猪链球菌2型溶菌酶释放蛋白诱导血管内皮细胞融合-中国农业科学.PDF

中国农业科学 2005,38(4):821-825 Scientia Agricultura Sinica 猪链球菌2型溶菌酶释放蛋白诱导血管内皮细胞融合 1，2 1 曾巧英 ， 陆承平 1 2 ( 南京农业大学农业部动物疫病诊断与免疫重点开放实验室，南京 210095 ； 甘肃农业大学动物医学院，兰州 730070 ） 摘要：猪链球菌2型（SS2）主要引起人和猪的脑膜炎，但其毒力因子溶菌酶释放蛋白(MRP)对构成血脑屏 障的微血管内皮细胞有何致病作用迄今不明。为此分离仔兔脾微血管内皮细胞（SMEC），纯化后用SV40-T抗原 转化后用作试验的细胞模型。将单层SMEC 和电泳纯MRP 溶液共孵育，染色后，光镜观察，发现MRP可诱导SMEC 发生两种典型形态学变化：致密细胞单层中出现巨大空洞而呈网状；空洞内有细胞融合，形成多核巨细胞，随 后巨细胞核极度浓缩释出，巨细胞消失。研究结果表明， MRP单独足以破坏大脑的血管内皮细胞屏障。 关键词：猪链球菌2型；溶菌酶释放蛋白；细胞融合；细胞凋亡；血管内皮细胞 Fusion of Microvascular Endothelial Cells Induced by Muramidase-Released Protein of Streptococcus suis Type 2 ZENG Qiao-ying 1, 2 ，LU Cheng-ping 1 1 ( Key Laboratory of Animal Disease Diagnostic and Immunology, Ministry of Agriculture, Nanjing Agricultural University, 2 Nanjing 210095； College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070 ) Abstract: Streptococcus suis type 2 (SS2) mainly causes meningitis of human and pigs. Muramidase-released protein (MRP) is one of the key virulence factors of SS2. However ，the pathogenic effects of MRP on microvascular endothelial cells (MEC) which makes up of brain blood barrier (BBB) is unknown to date. So the spleen microvascular endothelial cells (SMEC) from neonatal rabbit were isolated, purified, immortalized by SV40-T antigen, and used as cell model in present study. After co-incubation with electrophoretically purified MRP, two principal morphologic changes of SMEC confluent layer were revealed by light microscopy. Firstly, the confluent cell layer turned into a riddle-like stru