中国免疫学杂志
中國免疫學雜誌
중국면역학잡지
CHINESE JOURNAL OF IMMUNOLOGY
2014年
10期
1313-1319
,共7页
涂文娟%汪小庭%刘亮明%朱彤%梁冬雨%杨志文%高得勇
塗文娟%汪小庭%劉亮明%硃彤%樑鼕雨%楊誌文%高得勇
도문연%왕소정%류량명%주동%량동우%양지문%고득용
枯否细胞%炎症%脂多糖%UrotensinⅡ%Urantide%Toll样受体4%干扰素调节因子3
枯否細胞%炎癥%脂多糖%UrotensinⅡ%Urantide%Toll樣受體4%榦擾素調節因子3
고부세포%염증%지다당%UrotensinⅡ%Urantide%Toll양수체4%간우소조절인자3
Kupffer cell%Inflammation%Lipopolysaccharide%Urotensin Ⅱ%Urantide%TLR4%IRF3
目的:探讨Urotensin Ⅱ( 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。
目的:探討Urotensin Ⅱ( UⅡ)/UT繫統對脂多糖( Lipopolysaccharide ,LPS)刺激枯否細胞( Kupffer cell ,KC)固有免疫炎癥信號通路Toll樣受體4(Toll-like receptor 4,TLR4)-榦擾素調節因子3(Interferon regulatory factor 3,IRF3)的影響。方法:體外分離培養大鼠肝KCs,KCs培養上清液促炎性細胞因子IL-6、IFN-β和IFN-γ分泌水平採用ELISA分析檢測,細胞錶麵TLR4的錶達採用流式細胞技術分析,IRF3基因和蛋白錶達情況分彆採用real-time PCR和Western blot 分析方法檢測。結果:LPS刺激後,KCs培養上清液IL-6、IFN-β和IFN-γ分泌水平、細胞錶麵TLR4錶達暘性細胞率及細胞內IRF3 mRNA錶達水平均顯著升高,UT拮抗劑urantide預處理抑製瞭LPS刺激誘導KCs對上述分子的上調錶達;LPS的應用也造成瞭KCs胞覈內IRF3蛋白錶達水平升高,而使胞漿內IRF3蛋白錶達水平降低,urantide預處理後,抑製瞭LPS誘導KCs覈內IRF3蛋白上調和胞漿水平下調。結論:UⅡ/UT繫統通過對TLR4-IRF3通路的正性調控作用,介導瞭或至少部分介導瞭LPS刺激KCs的免疫性炎癥分泌效應。
목적:탐토Urotensin Ⅱ( UⅡ)/UT계통대지다당( Lipopolysaccharide ,LPS)자격고부세포( Kupffer cell ,KC)고유면역염증신호통로Toll양수체4(Toll-like receptor 4,TLR4)-간우소조절인자3(Interferon regulatory factor 3,IRF3)적영향。방법:체외분리배양대서간KCs,KCs배양상청액촉염성세포인자IL-6、IFN-β화IFN-γ분비수평채용ELISA분석검측,세포표면TLR4적표체채용류식세포기술분석,IRF3기인화단백표체정황분별채용real-time PCR화Western blot 분석방법검측。결과:LPS자격후,KCs배양상청액IL-6、IFN-β화IFN-γ분비수평、세포표면TLR4표체양성세포솔급세포내IRF3 mRNA표체수평균현저승고,UT길항제urantide예처리억제료LPS자격유도KCs대상술분자적상조표체;LPS적응용야조성료KCs포핵내IRF3단백표체수평승고,이사포장내IRF3단백표체수평강저,urantide예처리후,억제료LPS유도KCs핵내IRF3단백상조화포장수평하조。결론:UⅡ/UT계통통과대TLR4-IRF3통로적정성조공작용,개도료혹지소부분개도료LPS자격KCs적면역성염증분비효응。
Objecitve: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-in-flammatory cytokines including IL-6,IFN-βand IFN-γwere 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-βand IFN-γsecretion, 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 TLR 4-IRF3 pathway in LPS-stimulated KCs.