海分枝杆菌感染斑马鱼成鱼模型的构建  

Establishment of adult zebrafish model of M.marinum infection

作　　者：杨国威 彭世泽 张永欣[2] 赵建元[2] 岑山[2] YANG Guo-wei;PENG Shi-ze;ZHANG Yong-xin;ZHAO Jian-yuan;CEN Shan(Beijing Tropical Medicine Research Institute,Beijing Friendship Hospital Affiliated to Capital Medical University,Beijing 100050,China;Department of Immunology,Institute of Medicinal Biotechnology,Chinese Academy of Medical Science & Peking Union Medical College,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京友谊医院北京热带医学研究所,北京100050 [2]中国医学科学院北京协和医学院医药生物技术研究所免疫生物学室,北京100050

出　　处：《中国医药生物技术》2019年第3期224-228,共5页Chinese Medicinal Biotechnology

基　　金：中国医学科学院医学与健康创新工程(2017-I2M-2-002 YXZ)

摘　　要：目的构建海分枝杆菌感染斑马鱼模型,对先导化合物的有效性及其代谢衍生物进行检测,以有效降低药物体内筛选的成本。方法采用海分枝杆菌感染斑马鱼成鱼,测定斑马鱼的存活率,用HE以及Ziehl-Neelsen染色的方式,观察斑马鱼体内肉芽肿形成情况,并对形成的肉芽肿数进行计数,用于评价抗结核药物利福平对斑马鱼成鱼结核病的治疗效果,结合细菌菌落定量PCR计算利福平给药的IC50值。结果成功构建了海分枝杆菌感染斑马鱼的抗结核药物筛选模型。利福平可以减少海分枝杆菌感染斑马鱼体内的肉芽肿数目和细菌数,对照组肉芽肿数目平均有28个,200μmol/L利福平给药组有5.6个,而利福平高剂量组未发现肉芽肿,两个治疗组与对照组相比均有显著性差异(P<0.05)。利福平成鱼给药的IC50值为52.5μmol/L。结论海分枝杆菌-斑马鱼感染模型结合细菌菌数定量PCR分析方法是一种简单、高效的抗结核药物体内筛选模型,可用于抗结核药物的筛选与评价。Objective We aim to establish a zebrafish model of M.marinum infection,which can detect the effectiveness of leading compounds and their metabolic derivatives in a complete in vivo physiological environment,and thus greatly reduce costs of drug screening.Methods Adult zebrafish were infected with M.marinum,then the survival rate,numbers of bacteria and morphology of granuloma of infected zebrafish were determined.Moreover,the anti-tuberculosis drug rifampicin was tested for treatment of tuberculosis infected zebrafish and IC50 value of rifampicin was calculated by quantitative PCR.Results The potential anti-tuberculosis drugs based on zebrafish model of M.marinum infection was successfully constructed.Rifampicin can effectively reduce the number of granuloma and bacteria in zebrafish infected by M.marinum,which is significantly different from the control group(P<0.05),and the IC50 value for curing adult fish is 52.5μmol/L.Conclusion Zebrafish model of M.marinum infection combined with bacterial quantitative PCR analysis is a simple and efficient in vivo screening and evaluation system for the discovery of potential anti-tuberculosis drugs.

关 键 词：结核 海分枝杆菌 斑马鱼 抗结核药 

分 类 号：R52[医药卫生—内科学] R-332[医药卫生—临床医学]