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目的研究幽门螺杆菌(Helicobacter pylori,H.pylori)感染对NLRP3炎症小体相关蛋白的活化作用,探讨NLRP3炎症小体在H.pylori感染中的作用。方法培养H.pylori国际标准株NCTC11639,感染胃黏膜上皮细胞株GES-1和胃癌细胞株MKN-45,细菌数∶细胞数(感染复数)为100∶1和200∶1,于6 h和12 h收集细胞,提取总蛋白。Western blotting检测炎症小体中NLRP3及Caspase-1蛋白的表达水平,ELISA法检测细胞中IL-1β的含量。结果 H.pylori感染GES-1细胞后,NLRP3、Caspase-1以及IL-1β的表达随着感染复数的增加而增加,差异均具有统计学意义(P<0.05);感染MKN-45细胞后,NLRP3的蛋白表达随着感染复数的增加而增加,而Caspase-1及IL-1β的表达随着感染复数的增加而降低,差异均有统计学意义(P<0.05);相同感染复数的H.pylori感染GES-1细胞和MKN-45细胞,6 h和12 h之间NLRP3、Caspase-1以及IL-1β的表达差异均无统计学意义(P>0.05)。结论 H.pylori感染在体外能活化GES-1细胞中NLRP3炎症小体相关蛋白NLRP3和Caspase-1,活化MKN-45细胞中NLRP3蛋白的表达,抑制Caspase-1蛋白的表达;促进上皮细胞GES-1中IL-1β炎症因子的表达,抑制肿瘤细胞MKN-45中IL-1β炎症因子的表达。
Objective To investigate the activation of NLRP3 inflammasome-associated proteins by Helicobacter pylori (H.pylori) infection and to explore the role of NLRP3 inflammasome in H.pylori infection. Methods The gastric mucosal epithelial cell line (GES-1) and gastric cancer cell line MKN-45 were infected by H.pylori international standard strain NCTC11639. The number of bacteria: the number of cells with multiplicity of infection was 100:1 and 200:1, h Collect cells and extract total protein. Western blotting was used to detect the expression of NLRP3 and Caspase-1 in inflammatory corpuscle and the content of IL-1β in the cells by ELISA. Results The expression of NLRP3, Caspase-1 and IL-1β in H.pylori-infected GES-1 cells increased with the increase of the multiplicity of infection, the difference was statistically significant (P <0.05). After infection of MKN-45 cells, The protein expression of NLRP3 increased with the increase of the multiplicity of infection, while the expression of Caspase-1 and IL-1β decreased with the increase of the number of infection, the difference was statistically significant (P <0.05); There was no significant difference in the expression of NLRP3, Caspase-1 and IL-1β between 6 and 12 h after pylori infection of GES-1 cells and MKN-45 cells (P> 0.05). Conclusions H. pylori infection can activate NLRP3 inflammasome related proteins NLRP3 and Caspase-1, activate NLRP3 protein expression in MKN-45 cells and inhibit the expression of Caspase-1 protein in GES-1 cells in vitro, and promote epithelial cell GES- 1 in the expression of IL-1βinflammatory cytokines, inhibiting the expression of IL-1βin inflammatory cytokines in tumor cells MKN-45.