颗粒溶素对结核分枝杆菌感染小鼠的保护效应  

Protective Effect of Granulysin on Mice Infected with Mycobacterium Tuberculosis

作　　者：刘斌波[1] 刘君炎[2] 刘胜武[2] 屈雪菊[2] 

机构地区：[1]武汉大学医学院微生物学教研室,湖北武汉430071 [2]武汉大学医学院免疫学教研室,湖北武汉430071

出　　处：《武汉大学学报（医学版）》2011年第3期292-295,F0002,共5页Medical Journal of Wuhan University

基　　金：湖北省科技厅重大课题资助项目(编号:301121028);湖北省自然科学基金资助项目(编号:301131738)

摘　　要：目的:构建颗粒溶素(GNLY)真核表达质粒,并将重组质粒肌肉注射结核分枝杆菌感染小鼠观察其保护效应。方法:将人颗粒溶素cDNA插入pcDNA3.1(-)构建真核表达质粒。18只感染结核分枝杆菌H37Rv株的BALB/c小鼠随机分为生理盐水组、pcDNA3.1(-)质粒组和GNLY质粒组。感染第1天和第6天各组小鼠分别从后腿股四头肌注射生理盐水、pcDNA3.1(-)质粒、GNLY质粒,免疫2次。感染4周后,处死各组小鼠,检测器官活菌数[lgCFU/(g.ml)],脾重指数(WI),并观察肺、脾组织病理情况及抗酸染色情况。结果:GNLY质粒组肺组织活菌数为6.688±0.003,显著低于生理盐水组(7.821±0.090,P<0.01)和pcDNA3.1(-)质粒组(7.848±0.017,P<0.01);GNLY质粒组脾组织活菌数为7.140±0.034,显著低于两对照组(7.805±0.080,7.858±0.043,P<0.01)。GNLY质粒组脾重指数(2.575±0.649)也显著低于生理盐水组(4.580±0.967,P<0.01)和pcDNA3.1(-)质粒组(3.885±0.228,P<0.01)。GNLY质粒组肺脏病理学损害明显轻于两对照组,组织抗酸染色细菌荷载数量明显较两对照组低。结论:GNLY对结核分枝杆菌感染小鼠具有一定的保护作用。Objective： To construct eukaryotic expression vector for granulysin and to study its protective effect on mice infected with Mycobacterium tuberculosis.Methods： A full length of cDNA of granulysin was inserted into pcDNA3.1（-） vector to construct eukaryotic expression plasmid for granulysin.Eighteen BALB/c mice infected with Mycobacterium tuberculosis were randomized into three groups： NS group,pcDNA3.1（-） group and granulysin recombinant plasmids group.After one day and six days respectively of challenging with Mycobacterium tuberculosis from lateral tail,mice were intramuscular immunized with normal saline,pcDNA3.1（-） and granulysin recombinant plasmids twice respectively.Four weeks after infection,mice were sacrificed,the lungs and spleens were removed and the numbers of viable bacteria（lgCFU/） in organs and histopathologic changes were determined.Results： In granulysin recombinant plasmids group,the numbers of viable bacteria in lung were 6.688±0.003 and 7.140±0.034 in spleen,and the weight index of spleen was 2.575±0.649.There were significant difference between control groups and granulysin recombinant plasmids group.The lesion and the bacterial loading of lung tissues in granulysin recombinant plasmids group were significantly slighter than those of the two control groups.Conclusion： Granulysin has protective effect against Mycobacterium tuberculosis infection.

关 键 词：结核分枝杆菌 颗粒溶素 真核表达质粒 治疗作用 

分 类 号：R378.91[医药卫生—病原生物学]