Bioinformatics analysis of small RNAs in Helicobacter pylori and the role of NAT-67 under tinidazole treatment

Abstract

Helicobacter pylori (Hp) infection is a major cause of gastrointestinal disease. However, the pathogenesis of gastric mucosa injury by Hp has remained elusive. Small non-coding RNA (sRNA) is a type of widespread RNA in prokaryotic organisms and regulates bacterial growth, reproduction and virulence. In the present study, Hp sRNA profiles were generated to reveal the sequences and possible functions of sRNA by bioinformatics analysis. The role of sRNA in tinidazole (TNZ) treatment was also explored. Total sRNA s of HP26695 were sequenced using an Illumina HiSeq2000. Detected Tags were then compared with a known sRNA database to build an sRNA profile. Reverse transcription-quantitative (RT-q) PCR products were sequenced directly and agarose gel electrophoresis was used to identify NA T-67 and 5'ureB-sRNA in HP. Furthermore, HP was treated with TNZ for 6, 12 and 24 h. The bacterial concentration was measured, the expression of NA T-67, 5'ureB-sRNA and ceuE was determined by RT-qPCR and superoxide dismutase (SOD) activity and reactive oxygen species (ROS) production were detected. A total of 163 sRNA tags were predicted in Hp through bioinformatics analysis. Among them, 35 tags were evolutionarily aconserved in different Hp strains. By target prediction, it was indicated that certain candidate sRNA s were associated with bacterial oxidative stress, virulence and chemotaxis. It was also observed that NA T-67 and 5'ureB-sRNA were downregulated in TNZ-treated HP. TNZ treatment inhibited the growth of Hp, which was accompanied by downregulation of ceuE and SOD activity, as well as upregulation of RO S. RNA sequencing and bioinformatics are valuable in predicting the expression profile and function of sRNA in HP. sRNA- targeted genes may be associated with virulence, oxidative stress and chemokines. Downregulation of NA T-67 by TNZ may be involved in Hp oxidative stress regulation, which may comprise one of the mechanisms of the antibacterial effects of TNZ.