Zhang Li, Li Ling, Wang Bin, Qian Dong-Meng, Song Xu-Mia, Hu Ming
Department of Microbiology, Key Laboratory of Medicine and Biotechnology of Qingdao, Qingdao University Medical College, Qingdao, Shandong, China.
Neuroreport. 2013 Jul 10;24(10):526-35. doi: 10.1097/WNR.0b013e32836206d1.
Transmission of human cytomegalovirus (HCMV) to the fetus is the most common type of intrauterine infection; the mechanism of HCMV pathogenesis in the developing central nervous system remains unclear. Thrombospondins 1 and 2 (TSP1, TSP2) produced by immature astrocytes are critical for fetal synaptogenesis. To examine the effect of HCMV on fetal astrocytes, human fetal astrocytes were isolated and cultured with HCMV AD169. Cells were harvested at different time points. Protein and mRNA expressions of TSP1 and TSP2 were determined using RT-qPCR, western blotting analysis, and enzyme-linked immunosorbent assay. The results showed that HCMV infection induced time-dependent decreases in mRNA and protein expressions of both TSP1 and TSP2 in astrocytes. Flow cytometry was used to detect apoptosis of HCMV-infected astrocytes, and the result indicated that there was no linkage between cell apoptosis and the decrease in TSP1 and TSP2 expressions induced by HCMV infection. When ganciclovir treatment was performed on HCMV-infected astrocytes, results showed that ganciclovir treatment inhibited the reduction of TSP1 and TSP2 expression in astrocytes. In the further study, pEGFP-N3-IE1 was transfected into astrocytes to identify that it was not IE1 but active viral replication that was essential in the continuous decrease of TSP1 and TSP2 expressions in HCMV-infected astrocytes.
人巨细胞病毒（HCMV）传播给胎儿是最常见的宫内感染类型；HCMV在发育中的中枢神经系统发病机制仍不清楚。未成熟星形胶质细胞产生的血小板反应蛋白1和2（TSP1、TSP2）对胎儿突触形成至关重要。为了研究HCMV对胎儿星形胶质细胞的影响，分离人胎儿星形胶质细胞并用HCMV AD169进行培养。在不同时间点收获细胞。使用RT-qPCR、蛋白质印迹分析和酶联免疫吸附测定法测定TSP1和TSP2的蛋白质和mRNA表达。结果表明，HCMV感染诱导星形胶质细胞中TSP1和TSP2的mRNA和蛋白质表达呈时间依赖性下降。使用流式细胞术检测HCMV感染的星形胶质细胞的凋亡，结果表明细胞凋亡与HCMV感染诱导的TSP1和TSP2表达下降之间没有关联。当对HCMV感染的星形胶质细胞进行更昔洛韦处理时，结果表明更昔洛韦处理抑制了星形胶质细胞中TSP1和TSP2表达的降低。在进一步的研究中，将pEGFP-N3-IE1转染到星形胶质细胞中，以确定在HCMV感染的星形胶质细胞中，TSP1和TSP2表达持续下降的关键不是IE1，而是活跃的病毒复制。
