孙锦霞,黄 钟,王 瑞,等.巨噬细胞中FBW7沉默对其杀菌能力的抑制[J].深圳大学学报理工版,2017,34(No.4(331-440)):358-357.[doi:10.3724/SP.J.1249.2017.04358]
Sun Jinxia,Huang Zhong,Wang Rui,et al.The anti-bactericidal effects of FBW7 silence in macrophage[J].Journal of Shenzhen University Science and Engineering,2017,34(No.4(331-440)):358-357.[doi:10.3724/SP.J.1249.2017.04358]
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巨噬细胞中FBW7沉默对其杀菌能力的抑制
孙锦霞1,黄 钟1,王 瑞2,翁林军3,李亚旭3

1)深圳大学医学部,广东深圳518060; 2)聊城大学药学院,山东聊城252059; 3)同济大学生命科学与技术学院,上海200092

小干扰核糖核酸; RAW264.7细胞; 巨噬细胞; 杀菌活性; 一氧化氮合成酶; NF-κB信号通路; 实时荧光定量聚合酶链式反应

The anti-bactericidal effects of FBW7 silence in macrophage
Sun Jinxia1, Huang Zhong1, Wang Rui2, Weng Linjun3, and Li Yaxu3

Sun Jinxia1, Huang Zhong1, Wang Rui2, Weng Linjun3, and Li Yaxu31)Health Science Center, Shenzhen University, Shenzhen 518060, Guangdong Province, P.R.China2)College of Pharmacy, Liaocheng University, Liaocheng 252059, Shandong Province, P.R.China3)School of Life Science and Technology, Tongji University, Shanghai 200092, P.R.China

small interfering RNA(siRNA); RAW264.7; macrophage; bactericidal activity; nitric oxide synthase(iNOS); NF-κB signaling pathway; real time quantitative polymerase chain reaction(RT-qPCR)

DOI: 10.3724/SP.J.1249.2017.04358

备注

采用小干扰核糖核酸(siRNA)介导Hela细胞、原代巨噬细胞和RAW264.7细胞中FBW7基因沉默,研究其对NF-κB信号通路、细胞因子表达、巨噬细胞吞噬和杀伤细菌能力的影响.通过流式细胞术、抗生素保护试验、实时荧光定量聚合酶链式反应和报告基因的研究方法,发现FBW7沉默不具有调控RAW264.7细胞吞噬细菌的能力,但能显著抑制原代巨噬细胞和RAW264.7细胞的杀菌能力及一氧化氮合成酶(nitric oxide synthase, iNOS)表达,抑制Hela中NF-κB报告基因活性,降低Hela细胞中TNFα、 IL-1β和IL-6, 以及原代巨噬细胞、RAW264.7细胞中MCP-1和IL-1β表达.结果表明,FBW7沉默可通过影响NF-κB活性抑制巨噬细胞对细菌的杀伤能力.

SiRNA-mediated FBW7 silence in Hela, primary macrophage and RAW264.7 were performed to assay the role of FBW7 on NF-κB activity, cytokines expression and phagocytosis and bactericidal activity of macrophage. Flow cytometry(FCM), antibiotic protection assay(APA), real time quantitative polymerase chain reaction(RT-qPCR)and report gene analysis suggest that FBW7 silence does not affect phagocytosis of RAW264.7, but significantly inhibits bactericidal activity and nitric oxide synthase(iNOS)expression in primary macrophage and RAW264.7, suppresses NF-κB report gene activity in Hela, decreases expression of TNFα、 IL-1β、 IL-6 in Hela and MCP-1、 IL-1β in primary macrophage and RAW264.7. Thus, FBW7 silence might inhibit bactericidal activity of macrophage by regulating NF-κB activity.

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