Chen Meiyun, Wu Ziyao, Zou Yuanjing, Peng Chang, Hao Yajie, Zhu Zhixiang, Shi Xiaoyan, Su Bingmei, Ou Ling, Lai Yuqian, Jia Junwei, Xun Mingjin, Li Hui, Zhu Weixing, Feng Zhong, Yao Meicun
School of Pharmaceutical Sciences (Shenzhen), Sun Yat-sen University, Shenzhen, 518107, China.
School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, 510515, China.
J Ethnopharmacol. 2024 Oct 28;333:118396. doi: 10.1016/j.jep.2024.118396. Epub 2024 May 31.
Phellodendron chinense C.K.Schneid(P. chinense Schneid) is known in TCM as Huang Bo, is traditionally used to support gastrointestinal function and alleviate stomach-related ailments, including gastric ulcer bleeding and symptoms of gastroesophageal reflux disease. Helicobacter pylori (H. pylori) is classified by the WHO as a Group 1 carcinogen. However, the specific activity and mechanism of action of P. chinense Schneid against H. pylori infection remain unclear. It has been noted that Huangjiu processing may alter the bitter and cold properties of P. chinense Schneid, but its effect on antimicrobial activity requires further investigation. Additionally, it remains uncertain whether berberine is the sole antimicrobial active component of P. chinense Schneid.
This study aims to elucidate the anti-H. pylori infection activity of P. chinense Schneid, along with its mechanism of action and key antimicrobial active components.
Phytochemical analysis was carried out by UPLC-MS/MS. HPLC was employed to quantify the berberine content of the extracts. Antimicrobial activity was assessed using the micro broth dilution method. Morphology was observed using SEM. The impact on urease activity was analyzed through in vitro urease enzyme kinetics. RT-qPCR was employed to detect the expression of virulence genes, including adhesin, flagellum, urease, and cytotoxin-related genes. The adhesion effect was evaluated by immunofluorescence staining and agar culture.
P. chinense Schneid exhibited strong antimicrobial activity against both antibiotic-sensitive and resistant H. pylori strains, with MIC ranging from 40 to 160 μg/mL. Combination with amoxicillin, metronidazole, levofloxacin, and clarithromycin did not result in antagonistic effects. P. chinense Schneid induced alterations in bacterial morphology and structure, downregulated the expression of various virulence genes, and inhibited urease enzyme activity. In co-infection systems, P. chinense Schneid significantly attenuated H. pylori adhesion and urease relative content, thereby mitigating cellular damage caused by infection. Huangjiu processing enhanced the anti-H. pylori activity of P. chinense Schneid. Besides berberine, P. chinense Schneid contained seven other components with anti-H. pylori activity, with palmatine exhibiting the strongest activity, followed by jatrorrhizine.
This study sheds light on the potential therapeutic mechanisms of P. chinense Schneid against H. pylori infection, demonstrating its capacity to disrupt bacterial structure, inhibit urease activity, suppress virulence gene transcription, inhibit adhesion, and protect host cells. The anti-H. pylori activity of P. chinense Schneid was potentiated by Huangjiu processing, and additional components beyond berberine were identified as possessing strong anti-H. pylori activity. Notably, jatrorrhizine, a core component of P. chinense Schneid, exhibited significant anti-H. pylori activity, marking a groundbreaking discovery.
中药黄柏（Phellodendron chinense C.K.Schneid，即P. chinense Schneid）在传统中医中被称为黄柏，传统上用于支持胃肠功能并缓解与胃部相关的疾病，包括胃溃疡出血和胃食管反流病症状。幽门螺杆菌（Helicobacter pylori，简称H. pylori）被世界卫生组织列为1类致癌物。然而，黄柏对幽门螺杆菌感染的具体活性和作用机制尚不清楚。已经注意到，酒炙处理可能会改变黄柏的苦寒特性，但其对抗菌活性的影响需要进一步研究。此外，黄连素是否是黄柏唯一的抗菌活性成分仍不确定。
本研究旨在阐明黄柏对幽门螺杆菌感染的抗菌活性及其作用机制和关键抗菌活性成分。
采用超高效液相色谱-串联质谱法（UPLC-MS/MS）进行植物化学分析。采用高效液相色谱法（HPLC）对提取物中的黄连素含量进行定量。采用微量肉汤稀释法评估抗菌活性。使用扫描电子显微镜（SEM）观察形态。通过体外尿素酶酶动力学分析对尿素酶活性的影响。采用逆转录-定量聚合酶链反应（RT-qPCR）检测包括黏附素、鞭毛、尿素酶和细胞毒素相关基因在内的毒力基因的表达。通过免疫荧光染色和琼脂培养评估黏附效果。
黄柏对幽门螺杆菌敏感菌株和耐药菌株均表现出较强的抗菌活性，最低抑菌浓度（MIC）范围为40至160μg/mL。与阿莫西林、甲硝唑、左氧氟沙星和克拉霉素联合使用未产生拮抗作用。黄柏引起细菌形态和结构改变，下调多种毒力基因的表达，并抑制尿素酶活性。在共感染系统中，黄柏显著减弱幽门螺杆菌的黏附及尿素酶相对含量，从而减轻感染引起的细胞损伤。酒炙处理增强了黄柏的抗幽门螺杆菌活性。除黄连素外，黄柏还含有其他七种具有抗幽门螺杆菌活性的成分，其中巴马汀活性最强，其次是药根碱。
本研究揭示了黄柏对幽门螺杆菌感染的潜在治疗机制，证明其具有破坏细菌结构、抑制尿素酶活性、抑制毒力基因转录、抑制黏附以及保护宿主细胞的能力。酒炙处理增强了黄柏的抗幽门螺杆菌活性，并且鉴定出除黄连素外的其他成分具有较强的抗幽门螺杆菌活性。值得注意的是，黄柏的核心成分药根碱表现出显著的抗幽门螺杆菌活性，这是一项开创性的发现。
