| 纪文博 尚永强 臧师竹 张翠丽 姜涛.利用无传染性耻垢分枝杆菌建立小鼠结核病模型的探索[J].,2014,14(31):6065-6068 |
| 利用无传染性耻垢分枝杆菌建立小鼠结核病模型的探索 |
| Establishment of Mice Disease Models on Tuberculosis UsingNon-pathogenic Mycobacteria Smegmatis |
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| DOI： |
| 中文关键词: 耻垢分枝杆菌 C57BL/6 小鼠 结核病模型 |
| 英文关键词: Mycobacteria smegmatis C57BL/6 mice Tuberculosis models. |
| 基金项目:辽宁省" 挑战杯"大学生科技创新项目；高等学校博士学科点专项科研基金项目（20122105120014）；
辽宁省教育厅科学研究一般项目（L2012315） |
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| 中文摘要: |
| 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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