National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Pathum Thani, Thailand.
Department of Preclinical Science, Faculty of Medicine, Thammasat University, Khlong Luang, Pathum Thani, Thailand
Infect Immun. 2021 May 17;89(6). doi: 10.1128/IAI.00053-21.
infection is the leading cause of chronic gastritis, which can develop into gastric cancer. Eliminating infection with antibiotics achieves the prevention of gastric cancer. Currently, the prevalence of resistance to clarithromycin and metronidazole, and the dual resistance to metronidazole and clarithromycin (C_R, M_R, and C/M_R, respectively), remains at a high level worldwide. As a means of exploring new candidate proteins for the management of infection, secreted proteins from antibiotic-susceptible and antibiotic-resistant -associated gastritis strains were obtained by in-solution tryptic digestion coupled with nano-liquid chromatography tandem mass spectrometry (nano-LC-MS/MS). A total of 583, 582, 590, and 578 differential expressed proteins were identified from C_R, M_R, C/M_R, and antibiotic-sensitive strain (S_S) samples, respectively. Of these, 23 overlapping proteins were found by Venn diagram analysis. Based on heat map analyses, the most and least differing protein expressions were observed from C/M_R strains and S_S strains, respectively. Of the proteins secreted by the S_S strain, only nine were found. After predicting the protein interaction with metronidazole and clarithromycin via the STITCH database, the two most interesting proteins were found to be rpoBC and FBPAII. After quantitative real-time reverse transcription PCR (qRT-PCR) analysis, a downregulation of from M_R strains was observed, suggesting a relationship of to metronidazole sensitivity. Inversely, an upregulation of from C_R, M_R, and C/M_R strains was noticed, suggesting the paradoxical expression of FBPAII and the gene. This report is the first to demonstrate the association of these two novel secreted proteins, namely, rpoBC and FBPAII, with antibiotic-sensitive associated gastritis strains.
感染是慢性胃炎的主要原因，慢性胃炎可能发展为胃癌。用抗生素消除感染可预防胃癌。目前，全球范围内克拉霉素和甲硝唑耐药率以及甲硝唑和克拉霉素双重耐药率（分别为 C_R、M_R 和 C/M_R）居高不下。为了探索用于管理感染的新候选蛋白，通过溶液中胰蛋白酶消化与纳升液相色谱串联质谱（nano-LC-MS/MS）相结合，获得了来自抗生素敏感和抗生素耐药相关胃炎菌株的分泌蛋白。从 C_R、M_R、C/M_R 和抗生素敏感株（S_S）样品中分别鉴定出 583、582、590 和 578 个差异表达蛋白。通过 Venn 图分析发现其中有 23 个重叠蛋白。基于热图分析，C/M_R 株和 S_S 株的蛋白表达差异最大和最小。在 S_S 株分泌的蛋白中，仅发现 9 种。通过 STITCH 数据库预测与甲硝唑和克拉霉素的蛋白相互作用后，发现最有趣的两种蛋白是 rpoBC 和 FBPAII。通过定量实时逆转录 PCR（qRT-PCR）分析，观察到 M_R 株中 表达下调，表明 与甲硝唑敏感性有关。相反，C_R、M_R 和 C/M_R 株中 表达上调，表明 FBPAII 和 基因的表达呈悖论。本报告首次证明了这两种新型分泌蛋白 rpoBC 和 FBPAII 与抗生素敏感相关胃炎菌株之间存在关联。
