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目的：利用耻垢分枝杆菌（M.smegmatis mc2155）建立C57BL/6 小鼠结核病模型。方法：每天以高剂量（5 × 107 CFU）耻垢分枝杆菌给C57BL/6 小鼠腹腔注射，连续感染4 周，检测耻垢分枝杆菌对小鼠的致病性。分别于2 周和4 周处死小鼠，无菌条件下解剖小鼠取肺、脾脏组织匀浆，进行组织内细菌活力检测；通过嗜酸性染色进行分枝杆菌的鉴定；同时进行病理切片的制备，观察肺和脾脏组织的病理变化；最后进行菌体DNA的提取和基因检测，根据上述指标确定小鼠结核病模型的建立是否成功。结果：腹腔感染小鼠2 周后, 模型组小鼠只有脾脏组织匀浆液出现抗酸染色阳性菌落，肺部组织未见阳性菌落。腹腔感染小鼠4 周后, 模型组小鼠肺、脾脏组织匀浆液中均可见大量抗酸染色阳性的菌落；组织病理学观察结果显示：小鼠肺组织主要表现为以中性粒细胞为主的炎性病变；基因检测结果表明：模型组小鼠肺组织匀浆液中可检测到耻垢分枝杆菌特异性3- 磷酸甘油醛脱氢酶（gap）基因，而脾脏组织未扩增出耻垢分枝杆菌特异性基因。结论：通过腹腔注射无致病性耻垢分枝杆菌方法，成功建立C57BL/6 小鼠结核病发生模型，为结核分枝杆菌与宿主相互作用研究提供安全的疾病模型。

英文摘要:

Objective:To establish tuberculosis mice models by using no pathogenic mycobacteria smegmatis mc2155.Methods:High dose mycobacteria smegmatis mc2155 (5× 107 CFU) were injected in mice abdominal of C57BL / 6 cavity injection once per day for 4 successive weeks, so that to observe the pathogenic of mycobacteria smegmatis in mice. Inflammatory changes of lung tissue sections were observed by hematoxylin and eosin stain (HE stainning); Specibility acid-fast staining method was used to identify infection capability and vitality of mycobacteria smegmatis in C57BL/6 mice lung and spleen tissue after infection 2 week and 4 week. Finally, bacteria DNA in lung and spleen tissue cuture was extracted and mycobacterial smegmatis specific glyceraldehyde 3 - phosphoric acid dehydrogenase gene (gap) were amplied by using PCR method. The feasibility of using non-infectious mycobacteria smegmatis to establish tuberculosis mice models were explored according to the above three aspects detection.Results:At the time of infection 2 week, it was observed rod shape mycobacteria smegmatis by using specibility oxychromatic staining in C57BL/6 mice spleen tissue culture, instead of the lung tissue. After 4 week, pathological changes for most of the alveolar cavity filled with neutrophils inflammatory lesions. Rod shape bacteria were observed by using specibility acid-fast staining in C57BL/6 mice lung tissue and spleen tissue culture. The specific glyceraldehyde 3-phosphoric acid dehydrogenase gene (gap) of mycobacteria smegmatis was amplied in the lung homogenate of model mice, instead spleen tissue and the control group was not.Conclusion:C57BL / 6 mice tuberculosis model was established successfully by intraperitoneal injection method of non-pathogenic mycobacteria smegmatis. This method provides security disease models for interaction studies about mycobacteriumtuberculosis and host cell.

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