黄芩提取物对幽门螺杆菌形态及生理生化特性的抑制作用研究  

Inhibitory effect of Scutellaria baicalensis Georgi on morphology,physiological and biochemical characteristics of Helicobacter pylori

作　　者：张建辉 沈姝霖 李雯慧 王志强[1] 季晓飞[1] 赵慧琳[1] 张艳丽[1] ZHANG Jianhui;SHEN Shulin;LI Wenhui;WANG Zhiqiang;JI Xiaofei;ZHAO Huilin;ZHANG Yanli(Binzhou Medical University,Yantai,Shandong 264003,China)

机构地区：[1]滨州医学院,山东烟台264003

出　　处：《中国病原生物学杂志》2024年第10期1172-1176,共5页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.81702054);山东省自然科学基金(No.ZR2020MH297,ZR2023MH101);山东省高校青年创新科技支持计划(No.2020KJK006)。

摘　　要：目的 明确黄芩提取物(TESB)对幽门螺杆菌(Hp)的抑制作用,为其临床上应用于Hp感染的治疗提供实验依据。方法 通过在琼脂平板表面混合涂布TESB油溶液和Hp并对其进行连续传代,建立TESB处理Hp模型,利用光学显微镜、扫描电镜对比观察TESB对Hp菌体形态结构的影响;通过测定生长曲线,分析TESB对Hp生长速率的影响;通过RT-qPCR检测Hp重要毒力因子尿素酶、CagA、VacA的表达水平,通过尿素酶实验检测Hp的尿素酶活性,分析TESB对Hp毒力因子的影响;构建表达绿色荧光蛋白(GFP)的Hp突变菌株,并以之感染胃上皮细胞GES-1,通过检测荧光强度,比较TESB处理前后Hp对细胞的吸附能力。结果 通过固体平板混合涂布并连续传代7代后成功建立Hp的TESB处理模型(Hp~(TESB)),光学显微镜和扫描电镜观察显示,加药组菌体出现明显球形变异,生长明显滞后,且细菌最高生长浓度远低于未处理组;RT-qPCR结果显示,主要毒力因子表达量均下降;细胞实验显示,药物处理组的细菌对细胞吸附能力减弱,但将药物去除后,细菌的形态可逐渐恢复为螺旋状。结论 TESB能够将Hp从正常的螺旋形态转变为休眠的球状形态,抑制Hp的生长,下调其毒力因子的表达水平,并降低其对细胞的黏附能力。但这一抑制作用是可逆的,临床应用时需考虑与其他药物联合使用。Objective To clarify the effect of Scutellaria baicalensis Georgi(TESB)on reducing Hp physiological activities and virulence,and provide experimental foundation for the clinical application of TESB in treatment of Hp infection.Methods The model of Hp treated with TESB was established by coating agar plate with mixed TESB and Hp for serial passaging.The effects of TESB on the morphology and structure of Hp cells were compared by optical microscope and scanning electron microscope.The effects of TESB on the growth of Hp were compared by measuring the growth curve.The gene expression levels of Hp virulence factors,including urease,CagA and VacA,were detected by urease experiment and RT-qPCR to evaluate the influence of TESB on Hp virulence factors.Finally,an Hp mutant strain expressing green fluorescent protein(GFP)was constructed to infect gastric epithelial cells(GES-1).The adsorption capacity of Hp to cells before and after TESB treatment was compared by detecting fluorescence intensity.Results TESB treated Hp model(HpTESh)was successfully established after 7 generations passaging of mixed cultivation of Hp and TESB on solid plate.Observation by light microscope and Scanning Electron Microscope revealed that TESB treated bacteria showed an obvious coccoid alteration and growth lag,and the maximum bacterial concentration was much lower than that in untreated group.RT-qPCR indicated that the expression levels of major virulence factors and adsorption ability of TESB treated bacteria were decreased.However,after TESB is removed,the shape of the bacteria can gradually return to spiral.Conclusion TESB can transform Hp from the normal spiral shape to the dormant coccoid form,inhibit the growth of Hp,down-regulate the expression level of its virulence factors,and reduce its adhesion to cells.But this inhibitory effect is reversible,other drugs should be considered to added in combination with TESB in clinical application.

关 键 词：黄芩提取物 幽门螺杆菌 致病因子 抑菌 

分 类 号：R285[医药卫生—中药学]