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

幽门螺杆菌新毒素Tip-α对IL-8、TNF-α、IL-1β表达的影响 陈王凯　郝波　 张国新 施瑞华?基金项目：江苏省高校自然科学研究 ?基金项目：江苏省高校自然科学研究项目（项目批准号：08KJ3320003） 作者单位：210029 南京医科大学第一附属医院消化科 通讯作者：程文芳，chengwenfang@ 【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(P0.05)。PDTC作用后，再给予Tip-α刺激细胞中IL-8、TNF-α、IL-1β的表达水平与无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，在HP致炎中起重要作用，这种作用可能通过NF-κB途径。 【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-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 HYPERLINK dict://key.0895DFE8DB67F9409DB285590D870EDD/significance \o 查找: significance 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