UⅡ/UT系统对LPS刺激枯否细胞固有免疫炎症信号通路TLR4-IRF3的影响  

Roles of UⅡ /UT system played in innate immune inflammatory signal pathway TLR4-IRF3 in LPS-stimulated primary Kupffer cells

作　　者：涂文娟[1,2] 汪小庭 刘亮明[1,2] 朱彤[1,2] 梁冬雨[1,2] 杨志文[1,2] 高得勇[1,2] 

机构地区：[1]南京医科大学附属上海松江中心医院 [2]上海交通大学附属第一人民医院松江分院感染科,上海201600

出　　处：《中国免疫学杂志》2014年第10期1313-1319,共7页Chinese Journal of Immunology

基　　金：国家自然科学基金项目(81070357;30660066)

摘　　要：目的:探讨UrotensinⅡ(UⅡ)/UT系统对脂多糖(Lipopolysaccharide,LPS)刺激枯否细胞(Kupffer cell,KC)固有免疫炎症信号通路Toll样受体4(Toll-like receptor 4,TLR4)-干扰素调节因子3(Interferon regulatory factor 3,IRF3)的影响。方法:体外分离培养大鼠肝KCs,KCs培养上清液促炎性细胞因子IL-6、IFN-β和IFN-γ分泌水平采用ELISA分析检测,细胞表面TLR4的表达采用流式细胞技术分析,IRF3基因和蛋白表达情况分别采用real-time PCR和Western blot分析方法检测。结果:LPS刺激后,KCs培养上清液IL-6、IFN-β和IFN-γ分泌水平、细胞表面TLR4表达阳性细胞率及细胞内IRF3 mRNA表达水平均显著升高,UT拮抗剂urantide预处理抑制了LPS刺激诱导KCs对上述分子的上调表达;LPS的应用也造成了KCs胞核内IRF3蛋白表达水平升高,而使胞浆内IRF3蛋白表达水平降低,urantide预处理后,抑制了LPS诱导KCs核内IRF3蛋白上调和胞浆水平下调。结论:UⅡ/UT系统通过对TLR4-IRF3通路的正性调控作用,介导了或至少部分介导了LPS刺激KCs的免疫性炎症分泌效应。Objective：To investigate effects of urotensin Ⅱ（UⅡ）/UT system on innate immune inflammatory signal pathway TLR4-IRF3 in the lipopolysaccharide（LPS）-stimulated Kupffer cells（ KCs）. Methods： Rat KCs were isolated and cultured. Pro-inflammatory cytokines including IL-6,IFN-β and IFN-γ were assayed by ELISA in culture supernatant of KCs. Cell surface TLR4 were tested with flow cytometry technique. Expression of IRF3 were tested with real-time PCR and Western blot. Results： Significant increases were showed in IL-6,IFN-β and IFN-γ secretion,TLR4-expressed positive rates and IRF3 mRNA levels in KCs after stimulated by LPS,but were inhibited via urantide pretreatment. In addition,LPS induced upregulation of nuclear IRF3 protein and downregulation of cytoplasm IRF3 protein in KCs,which were blocked by urantide pretreatment. Conclusion： UⅡ /UT system mediates immune inflammatory response in part through activating TLR4-IRF3 pathway in LPS-stimulated KCs.

关 键 词：枯否细胞 炎症 脂多糖 UrotensinⅡ URANTIDE Toll样受体4 干扰素调节因子3 

分 类 号：R575.3[医药卫生—消化系统]