CAJ | 학술논문

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。
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.