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目的 体外探究盐酸小檗碱对幽门螺杆菌（H. pylori）及其生物膜脲酶基因ureA和ureB表达的影响。方法 复苏本实验室保存的H. pylori的标准菌株26695和4株临床分离来源的H. pylori耐药株H390、e660、H57、H389；通过肉汤稀释法测定盐酸小檗碱对上述5株H. pylori的浮游菌的最低抑菌浓度（MIC）；通过棋盘格法测定盐酸小檗碱与阿莫西林对上述4株临床耐药的H. pylori的联合抑菌作用；通过肉汤稀释法测定盐酸小檗碱对26695和H390菌株的生物膜的MIC；将0.5MIC、MIC、2MIC盐酸小檗碱浓度分别与上述26695和H390菌株的H. pylori的浮游菌及生物膜共孵育2、8、24 h，对其脲酶活性进行检测，并通过RT-qPCR检测ureA和ureB基因的表达情况。结果 盐酸小檗碱对抗生素敏感菌株26695和耐药株H390、e660、H57、H389的浮游菌具有抑制作用，且对26695和H390菌株的生物膜均具有抑制作用；盐酸小檗碱与阿莫西林对H. pylori的耐药菌株的联合抑菌作用为协同或相加作用；盐酸小檗碱可抑制上述26695和H390这2株H. pylori的浮游菌及生物膜脲酶活性；盐酸小檗碱可下调H. pylori浮游菌及生物膜的ureA、ureB基因的相对表达量，ureA、ureB基因的相对表达量与盐酸小檗碱共孵育的时间及浓度均呈递减的关系（P<0.05）。结论 盐酸小檗碱对H. pylori浮游菌及生物膜具有抑制作用，与阿莫西林联用对H. pylori起相加或协同作用；此外，其通过抑制脲酶活性并下调H. pylori的ureA、ureB基因表达，从而减少该菌浮游菌及生物膜的生长。
Abstract：Objective To investigate the effects of berberine hydrochloride on the expression of urease genes ureA and ureB in Helicobacter pylori(H. pylori) and its biofilm.Methods The standard strain 26695 of H. pylori, which was preserved in our laboratory, along with the drug-resistant strains H390, e660, H57, and H389 of H. pylori that were isolated from clinical sources were revived. The minimum inhibitory concentration(MIC) of berberine hydrochloride against the planktonic cells of the aforementioned five strains of H. pylori was determined using the broth dilution method. The combined inhibitory effect of berberine hydrochloride and amoxicillin against the aforementioned four strains of clinically resistant H. pylori was assessed using the checkerboard method. TheMIC of berberine hydrochloride against the biofilms of H. pylori strains 26695 and H390 was determined using the broth dilution method. Planktonic cells and biofilms of the aforementioned H. pylori strains 26695 and H390 were coincubated with berberine hydrochloride at concentrations of 0. 5 MIC, MIC, and 2 MIC for 2, 8, and 24 hours, respectively. Subsequently, urease activity was measured, and the expression levels of the ureA and ureB genes were detected using RT-qPCR.Results Berberine hydrochloride exhibited inhibitory effects against both planktonic cells of the antibiotic-sensitive strain 26695 and the antibiotic-resistant strains H390, e660, H57, and H389 of H. pylori. It also demonstrated inhibitory effects against the biofilms formed by strains 26695 and H390. Furthermore, the combined inhibitory effect of berberine hydrochloride and amoxicillin against antibiotic-resistant strains of H. pylori was either synergistic or additive. Berberine hydrochloride inhibited urease activity in both planktonic cells and biofilms of the H. pylori strains 26695 and H390. Furthermore, berberine hydrochloride downregulated the relative expression levels of the ureA and ureB genes in H. pylori and its biofilms. The relative expression levels of the ureA and ureB genes exhibited a dose-and time-dependent decrease with increasing concentrations of berberine hydrochloride and longer co-incubation times(P<0. 05).Conclusion Berberine hydrochloride inhibits both planktonic cells and biofilms of H. pylori. When combined with amoxicillin, it exerts additive or synergistic effects against H. pylori. Furthermore, by inhibiting urease activity and downregulating the expression of the ureA and ureB genes in H. pylori, berberine hydrochloride contributes to the reduction in growth of both planktonic cells and biofilms.
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基本信息:
DOI：10.19405/j.cnki.issn1000-1492.2026.08.004
中图分类号:R285
引用信息:
[1]罗婷婷,李梦沁,杨雪,等.盐酸小檗碱对幽门螺杆菌及其生物膜脲酶基因ureA、ureB表达的影响[J].安徽医科大学学报,2026,61(08):1355-1363.DOI:10.19405/j.cnki.issn1000-1492.2026.08.004.
基金信息:
贵州省科技计划项目（编号：ZK[2022]368、ZSYS[2025]021）; 贵州医科大学附属医院马歇尔联合实验室项目（编号：GYFYMF001、GYFYMF002）~~
2026-07-13
2026-07-13
2026-07-13