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经验性中药配方诃子清油的抗炎活性及潜在机制

anti- activity and the underlining mechanism of an empirical herbal formula - Hezi Qingyou.

作者信息

Feng Zhong, Li Hui, Hao Yajie, Peng Chang, Ou Ling, Jia Junwei, Xun Mingjin, Zou Yuanjing, Chen Meiyun, Zhang Guimin, Yao Meicun

机构信息

School of Pharmaceutical Sciences, Sun Yat-sen University, Shenzhen, China.

State Key Laboratory of Integration and Innovation of Classic Formula and Modern Chinese Medicine, Lunan Pharmaceutical Group Co., Ltd., Linyi, China.

出版信息

Front Microbiol. 2024 Feb 19;15:1355460. doi: 10.3389/fmicb.2024.1355460. eCollection 2024.

Abstract

BACKGROUND

() is thought to primarily colonize the human stomach and lead to various gastrointestinal disorders, such as gastritis and gastric cancer. Currently, main eradication treatment is triple or quadruple therapy centered on antibiotics. Due to antibiotic resistance, the eradication rate of is decreasing gradually. Therefore, searching for anti- drugs from herbal sources has become a strategy for the treatment. Our team proposed a Hezi Qingyou Formula (HZQYF), composed of Chebulae Fructus, Ficus hirta Vahl and Cloves, and studied its anti- activity and mechanism.

METHODS

Chemical components of HZQYF were studied using UHPLC-MS/MS and HPLC. Broth microdilution method and agar dilution method were used to evaluate HZQYF's antibacterial activity. The effects of HZQYF on expression of adhesion genes (, , ), urease genes (, ), and flagellar genes () were explored using Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR) technology. Effects on morphology and permeability of the extracellular membrane were studied using scanning electron microscopy (SEM) and N-phenylnaphthalen-1-amine (NPN) uptake. Effect on urease activity was studied using a urease kinetics analysis . Immunofluorescence staining method was used to examine the effect on adhesion. Western blot was used to examine the effect on cagA protein.

RESULTS

Minimum inhibitory concentration (MIC) values of the formula against clinical strains and standard strains were 80-160 μg/mL, and minimum bactericidal concentration (MBC) values were 160-320 μg/mL. The formula could down-regulate the expression of adhesion genes (A, , ), urease genes (, ) and flagellar genes (, ), change the morphology of , increase its extracellular membrane permeability, and decrease its urease activity.

CONCLUSION

Present studies confirmed that HZQYF had promising anti- activities and demonstrated its possible mechanism of action by down-regulating the bacterial adhesion, urease, and flagellar gene expression, which provided scientific bases for further clinical investigations.

摘要

背景

(某种细菌名称未给出)被认为主要定植于人的胃部,并导致各种胃肠道疾病,如胃炎和胃癌。目前,主要的根除治疗是以抗生素为中心的三联或四联疗法。由于抗生素耐药性,(该细菌名称)的根除率正在逐渐下降。因此,从草药来源寻找抗(该细菌名称)药物已成为一种治疗策略。我们的团队提出了一种由诃子、五指毛桃和丁香组成的诃子清优方(HZQYF),并研究了其抗(该细菌名称)活性及作用机制。

方法

采用超高效液相色谱-串联质谱法(UHPLC-MS/MS)和高效液相色谱法(HPLC)研究HZQYF的化学成分。采用肉汤微量稀释法和琼脂稀释法评估HZQYF的抗菌活性。运用逆转录定量聚合酶链反应(RT-qPCR)技术探究HZQYF对黏附基因(具体基因名称未给出)、脲酶基因(具体基因名称未给出)和鞭毛基因(具体基因名称未给出)表达的影响。使用扫描电子显微镜(SEM)和N-苯基萘-1-胺(NPN)摄取法研究对细胞外膜形态和通透性的影响。采用脲酶动力学分析法研究对脲酶活性的影响。使用免疫荧光染色法检测对黏附的影响。采用蛋白质免疫印迹法检测对cagA蛋白的影响。

结果

该方剂对(该细菌名称)临床菌株和标准菌株的最低抑菌浓度(MIC)值为80 - 160μg/mL,最低杀菌浓度(MBC)值为160 - 320μg/mL。该方剂可下调黏附基因(具体基因名称未给出)、脲酶基因(具体基因名称未给出)和鞭毛基因(具体基因名称未给出)的表达,改变(该细菌名称)的形态,增加其细胞外膜通透性,并降低其脲酶活性。

结论

目前的研究证实HZQYF具有良好的抗(该细菌名称)活性,并通过下调细菌黏附、脲酶和鞭毛基因表达证明了其可能的作用机制,为进一步的临床研究提供了科学依据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32bd/10910045/fee8a80bcd7b/fmicb-15-1355460-g001.jpg

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