幽门螺杆菌新毒素Tip-α对IL-8、TNF-α、IL-1β表达的影响  

Effects of tumor necrosis factor-α inducing protein on cytokines expression in H.pylori

作　　者：陈王凯[1] 郝波[1] 张国新[1] 施瑞华[1] 程文芳[1] 

机构地区：[1]南京医科大学第一附属医院消化科,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2013年第10期1373-1376,共4页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省高校自然科学研究项目(08KJ3320003)

摘　　要：目的:研究幽门螺杆菌(H.pylori,HP)毒素肿瘤坏死因子α诱导蛋白(Tip-α)对细胞因子白介素(IL)-8、肿瘤坏死因子(TNF)-α、IL-1β表达的影响及其意义。方法:从HP的26695菌株扩增位于H.pylori0596的Tip-α基因,重组载体转化至大肠杆菌表达并纯化Tip-α蛋白,用Western blot方法进行鉴定。以不同浓度的Tip-α作用于胃上皮细胞GES-1,用ELISA方法检测不同时间点GES-1细胞中IL-8、TNF-α、IL-1β表达变化。以四氢吡咯烷(PDTC,NF-κB特异性的抑制剂)阻断NF-κB信号通路后,检测Tip-α作用GES-1后IL-8、TNF-α、IL-1β的表达变化。结果:用Western blot检测纯化蛋白,抗Tip-α抗体可与纯化蛋白特异性结合。Tip-α刺激细胞后细胞因子IL-8、TNF-α、IL-1β的表达水平较刺激前明显升高,刺激前后差别具有统计学意义(P<0.05)。PDTC作用后再给予Tip-α刺激,细胞中IL-8、TNF-α、IL-1β的表达水平与无PDTC作用组相比明显下降(P<0.05)。结论:Tip-α可使细胞因子IL-8、TNF-α、IL-1β表达明显升高,在HP致炎中起重要作用,这种作用可能通过NF-κB途径。Objective：To investigate the effects of new tumor necrosis factor-α-inducing protein （Tip-α） on the expression of cytokines and its significance. Methods：Tip-α were amplified from genome sequence of Helicobacter pylori （H.pylori） strain 26695. The recombinant plasmid was transformed into E.coli. The expression product was purified and confirmed by Western blot. After treated by different concentrations of Tip-α in different times, the level of cytokines （IL-8 ,TNF-α, IL-1 β） in GES-1 were detected by ELISA. They were also detected after NF-KB was inhibited by pyrrolidine dithioearbamate （PDTC,the special inhibitor of NF-KB）. Results：It was confirmed by Western blot analysis that Tip-α antibody combined with purified proteins specifically. When the cells were treated with Tip-or,the levels of cytokines significant increased （P 〈 0.05）. After NF-KB was inhibited by PDTC,the levels of cytokines significant decreased （P 〈 0.05）. Conclusion： Tip-α can promote the expression of cytokines （IL-8, TNF-α, IL- 1β）, it may play an important role in the phlogogenic effect of HP, this effect may be NF-KB signal pathway dependent.

关 键 词：幽门螺杆菌 Tip-α 白介素-8 白介素-1Β 肿瘤坏死因子-α 核因子-KB 

分 类 号：R573.3[医药卫生—消化系统]