重组耻垢分枝杆菌联合抗痨药物对鼠结核病治疗作用的探讨  

Evaluation synergism of GLS/IL-12 recombinant M.s megmatis to anti-tuberculosis chemotherapy in murine model of M.tuberculosis infection

作　　者：杨晓燕[1,2] 杨春[3,2] 何永林[3,2] 徐蕾[3,2] 

机构地区：[1]重庆医科大学基础医学院免疫学教研室,重庆400016 [2]重庆医科大学基础医学院神经科学研究中心,重庆400016 [3]重庆医科大学基础医学院病原生物学教研室,重庆400016

出　　处：《重庆医科大学学报》2009年第5期516-520,共5页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(30400375)。

摘　　要：目的:探讨携带编码人天然颗粒溶素(Granulysin,GLS)和小鼠IL-12基因的重组耻垢分枝杆菌(M.smegmatis,MS)与抗痨药物对结核分枝杆菌感染的协同治疗效果。方法:将结核分枝杆菌H37Rv感染的Balb/c小鼠40只随机分成4组。感染4周后分别用生理盐水、重组MS、INH+PZA和重组MS联合INH+PZA治疗,于第一次治疗后3个月,处死小鼠,检测器官荷菌量、脾淋巴细胞IFN-γ和TNF-α的分泌水平、血清IL-12和IFN-γ和肺、脾组织中GLS表达,同时观察小鼠肺、脾组织病理改变情况。结果:重组MS联合INH+PZA治疗组肺、脾组织荷菌量(lgCFU/g)比重组MS和INH+PZA治疗组显著降低;重组MS和INH+PZA治疗组肺、脾组织荷菌量比生理盐水组显著降低。重组MS组经PPD刺激后IFN-γ和TNF-α分泌水平明显高于其它组。重组MS联合INH+PZA治疗组和重组MS组血清IL-12和IFN-γ水平明显高于生理盐水组。重组MS和INH+PZA组病变轻且局限,生理盐水组肺组织病理改变以渗出为主,病变广泛。重组MS联合INH+PZA治疗组肺组织病变最轻。结论:GLS/IL-12重组MS对小鼠结核病有一定免疫治疗作用;重组MS联合临床常用的抗痨药可增强抗结核病的疗效。Objective： To evaluate the synergism of recombinant M.smegmatis（ MS ）encoding human granulysin and murine IL-12 to anti-tuberculosis chemotherapy in murine M.tuberculosis M.tuberculosis infection. Methods： Balb/c mice were infected with normal saline, recombinant MS, INH （Isoniazid）＋PZA （pyrazinamide）, recombinant MS ＋ INH＋PZA respectively. The numbers of viable bacteria in the lung and the spleen were counted. The levels of IL12 and IFN- γ, in serum and IFN-γ and TNF-α released from spleen lymphocytes stimulated with PPD were detected with ELISA. Lungs and spleens were prepared for pathological analysis. Results： The recombinant MS group, INH＋PZA group, and recombinant MS ＋ IN H＋PZA group showed significantly reduced number of colony forming units （log CFU/g） compared with saline group.The increase of IL-12 and IFN- γ in serum were found in recombinant MS group. IFN- γ and TNF- α re- leased from spleen lymphocytes stimulated with PPD in recombinant MS group was higher than that in other groups （P〈0.01）. The pathological changes in lungs of the recombinant MS group, INH（Isoniazid ）＋PZA （pyrazinamide）group, and recombinant MS ＋INH＋PZA group were localized, while those in the control group were extensive. The significant pathological changes were not found in the spleens of all groups. Conclusion：The recombinant MS had immunotherapeutic effects, the recombinant MS had a synergism to anti-tuberculosis chemotherapy.

关 键 词：颗粒溶素 IL-12 重组MS 结核病 抗痨药 

分 类 号：R378.911[医药卫生—病原生物学]