幽门螺杆菌新毒素Tip-α对IL-8、TNF-α、IL-1β表达影响.docVIP

幽门螺杆菌毒素Tip-α对IL-8、TNF-α、IL-1β表达的影响　郝波　 张国新 施瑞华(程文芳 【摘要】目的 研究幽门螺杆菌H.pylori，HP）毒素Tip-α(肿瘤坏死因子α诱导蛋白)对细胞因子IL-8、TNF-α、IL-1β表达的影响及其。方法 从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GES-1），ELISA方法检测不同时间GES-1细胞中细胞因子IL-8、TNF-α、IL-1β表达变化。以PDTC四氢吡咯烷NF-κB特异性的抑制剂阻断NF-κB后检测Tip-α作用GES-1后IL-8、TNF-α、IL-1β的变化。结果 用estern blot检测纯化蛋白，抗Tip-α抗体纯化蛋白。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(P0.05)。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PDTC作用组相比明显下降(P0.05)。结论 Tip-α可使细胞因子IL-8、TNF-α、IL-1β表达明显升高，NF-κB途径。 【关键词】幽门螺杆菌；Tip-α；；-1β；肿瘤坏死因子-α；核因子-κB Tumor necrosis factor-α inducing protein of H.pylori effects on cytokines expression CHEN Wang-kai, HAO Bo, ZHANG Guoxin, SHI Rui-hua， CHENG Wen-fang,. Department of Gastroenterology, The First Hospital Affiliated to Nanjing Medical University, Nanjing 210029,CHINA Corresponding author：CHENG Wen-fang，Email：chengwenfang@ 【Abstact】objective To investigate the effects of Tip-α (New tumor necrosis factor-a-inducing protein)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 confirmed by western blotting. By different concentrations of Tip-α affectting GES-1 in different times, the level of cytokines（IL-8、TNF-α、IL-1β ）were detected by ELISA. The level of them was　also detected in GES-1 after NF-κB was inhibited by PDTC (Pyrrolidine Dithioearbamate, the special inhibitor of NF-κB). Results Tip-α antibody specificity combine with purified proteins by Western blot analysis. When Tip-α interfered the cells ,the level of cytokines was significant higher than that of control(P 0.05).. After cutting off the NF-κB by PDTC, we found that the level of cytokines were significant lower than that of only Tip-α interfering (P0.05). Conclusion Tip-α can promote the expression of cytokines（ IL-8、TNF-α、IL-1βplay an important role