背景:幽门螺杆菌感染是胃癌的主要病因之一。幽门螺杆菌表达的硫氧还蛋白-1(Trx1)和精氨酸酶(RocF)与其致病性密切相关。然而,Trx1和RocF是否可用于高致病性幽门螺杆菌的临床筛查,以及trx1高表达幽门螺杆菌的发病机制仍然未知。
方法:我们采用逆转录和实时荧光定量PCR(RT-qPCR)检测人胃窦组织中幽门螺杆菌trx1和幽门螺杆菌rocF的表达水平,阐明trx1和rocF在筛选高致病性幽门螺杆菌中的临床应用价值。通过与trx1高表达或低表达幽门螺杆菌共培养的GES-1细胞的RNA-seq,进一步探讨了Trx1的致病机制。通过RT-qPCR验证差异表达基因和信号通路,酶联免疫吸附测定(ELISA),westernblot,免疫组织化学和免疫荧光。我们还评估了trx1高表达和低表达的幽门螺杆菌对GES-1细胞的粘附性。
结果:我们发现H.pyloritrx1和H.pylorirocF在胃癌和消化性溃疡组比胃炎组表达更显著,平行诊断H.pyloritrx1和H.pylorirocF具有较高的敏感性。高表达trx1的H.pylori对GES-1细胞有较强的粘附能力,上调白细胞介素(IL)23A/核因子κB(NF-κB)/IL17A,IL6、IL8通路。
结论:H.pyloritrx1和H.pylorirocF可用于高致病性H.pylori的临床筛查和预测H.pylori感染的结局。trx1高表达H.pylori具有较强的粘附能力,通过上调NF-κB信号通路的激活促进胃部疾病的发生发展。
BACKGROUND: Helicobacter pylori infection is one of the main causes of gastric cancer. thioredoxin-1 (Trx1) and arginase (RocF) expressed by H. pylori were found to be closely related to its pathogenicity. However, whether Trx1 and RocF can be used in clinical screening of highly pathogenic H. pylori and the pathogenesis of trx1 high expressing H. pylori remain still unknown.
METHODS: We investigated the expression level of H. pylori trx1 and H. pylori rocF in human gastric antrum tissues using reverse transcription and quantitative real-time PCR (RT-qPCR) and clarified the clinical application value of trx1 and rocF for screening highly pathogenic H. pylori. The pathogenic mechanism of Trx1 were further explored by RNA-seq of GES-1 cells co-cultured with trx1 high or low expressing H. pylori. Differentially expressed genes and signaling pathways were validated by RT-qPCR, Enzyme-linked immunosorbent assay (ELISA), western blot, immunohistochemistry and immunofluorescence. We also assessed the adherence of trx1 high and low expressing H. pylori to GES-1 cells.
RESULTS: We found that H. pylori trx1 and H. pylori rocF were more significantly expressed in the gastric cancer and peptic ulcer group than that in the gastritis group and the parallel diagnosis of H. pylori trx1 and H. pylori rocF had high sensitivity. The trx1 high expressing H. pylori had stronger adhesion ability to GES-1 cells and upregulated the interleukin (IL) 23A/nuclear factor κappaB (NF-κB)/IL17A, IL6, IL8 pathway.
CONCLUSIONS: H. pylori trx1 and H. pylori rocF can be used in clinical screening of highly pathogenic H. pylori and predicting the outcome of H. pylori infection. The trx1 high expressing H. pylori has stronger adhesion capacity and promotes the development of gastric diseases by upregulating the activation of NF-κB signaling pathway.