巨噬细胞过氧化物酶体与鼠伤寒沙门菌相互作用的初步分析

发布时间：2018-04-19 00:30

本文选题：巨噬细胞 + 过氧化物酶体类　；参考：《解放军医学杂志》2008年04期

【摘要】：目的观察巨噬细胞过氧化物酶体在鼠伤寒沙门菌感染中的作用。方法将感染及未感染鼠伤寒沙门菌的鼠巨噬细胞RAW264.7以氮气压迫法裂解,逐级离心,用制备的TassC多抗磁珠纯化得到粗提液,用Western blot法鉴定TassC多抗磁珠与细胞结合物的性质;用pDs Red2-Perxi质粒转染RAW264.7细胞以标记细胞中的过氧化物酶体,用三维荧光显微镜观察感染细胞中过氧化物酶体(红色)与表达绿色荧光蛋白的鼠伤寒沙门菌突变株的相互作用。结果Western blot分析显示TassC多抗磁珠可以结合细胞内的TassC,并与细胞内的过氧化物酶体结合,也可以与感染细胞中的iNOS结合,而未感染样本中未检测到iNOS。磁珠洗脱物中未检测到溶酶体标志物LAMP1,也未检测到受染菌标志物;免疫荧光显示感染的巨噬细胞中过氧化物酶体聚集在鼠伤寒沙门菌突变株SCV周围,甚至发生重叠,随感染时间延长(1h或24h),过氧化物酶体数量增加(1.2倍或1.3倍),而胞内菌数量下降。结论TassC可能定位于过氧化物酶体而不是溶酶体;感染鼠伤寒沙门菌的巨噬细胞中iNOS可能与过氧化物酶体结合而参与杀灭微生物的作用。
[Abstract]:Objective to observe the role of macrophage peroxisome in the infection of Salmonella typhimurium.Methods the RAW264.7 of murine macrophages infected and uninfected with Salmonella typhimurium was decomposed by nitrogen pressure method and centrifuged step by step. The crude extract was obtained by purification of prepared TassC multi-diamagnetic beads. The properties of TassC multi-diamagnetic beads and cell conjugates were identified by Western blot method.RAW264.7 cells were transfected with pDs Red2-Perxi plasmid to label peroxisome. The interaction of peroxisome (red) in infected cells with mutant strain of Salmonella typhimurium expressing green fluorescent protein was observed by three dimensional fluorescence microscope.Results Western blot analysis showed that TassC multi-diamagnetic beads could bind to TassCin, peroxisome and iNOS in infected cells, but no iNOS was detected in uninfected samples.No lysosomal marker LAMP1 or bacterial marker was detected in magnetic bead elution, and the peroxisome in infected macrophages gathered around and even overlapped around the mutant strain of Salmonella typhimurium by immunofluorescence.The number of peroxisome increased 1.2 times or 1.3 times, while the number of intracellular bacteria decreased with the increase of infection time for 1 h or 24 h.Conclusion TassC may be located in peroxisome rather than lysosome, and iNOS in macrophages infected with Salmonella typhimurium may play a role in killing microorganisms by binding to peroxisome.
【作者单位】：
【基金】：国家自然科学基金资助课题(30170891) 全军“十五”科研规划面上项目资助课题(01MA144)
【分类号】：R378

【相似文献】