金黄葡萄球菌杀白细胞素（PVL）对THP-1巨噬细胞IL-8、IL-6表达的影响  

Panton-valentine leukocidin induce THP-1 macrophages IL-8 and IL-6 response through nuclearTLR4/NF-KB pathway

作　　者：周馨[1] 马筱玲[1] 常文娇[1] 张翠萍[1] 谢强[1] 

机构地区：[1]安徽医科大学附属省立医院检验科,合肥230001

出　　处：《中华微生物学和免疫学杂志》2011年第10期875-880,共6页Chinese Journal of Microbiology and Immunology

基　　金：安徽省自然科学基金（11040606M205）

摘　　要：目的研究金黄葡萄球菌杀白细胞素（panton-valentineleucocidin，PVL）对THP．1巨噬细胞Toll样受体4（TLR4）／核因子KB（NF-KB）信号通路及IL-8、IL-6的表达影响，探讨PVL相关肺组织损伤的致病机制。方法实验前用100nmol／L佛波酯（PMA）孵育THP-1细胞48h，使其诱导分化成巨噬细胞，同体外诱导PVL重组表达载体表达纯化的LukS-PVI。和LukF．PVL组分共同孵育，采用ELISA法检测细胞上清IL-8、IL-6蛋白的表达，RT-PCR法检测TLR4、IL-8和IL-6mRNA水平表达，免疫组化和Westernblot检测NF-KB的移位及其表达。结果rPVL能促进THP-1巨噬细胞分泌IL-6和IL-8，而且具有明显的浓度和时间依赖性。同时，rPVL能上调TLR4表达，活化NF．KB蛋白。结论PVL相关肺组织损伤机制可能与PVL激活TLR4／NF-KB信号通路，启动炎性因子大量释放有关。Objective To investigate the influence of panton-valentine leucocidin （PVL） on ex- pression of Toll like receptor 4 （TLR4）/nuclear factor-kappa B （ NF-KB ） signals and IL-8, IL-6 in THP-1 macrophages, and to study the mechanism of PVL-related lung tissue damage. Methods THP-1 cells were cuhured in the presence of 100 nmol/L phorbol-12-myristate 3-acetate （PMA） for 48 h to induce monocytemacrophage differentiation, rPVL-F and rPVL-S were induced and expressed from the recombinant plasmid, respectively purified with chromatographic column. After that, THP-1 macrophages were incubated with rPVL, and then ELISA was performed to test expression of IL-8 and L-6 in supernatants fluid ; RT-PCR was performed to detect expression of IL-8, L-6 and TLR4 ; NF-KB was analyzed by Western blot and immunohistochemistry method. Results PVL was able to induce expression of IL-8 and IL-6 in THP-1 macrophages in time-and concentration-dependent manners. PVL could also significantly promote the activation of TLR4/NF- KB signals. Conclusion PVL can activate the expression of TLR4/NF-KB signals, and increased the high expression of inflammatory cytokines. Maybe it＇s the mechanism of action of PVL exerts the function of lung tissue damage.

关 键 词：杀白细胞素 THP-1巨噬细胞 核因子.KB IL-8 IL-6 

分 类 号：R[医药卫生]