幽门螺杆菌表型与毒力的相关性研究  

Study on relationship between phenotype and virulence of Helicobacter pylori

作　　者：陈玉礼[1] 韦红玉[1] 唐华英[1] 赵丽娟[1] 曾怡[1] CHEN Yuli;WEI Hongyu;TANG Huaying;ZHAO Lijuan;ZENG Yi(Department of Microbiology and Immunology,Youjiang Medical University for Nationalities,Baise 533000,China)

机构地区：[1]右江民族医学院微生物与免疫学教研室,百色533000

出　　处：《右江医学》2018年第3期259-262,共4页Chinese Youjiang Medical Journal

基　　金：广西硕士研究生创新项目(201601002)

摘　　要：目的明确幽门螺杆菌表型与毒力的相关性,掌握幽门螺杆菌的生物学特征,为临床快速了解幽门螺杆菌的毒力并通过减毒方法防治幽门螺杆菌提供实验依据。方法配制含有合适浓度的甲硝唑培养液,用倍比稀释法使培养液中甲硝唑的浓度为0μg/ml、2μg/ml、4μg/ml、8μg/ml、16μg/ml、32μg/ml,分别培养幽门螺杆菌,诱导幽门螺杆菌的形态发生球形变,然后用革兰染色镜下观察培养12 h、24 h、48 h时在不同抗生素浓度下幽门螺杆菌形态学的变化,并用Real-time PCR法检测不同形态学下其细胞毒素相关基因A(Cag A)和空泡毒素基因(Vac A)毒力基因的表达。结果培养12 h时幽门螺杆菌的形态未发生变化;当培养24 h时在16μg/ml、32μg/ml浓度抗生素下,一部分幽门螺杆菌由螺旋状变成了短杆状;培养48 h时在16μg/ml、32μg/ml浓度抗生素下,幽门螺杆菌几乎全部变成了圆球状,其他抗生素浓度下的幽门螺杆菌没有形态学变化,且球形变时幽门螺杆菌的Cag A和Vac A表达下降。结论幽门螺杆菌表型与毒力有密切的关系,抗生素能诱导幽门螺杆菌发生形态学变化且球形变的幽门螺杆菌毒力下降。Objective To clarify relationship between the phenotype and virulence of Helicobacter pylori and to grasp the biological characteristics of Helicobacter pylori,so as to provide experimental evidence for the clinical understanding of the virulence of Helicobacter pylori and the prevention and treatment of it by detoxification. Methods Metronidazole medium containing suitable concentration was prepared.The concentration of metronidazole in the culture medium was 0 μg/ml,2 μg/ml,4 μg/ml,8 μg/ml,16 μg/ml,32 μg/ml by double dilution method,and they were respectively used to culture Helicobacter pylori.The morphology of Helicobacter pylori was induced to be spherical. And then,the morphological changes of Helicobacter pylori at different concentrations of antibiotics were observed under Gram staining microscope at 12 h,24 h and 48 h.In addition,the expression of cytotoxin associated gene（ Cag A） and vacuolating cytotoxin gene A（ Vac A） under different morphology were detected by real time PCR.Results The morphology of Helicobacter pylori did not change when culturing for12 h,while some Helicobacter pylori changed from spiral to short rod under concentrations of 16 g/ml and 32 g/ml when culturing for 24 h.When culturing for 48 h,almost all of the Helicobacter pylori became globular at the concentration of 16 μg/ml and 32 μg/ml,but no morphological changes were found in other concentrations of antibiotics.Furthermore,the expression of Cag A and Vac A genes in Helicobacter pylori decreased during globular change.Conclusion The phenotype of Helicobacter pylori is closely related to its virulence.Antibiotics can induce morphological changes of Helicobacter pylori and decrease the virulence of the globular Helicobacter pylori.

关 键 词：幽门螺杆菌 抗生素 表型 毒力变化 

分 类 号：R378[医药卫生—病原生物学]