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

转运蛋白 HP0939 、 HP0497 和 HP0471 通过增强药物外排参与幽门螺杆菌内在多药耐药和生物膜形成。

  • 影响因子:3.27
  • DOI:10.1111/hel.12715
  • 作者列表:"Cai Y","Wang C","Chen Z","Xu Z","Li H","Li W","Sun Y
  • 发表时间:2020-06-16
Abstract

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.

摘要

背景: 幽门螺杆菌的多药耐药问题日益严重。因此,有必要对幽门螺杆菌多药耐药机制进行研究。我们之前已经确定 HP0939 、 HP0497 和 HP0471 转运蛋白影响幽门螺杆菌药物的流出。由于外排泵参与细菌多药耐药和生物膜形成,我们假设这些转运蛋白可能参与 H.pylori 的多药耐药和生物膜形成。 材料和方法: 因此,我们使用野生型 (WT) 构建了三个基因敲除菌株 Δ hp0939 、 Δ hp0497 和 Δ hp0471,以及三个高表达菌株 Hp0939he 、 Hp0497he 和 Hp0471he 26 695 株 H.pylori 为模板。测定野生株、敲除株、高表达株对阿莫西林、甲硝唑等抗生素的最低抑菌浓度 (MIC) 和最低杀菌浓度 (MBC)。通过 Hoechst 33 342 积累试验比较高表达菌株和野生菌株的外排能力。 结果: 与 WT 相比,敲除菌株对抗生素更敏感,而高表达菌株对抗生素更不敏感。高表达菌株外排药物的能力明显高于 WT。我们还诱导 H.pylori 形成生物膜,观察到与 WT 相比,敲除菌株几乎不能形成生物膜,对几种抗生素更敏感。测定 hp0939 、 hp0497 和 hp0471 在临床敏感和多药耐药菌株中的 mRNA 表达,发现这些基因在临床分离的多药耐药菌株中高表达。 结论: 在本研究中,我们发现三种转运蛋白参与了幽门螺杆菌的内在多药耐药。

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