论文部分内容阅读
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 effect of Tip-α of Helicobacter pylori (H.pylori) on the expression of interleukin (IL) -8, tumor necrosis factor (TNF) -α and IL-1β And its significance. Methods: The Tip-α gene of H.pylori0596 was amplified from strain 26695 of HP. The recombinant vector was transformed into E.coli and the Tip-α protein was purified and identified by Western blot. The effects of different concentrations of Tip-α on GES-1 in gastric epithelial cells were detected by ELISA. The expressions of IL-8, TNF-α and IL-1β in GES-1 cells were detected by ELISA. The expression of IL-8, TNF-α and IL-1β after Tip-α treatment of GES-1 was detected by blocking the NF-κB signaling pathway with tetrahydropyrrolidine (PDTC, a specific inhibitor of NF-κB) Results: The purified protein was detected by Western blot. The anti-Tip-α antibody could specifically bind with the purified protein. The expression of IL-8, TNF-α and IL-1β in the cells stimulated by Tip-α was significantly higher than that before stimulation, and the difference was statistically significant (P <0.05). After the treatment with PDTC, the expression of IL-8, TNF-α and IL-1β in the cells was significantly decreased compared with the control group without PDTC (P <0.05). Conclusion: Tip-α can significantly increase the expression of cytokines IL-8, TNF-α and IL-1β and play an important role in the inflammation of HP. This effect may be through the NF-κB pathway.