl蛋白表达。结果 Real-time PCR检测结果显示,IFN-γ能够显著提高小鼠肝脏Kupffer细胞中Tim3 m RNA的表达水平(P<0.05);pc DNA3.1-Tim3组的TNF-α、IL-6和IL-1β分泌较对照组显著下降(P<0.01);ELISA结果显示,Tim3阻断型抗体组与对照组相比,TNF-α、IL-6和IL-1β的分泌增加(P<0.01);Western blot检测显示,与对照组相比,Tim3阻断型抗体预处理的小鼠Kupffer细胞JAK2及STAT1蛋白的表达上调(P<0.05)。结论Tim3通过调控Jak2/Stat1蛋白表达参与了Kupffer细胞活化的调节。Objective To investigate the adjustment role of T cells immunoglobulin and mucin family-3 （Tim 3） on Kupffer cells activation and the related mechanism. Methods The Tim3 pcDNA3.1-Tim3 plasmids were transfected into Kupffer cells. Tim3 expression in Kupffer cell were examined by Real-time PCR and Western-blot. The effects of Tim3 over-expression and Tim3 blocking by anti-Tim3 antibody on mice liver Kupffer cell activation factor （TNF-α, IL-1β and IL-6） were monitored by ELISA test. Jak2/Stat1 proteins were examined by Western blot.Results Real-time PCR detection results show IFN-γ could signiifcantly increase the mice liver Kupffer cells Tim3 mRNA expression levels （P 〈 0.05）. TNF-α, IL-6 and IL-1β of pcDNA3.1-Tim3 group of decreased signiifcantly than the control group （P〈 0.01）; according to the results of ELISA, TNF-α, IL-6, IL-1β of Tim3 blocking antibody group increased signiifcantly than control group （P〈 0.01）. Western blot test showed that compared with the control group, Jak2 and Sat1 protein expression of Tim3 blocking antibodies group increased signiifcantly than control group （P 〈 0.05）. Conclusion Tim3 is involved in the regulation of Kupffer cells activation through Jak2/Stat1 protein.

关 键 词：Tim3 KUPFFER细胞 小鼠 

分 类 号：R392[医药卫生—免疫学]