新生隐球菌对鼠小胶质细胞细胞骨架蛋白质组的影响

Different expression of microglia N9 cytoskeletal proteome in the interaction between Cryptococcus neoformans and murine microglia

目的:探讨新生隐球菌对鼠小胶质细胞(N9)蛋白质组的影响。方法:分别提取正常N9细胞以及与新生隐球菌(ATCC32609)作用后的N9细胞的蛋白质样品,采用双向电泳技术分离,对部分差异的蛋白质点进行基质辅助激光解吸电离飞行时间质谱分析,测定其肽质量指纹图谱,在网上蛋白质数据库检索鉴定差异蛋白。结果:获得较好的双向电泳图像,图像分析显示:在与新生隐球菌作用前、后检测到N9细胞的蛋白质点分别是(701±29)个、(980±20)个,对表达差异的18个蛋白质点进行肽质量指纹图谱分析,经数据库检索后,鉴定出3个涉及小胶质细胞细胞骨架的蛋白质:anillin、pericentrin 2、核苷二磷酸激酶A[nucleoside diphosphate kinase A(NDP kinase A,NDPKA)]。结论:新生隐球菌对小胶质细胞的细胞骨架蛋白质造成影响,这有助于在分子水平理解隐球菌性脑膜炎的发病机制。

Objective： To investigate the effects of Cryptococcus neoformans on murine microglia N9 proteome. Methods： Protein samples were extracted from normal N9 cells and N9 cells co-cultured with Cryptococcus neoformans. The proteins were separated by immobilized pH gradient two-dimensional（2-D） gel electrophoresis. The 2-D electrophoretogram was analyzed using ImageMaster software. The differential spots were identified by peptide mass fingerprinting based on matrix-assisted laser desorption ionization time of flight mass spectrometry（MALD-TOF-MS） and database searching was performed on the internet. Results： The 2-D image was stable and showed （701±29） spots in normal N9 cell sample and （980±20） spots in co-cultured N9 cells,respectively. Three of 18 differential spots were identified： anillin, pericentrin 2 and nucleoside diphosphate kinase A （NDP kinase A, NDPKA）. The three proteins correlated with cellular skeletal protein of N9 cell. Conclusion： Cryptococcus neoformans may have effects on the cytoskeleton proteins of murine microglia N9 cell which might be helpful to understand the molecular mechanisms of cryptococcal meningitis.