中华实验和临床感染病杂志（电子版）
中華實驗和臨床感染病雜誌（電子版）
중화실험화림상감염병잡지（전자판）
Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Version)
2015年
5期
703-707
,共5页
牛坚%王月%李克清%朱志军%刘斌
牛堅%王月%李剋清%硃誌軍%劉斌
우견%왕월%리극청%주지군%류빈
Tim3%Kupffer细胞%小鼠
Tim3%Kupffer細胞%小鼠
Tim3%Kupffer세포%소서
T cell immune globulin and mucin family-3%Kupffer cell%Mice
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pcDNA3.1-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pcDNA3.1-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 mRNA的表达水平(P< 0.05);pcDNA3.1-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.
目的 探討T細胞免疫毬蛋白及黏蛋白傢族-3(Tim3)對IFN-γ活化的小鼠Kupffer細胞的調節作用併探討其相關機製.方法 將真覈錶達質粒pcDNA3.1-Tim3轉染小鼠肝Kupffer細胞,以Real-time PCR和Western blot檢測Tim3在小鼠肝Kupffer細胞的錶達.通過ELISA檢測質粒pcDNA3.1-Tim3、Tim3阻斷型抗體對IFN-γ活化的小鼠肝Kupffer細胞因子(TNF-α、IL-1β和IL-6)產生的影響,Western blot檢測JAK2/STATl蛋白錶達.結果 Real-time PCR檢測結果顯示,IFN-γ能夠顯著提高小鼠肝髒Kupffer細胞中Tim3 mRNA的錶達水平(P< 0.05);pcDNA3.1-Tim3組的TNF-α、IL-6和IL-1β分泌較對照組顯著下降(P< 0.01);ELISA結果顯示,Tim3阻斷型抗體組與對照組相比,TNF-α、IL-6和IL-1β的分泌增加(P< 0.01);Western blot檢測顯示,與對照組相比,Tim3阻斷型抗體預處理的小鼠Kupffer細胞JAK2及STAT1蛋白的錶達上調(P< 0.05).結論 Tim3通過調控Jak2/Stat1蛋白錶達參與瞭Kupffer細胞活化的調節.
목적 탐토T세포면역구단백급점단백가족-3(Tim3)대IFN-γ활화적소서Kupffer세포적조절작용병탐토기상관궤제.방법 장진핵표체질립pcDNA3.1-Tim3전염소서간Kupffer세포,이Real-time PCR화Western blot검측Tim3재소서간Kupffer세포적표체.통과ELISA검측질립pcDNA3.1-Tim3、Tim3조단형항체대IFN-γ활화적소서간Kupffer세포인자(TNF-α、IL-1β화IL-6)산생적영향,Western blot검측JAK2/STATl단백표체.결과 Real-time PCR검측결과현시,IFN-γ능구현저제고소서간장Kupffer세포중Tim3 mRNA적표체수평(P< 0.05);pcDNA3.1-Tim3조적TNF-α、IL-6화IL-1β분비교대조조현저하강(P< 0.01);ELISA결과현시,Tim3조단형항체조여대조조상비,TNF-α、IL-6화IL-1β적분비증가(P< 0.01);Western blot검측현시,여대조조상비,Tim3조단형항체예처리적소서Kupffer세포JAK2급STAT1단백적표체상조(P< 0.05).결론 Tim3통과조공Jak2/Stat1단백표체삼여료Kupffer세포활화적조절.
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.
