| 郭旭光1,2 夏勇1 马越云2 郝晓柯2.肺上皮细胞自噬体在结核分枝杆菌感染中保护作用的分子机制[J].,2011,11(21):4019-4023 |
| 肺上皮细胞自噬体在结核分枝杆菌感染中保护作用的分子机制 |
| The Effect of Autophagy on Mycobacterium Tuberculosis Infections ofLung Epithelial Cells and its Mechanism |
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| DOI： |
| 中文关键词: 自噬 自噬体 结核分枝杆菌 肺泡上皮细胞 |
| 英文关键词: Autophagy Autophagosome Mycobacterium Tuberculosis Lung Epithelial Cell |
| 基金项目:国家自然科学基金面上项目（30872358）广州医学院青年基金项目（2010A026） |
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| 中文摘要: |
| 目的：构建Atg5- 真核表达载体并瞬时转染肺上皮细胞细胞株，探讨自噬在结核分枝杆菌感染上皮细胞中保护作用的分子
机制。方法：设计针对Atg5的RNAi序列，化学合成后经过变性，退火连接到pSilencerTM 3.1-H1 hygro真核表达载体，经测序验证
其正确性。脂质体法瞬时转染真核细胞A549，免疫印迹法检测瞬时转染的效果。用结核分枝杆菌分别感染正常和自噬表达低下的
A549细胞，通过检测LDH 来观察细胞的坏死情况。结果：成功的构建了pSilencerTM 3.1-H1 hygro 真核表达载体并建瞬时转染了
A549细胞株，成功抑制了细胞的自噬功能。Atg5-细胞对结核杆菌的抵抗能力下降。结论：在自噬表达低下的细胞中，细胞对结核
分枝杆菌的抵抗能力有明显下降。在结核分枝杆菌感染上皮细胞的过程中，自噬是一种保护机制。 |
| 英文摘要: |
| Objective: To construct the eukaryotic plasmid of Atg5 and transfect A549 cells, and to investigate the role of autophagy
on Mycobacterium Tuberculosis infections of lung epithelial cells and its mechanism. Methods: The RNAi primer of Atg5 was
designed, and the Oligos of 64 base pairs for hairpin RNA targeting Atg5 were synthesized. Eukaryotic victor pSilencerTM 3.1-H1 hygro
was connected and DNA sequencing. The recombinant vector was transfected into A549 cells by lipofectamineTM 2000. Transient transfected
A549 cell line was established and identified by Western blot. Results: The eukaryotic expression vector pSilencerTM 3.1-H1 hygro
was constructed, and transient transfected A549 cell line was established. The protein was inhibited successfully. A549 cells and transient
transfected A549 cell were infected with Mycobacterium Tuberculosis, and the LDH were detected. Conclusions: The capacity of A549
to resistance of Mycobacterium Tuberculosis was declined significantly in the cells with low autophagy. Autophagy is a defense mechanism
in infected lung epithelial cells. |
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