幽门螺杆菌（HP）是胃中最常见的致病微生物，可诱发胃粘膜炎症反应，引起慢性胃炎甚至胃癌。 HP感染影响全球超过44亿人，全球感染率高达50%。 HP的多重耐药性给根除带来了严峻的挑战。事实证明，与铋剂四联疗法相比，清热化湿汤联合三联疗法的根除率相当，但不良反应发生率较低；此外，QHD在体外可直接抑制和杀灭HP。探讨QHD对临床多重耐药和强生物膜形成HP的作用及机制。本研究在HP胃镜检查时活检后体外分离12株HP菌株- 感染的患者。在体外，分别通过E-test方法和结晶紫染色确定临床HP菌株的最低抑菌浓度（MIC）值和生物膜定量。选择HP生物膜形成能力最强的菌株，评估QHD对生物膜形成能力最强的菌株的抑制和杀菌效果。该评估是使用琼脂稀释、电子测试、杀伤动力学和透射电子显微镜 (TEM) 进行的。该研究还探讨了 QHD 对这些具有强烈生物膜形成的 HP 菌株抗生素耐药性的影响。采用结晶紫法、扫描电子显微镜、激光共焦扫描显微镜和(p)ppGpp色谱鉴定方法评价QHD对强生物膜形成HP菌株生物膜的影响。通过定量聚合酶链反应评估 QHD 对生物膜和外排泵相关基因表达的影响。采用UHPLC-MS/MS非靶向代谢组学技术鉴定NC组和QHD组之间不同的潜在代谢途径和生物标志物。HP在体外可以形成不同程度的生物膜，形成的强度与耐药性相关的应变。 QHD对HP具有较强的抑菌和杀菌作用，MIC为32-64 mg/mL。 QHD可以抑制强生物膜形成HP菌株的生物膜形成，破坏生物膜结构，降低(p)ppGpp的积累，减少生物膜相关基因的表达，包括LuxS、Spot、glup (HP1174)、NapA和CagE，并减少外排泵相关基因的表达，例如 HP0605、HP0971、HP1327 和 HP1489。根据代谢组学分析，QHD可诱导HP氧化应激，增强代谢，并可能通过上调腺苷单磷酸（AMP）抑制相关信号通路，从而影响HP生长、代谢和蛋白质合成。QHD对HP发挥抑菌和杀菌作用，并且通过抑制HP生物膜形成、破坏其生物膜结构、抑制生物膜相关基因和外排泵相关基因的表达、增强HP代谢、激活HP中的AMP来降低HP耐药性。©作者2024。出版者百事登出版集团有限公司版权所有。

Helicobacter pylori (HP), the most common pathogenic microorganism in the stomach, can induce inflammatory reactions in the gastric mucosa, causing chronic gastritis and even gastric cancer. HP infection affects over 4.4 billion people globally, with a worldwide infection rate of up to 50%. The multidrug resistance of HP poses a serious challenge to eradication. It has been de-monstrated that compared to bismuth quadruple therapy, Qingre Huashi decoction (QHD) combined with triple therapy exhibits comparable eradication rates but with a lower incidence of adverse reactions; in addition, QHD can directly inhibit and kill HP in vitro.To explore the effect and mechanism of QHD on clinically multidrug-resistant and strong biofilm-forming HP.In this study, 12 HP strains were isolated in vitro after biopsy during gastroscopy of HP-infected patients. In vitro, the minimum inhibitory concentration (MIC) values for clinical HP strains and biofilm quantification were determined through the E-test method and crystal violet staining, respectively. The most robust biofilm-forming strain of HP was selected, and QHD was evaluated for its inhibitory and bactericidal effects on the strain with strong biofilm formation. This assessment was performed using agar dilution, E-test, killing dynamics, and transmission electron microscopy (TEM). The study also explored the impact of QHD on antibiotic resistance in these HP strains with strong biofilm formation. Crystalline violet method, scanning electron microscopy, laser confocal scanning microscopy, and (p)ppGpp chromatographic identification were employed to evaluate the effect of QHD on biofilm in strong biofilm-forming HP strains. The effect of QHD on biofilm and efflux pump-related gene expression was evaluated by quantitative polymerase chain reaction. Non-targeted metabolomics with UHPLC-MS/MS was used to identify potential metabolic pathways and biomarkers which were different between the NC and QHD groups.HP could form biofilms of different degrees in vitro, and the intensity of formation was associated with the drug resistance of the strain. QHD had strong bacteriostatic and bactericidal effects on HP, with MICs of 32-64 mg/mL. QHD could inhibit the biofilm formation of the strong biofilm-forming HP strains, disrupt the biofilm structure, lower the accumulation of (p)ppGpp, decrease the expression of biofilm-related genes including LuxS, Spot, glup (HP1174), NapA, and CagE, and reduce the expression of efflux pump-related genes such as HP0605, HP0971, HP1327, and HP1489. Based on metabolomic analysis, QHD induced oxidative stress in HP, enhanced metabolism, and potentially inhibited relevant signaling pathways by upregulating adenosine monophosphate (AMP), thereby affecting HP growth, metabolism, and protein synthesis.QHD exerts bacteriostatic and bactericidal effects on HP, and reduces HP drug resistance by inhibiting HP biofilm formation, destroying its biofilm structure, inhibiting the expression of biofilm-related genes and efflux pump-related genes, enhancing HP metabolism, and activating AMP in HP.©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.