NF-κB信号通路对BLP耐受巨噬细胞iNOS表达的影响  

The effect of NF-κB signaling pathway on iNOS expression in BLP-tolerized macrophages

作　　者：李雪[1] 王义乾[1] 罗海华[1] 钟玙沄[1] 雷烨铭 雷山 蔡军伟[1] 姜勇[1] 刘靖华[1] 

机构地区：[1]广东省蛋白质组学重点实验室,南方医科大学病理生理学教研室,广州510515

出　　处：《中国临床解剖学杂志》2014年第3期300-305,共6页Chinese Journal of Clinical Anatomy

基　　金：国家自然科学基金(81272149;81072425)

摘　　要：目的通过检测细菌脂蛋白(BLP)耐受巨噬细胞感染细菌时诱导型一氧化氮合酶(iNOS)表达情况及其表达是否受NF-κB信号通路调控,探讨BLP耐受巨噬细胞对细菌清除能力增强的机制。方法比较BLP耐受和非耐受(Naive)小鼠骨髓来源的巨噬细胞(BMM)对大肠杆菌的吞噬及杀灭情况,评价BLP耐受巨噬细胞的细菌清除能力;用定量PCR技术检测BLP耐受的BMM内iNOS mRNA表达情况和细胞免疫荧光技术观察p65从胞质向胞核的移位情况;最后观察抑制NF-κB通路活化对iNOS mRNA表达的影响。结果 BLP耐受巨噬细胞吞噬细菌和杀灭细菌的能力较Naive细胞显著增强(P<0.05);iNOS mRNA表达水平较Naive细胞显著(P<0.05);如果抑制BLP耐受巨噬细胞NF-κB通路活化对iNOS mRNA表达有显著影响(P<0.05)。结论本研究结果提示细菌脂蛋白耐受通过NF-κB通路活化增强细菌感染巨噬细胞iNOS表达。Objective In order to explore mechanisms underlying enhanced bacterial clearance of BLP-tolerized macrophages,we detected the expression of inducible nitric oxide synthase （iNOS） and investigated whether this expression was regulated by NF- κB signaling pathway in BLP- tolerized macrophages to bacterial infection. Methods Through comparison of the phagocytosis and intracellular bacterial killing of E.coli between Naive and BLP-tolerized mice bone marrow-derived macrophages （BMMs）, we evaluated the bacterial clearing capability of BLP-tolerized macrophages. Next, the mRNA level of iNOS in BLP-tolerized macrophages was detected by real-time PCR, and the translocations of p65 from cytoplasm to nucleus were shown by immunofluorescence.Finally, the activation of NF-κB signaling pathway was inhibited and the mRNA expression of iNOS in BLP-tolerized BMMs were observed. Results Compared to Naive macrophages, the phagocytosis and intracellular bacterial killing of BLP-tolerized macrophages were significantly enhanced （P〈0.05）. The mRNA level of iNOS in BLP-tolerized macrophages was significantly increased （P〈0.05）, which could be significantly affected when the activation of NF-κB signaling pathway was inhibited （P〈0.05）. Conclusion The study suggests that iNOS expression increases through the activation of NF-κB signaling pathway in BLP-tolerized macrophages to bacterial infection.

关 键 词：BLP耐受 细菌吞噬 INOS NF—κB 

分 类 号：R364.5[医药卫生—病理学] R37[医药卫生—基础医学]