Li Su, Qu Hui, Hao Jianwei, Sun Jinfu, Guo Huancheng, Guo Changming, Sun Boxing, Tu Changchun
College of Animal Science and Veterinary Medicine, Jilin University, 5333 XiAn Road, Changchun 130062, China.
Biochim Biophys Acta. 2010 Sep;1804(9):1882-8. doi: 10.1016/j.bbapap.2010.05.011. Epub 2010 Jun 1.
Endothelial cells are the main target of classical swine fever virus during infection, and extensive hemorrhage is the most typical clinical sign of classical swine fever. To investigate the molecular mechanism of hemorrhagic pathogenesis, two-dimensional difference gel electrophoresis with fluorescent dyes (2D-DIGE) was used to analyze the proteomic profile of primary porcine umbilical vein endothelial cells (PUVECs) following CSFV infection. Of 15 protein spots with differential expression, 8 were characterized by MALDI-TOF-MS/MS in infected PUVECs at 48h p.i.: moesin, peroxiredoxin 6, stathmin-1, a protein similar to nascent polypeptide-associated complex alpha subunit isoform 2, phosphoglycerate kinase 1, glucosidase II, transketolase and alpha-tubulin. These could be sorted into 5 functional groups: glycometabolism, cell proliferation, anti-oxidative stress, inflammatory response and cytoskeleton. Western blot and real-time RT-PCR analysis confirmed the down-regulation of phosphoglycerate kinase 1 (PGK1) and up-regulation of moesin identified by 2D-DIGE. Pathway analysis of these 15 differentially expressed proteins showed that CSFV infection altered the metabolism, cytoskeleton and cell proliferation of PUVECs, and that consequently an inflammatory response was induced.
内皮细胞是猪瘟病毒感染过程中的主要靶细胞，广泛出血是猪瘟最典型的临床症状。为了研究出血性发病机制的分子机制，采用荧光染料双向差异凝胶电泳（2D-DIGE）分析猪瘟病毒感染后原代猪脐静脉内皮细胞（PUVECs）的蛋白质组图谱。在15个差异表达的蛋白点中，通过基质辅助激光解吸电离飞行时间串联质谱（MALDI-TOF-MS/MS）对感染后48小时的PUVECs中的8个蛋白进行了鉴定：肌动蛋白结合蛋白、过氧化物酶体增殖物激活受体6、微管相关蛋白1、一种与新生多肽相关复合物α亚基异构体2相似的蛋白、磷酸甘油酸激酶1、葡萄糖苷酶II、转酮醇酶和α-微管蛋白。这些蛋白可分为5个功能组：糖代谢、细胞增殖、抗氧化应激、炎症反应和细胞骨架。蛋白质免疫印迹和实时逆转录聚合酶链反应分析证实了2D-DIGE鉴定的磷酸甘油酸激酶1（PGK1）的下调和肌动蛋白结合蛋白的上调。对这15个差异表达蛋白的通路分析表明，猪瘟病毒感染改变了PUVECs的代谢、细胞骨架和细胞增殖，从而诱导了炎症反应。
