NF-κB信号通路对BLP耐受巨噬细胞iNOS表达的影响
收稿日期: 2014-03-26
网络出版日期: 2014-06-04
基金资助
国家自然科学基金（81272149, 81072425）
The effect of NF-κB signaling pathway on iNOS expression in BLP-tolerized macrophages
Received date: 2014-03-26
Online published: 2014-06-04
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。
李雪 , 王义乾 , 罗海华 , 钟玙沄 , 雷烨铭 , 雷山 , 蔡军伟 , 姜勇 , 刘靖华 . NF-κB信号通路对BLP耐受巨噬细胞iNOS表达的影响[J]. 中国临床解剖学杂志, 2014 , 32(3) : 300 -305 . DOI: 10.13418/j.issn.1001-165x.2014
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.
Key words： BLP tolerance; Bacterial phagocytosis; iNOS; NF-κB
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