Transporters HP0939, HP0497, and HP0471 participate in intrinsic multidrug resistance and biofilm formation in Helicobacter pylori by enhancing drug efflux

• Published in: Helicobacter (Cambridge, Mass.) Mass.), 2020-08, Vol.25 (4), p.e12715-n/a
• Main Authors: Cai, Yuying, Wang, Caixia, Chen, Zhenghong, Xu, Zhengzheng, Li, Huanjie, Li, Wenjuan, Sun, Yundong
• Format: Article
• Language: English
• Subjects: Amoxicillin; Anti-Bacterial Agents - metabolism; Anti-Bacterial Agents - pharmacology; Antibiotics; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bile; biofilm; Biofilms; Biofilms - growth & development; Drug Resistance, Multiple, Bacterial - drug effects; Drug Resistance, Multiple, Bacterial - genetics; Efflux; efflux pump; Gene Expression; H pylori; Helicobacter Infections - microbiology; Helicobacter pylori; Helicobacter pylori - drug effects; Helicobacter pylori - genetics; Helicobacter pylori - isolation & purification; Helicobacter pylori - physiology; Humans; Membrane Transport Proteins - genetics; Membrane Transport Proteins - metabolism; Metronidazole; Microbial Sensitivity Tests; Minimum inhibitory concentration; Multidrug resistance; Multidrug resistant organisms; Strains (organisms); transporter
• ISSN: 1083-4389; 1523-5378
• DOI: 10.1111/hel.12715

Background The multidrug resistance of Helicobacter pylori is becoming an increasingly serious issue. It is therefore necessary to study the mechanism of multidrug resistance of H pylori. We have previously identified that the HP0939, HP0497, and HP0471 transporters affect the efflux of drugs from H pylori. As efflux pumps participate in bacterial multidrug resistance and biofilm formation, we hypothesized that these transporters could be involved in the multidrug resistance and biofilm formation of H pylori. Materials and Methods We therefore constructed three knockout strains, Δhp0939, Δhp0497, and Δhp0471, and three high‐expression strains, Hp0939he, Hp0497he, and Hp0471he, using the wild‐type (WT) 26 695 strain of H pylori as the template. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of wild strains, knockout strains, and high‐expression strains to amoxicillin, metronidazole, and other antibiotics were measured. The efflux capacity of high‐expression strains and wild strains was compared by Hoechst 33 342 accumulation assay. Results Determination of the MIC and MBC of the antibiotics revealed that the knockout strains were more sensitive to antibiotics, while the high‐expression strains were less sensitive to antibiotics, compared to the WT. The ability of the high‐expression strains to efflux drugs was significantly higher than that of the WT. We also induced H pylori to form biofilms, and observed that the knockout strains could barely form biofilms and were more sensitive to several antibiotics, compared to the WT. The mRNA expression of hp0939, hp0497, and hp0471 in the clinically sensitive and multidrug‐resistant strains was determined, and it was found that these genes were highly expressed in the multidrug‐resistant strains that were isolated from the clinics. Conclusions In this study, we found three transporters involved in intrinsic multidrug resistance of H pylori.