伤寒杆菌IVB型菌毛激活PKC、ERK、NF-κB信号通路诱导THP-1细胞IL-6的表达

发布时间：2018-03-25 07:02

本文选题：伤寒杆菌IVB型菌毛　切入点：THP-1细胞　出处：《武汉大学》2005年硕士论文

【摘要】：目的初步探讨伤寒杆菌IVB型菌毛是否参与细菌对机体造成的炎症损伤,是否激活PKC、ERK、NF-κB信号通路诱导THP-1细胞IL-6的表达。方法将THP-1细胞分别与有IVB型菌毛的A21-6菌、缺失IVB型菌毛的pilS菌共同孵育;ELISA法检测IL-6的产量、RT-PCR检测IL-6 mRNA的表达水平、非放射活性测定法检测PKC活性变化、免疫印迹法检测ERK磷酸化水平、荧光素酶报道基因分析法检测NF-κB激活表达。结果 THP-1细胞经有IVB型菌毛的A21-6菌刺激后,IL-6的产生水平、mRNA的表达水平、PKC的活性变化、ERK激酶磷酸化水平和NF-κB激活表达均显著高于相应的pilS菌组;PKC、ERK和NF-κB活化水平分别于刺激10min、30min和60min后即明显升高;并分别于30min、60min和120min左右达到峰值。PKC抑制剂DECA一定程度上能够抑制IVB型菌毛诱导的THP-1细胞PKC活性、NF-κB激活表达和IL-6产生水平。结论伤寒杆菌IVB型菌毛显著增强了THP-1细胞PKC、ERK和NF-κB活化水平,IVB型菌毛在伤寒杆菌诱导单核/巨噬细胞PKC、ERK和NF-κB活化过程中是一种重要的强刺激因素;IVB型菌毛在伤寒杆菌诱导单核/巨噬细胞产生IL-6过程中也是一种重要的刺激因素;PKC、ERK和NF-κB三种信号通路分子参与了该过程的细胞内信号转导。伤寒杆菌IVB型菌毛可能通过激活PKC→ERK→NF-κB途径引起IL-6等致炎因子的大量合成与释放,从而造成机体病理损伤。
[Abstract]:Objective to investigate whether the IVB fimbriae of S. typhimurium are involved in the inflammatory injury caused by bacteria and whether it activates the signal pathway of PKCERKN NF- 魏 B to induce the expression of IL-6 in THP-1 cells. Methods THP-1 cells and A21-6 bacteria with IVB fimbriae were divided into two groups. The expression level of IL-6 mRNA was detected by RT-PCR, PKC activity was detected by non-radioactivity assay, and ERK phosphorylation level was detected by Western blot. Luciferase reporter gene analysis was used to detect the activation and expression of NF- 魏 B. Results THP-1 cells were stimulated by A21-6 strain with IVB type pili. Results the level of IL-6 production and the activity of PKC were correlated with the phosphorylation of ERK kinase and the activation and expression of NF- 魏 B in THP-1 cells. The activation levels of PKC ERK and NF- 魏 B were significantly higher than those in the corresponding pilS group after stimulation for 10 min or 30 min and 60min, respectively. The peak value of PKC inhibitor, DECA, reached a peak at about 30 min or so. PKC inhibitor DECA could inhibit the activation, expression of NF- 魏 B and IL-6 production in THP-1 cells induced by IVB fimbriae to some extent. Conclusion the IVB fimbriae of S. typhimurium can significantly enhance THP-1 cells. The activation level of PKCU ERK and NF- 魏 B the IVB-type fimbriae are an important stimulator in the process of inducing the activation of mononuclear / macrophage PKC- 魏 B by S. typhimurium, and also in the process of inducing monocytes / macrophages to produce IL-6 by S. typhimurium. One of the important stimulant factors, PKC ERK and NF- 魏 B, is involved in the intracellular signal transduction of this process. The IVB fimbriae of S. typhimurium may be activated by the activation of PKC. 鈫扙RK. 鈫扤F- 魏 B pathway induces the synthesis and release of IL-6 and other inflammatory factors, resulting in pathological injury.
【学位授予单位】：武汉大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R378

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