中国免疫学杂志2016，Vol.32Issue(7)：945-951,7.DOI:10.3969/j.issn.1000-484X.2016.07.003
登革病毒Ⅱ型引起原代HDMECs通透性变化机制的初步研究
Primary mechanism of changing permeability in DENV-2 infected primary human dermal micro-vascular endothelial cells
摘要
Abstract
Objective:To reveal the primary mechanism of changing permeability in DENV-2 infected pHDMECs. Methods:pHDMECs was incubated by DENV-2 on the concentration of 103 TCID50 ,and the penetrability of the cell was detected by Transwell at 4,8,12,24,48 h,respectively. Then,the partial sequence of DENV-2 NS1 was analyzed by Real time-PCR,and NS1 protein was detected by immunofluorescence and flow cytometer (FCM). The apoptosis rate of pHDMECs was assayed by FCM. Finally,IL-6 and IL-8 secreted by pHDMECs were analyzed by Real time-PCR and double antibody sandwich ELISA. Results:The relative expression of NS1 gene was elevated but NS1 protein was not detected;the permeability of DENV-2 infected pHDMECs had dramatically increased both at 24,48 h,but the apoptosis rate has little changed even been influenced by DENV-2 at 72 h. However,the relative expression of IL-6/IL-8 mRNA was boosted at 8,24 h[(2. 49±0. 50) and (6. 82±1. 69) fold,respectively,P<0. 05]. In protein level,compared with control(869. 6±50. 70)pg/ml,IL-6 secreted by DENV-2 infected pHDMECs could reach by(1 248. 8±86. 9)pg/ml(P<0. 05),and IL-8 was(1 331. 0±86. 3)pg/ml(P<0. 05) while the control was (967. 6±156. 6)pg/ml. Conclusion:Indeed,pHDMECs can be infected by DENV-2;the increasing permeability of DENV-2 infected pHDMECs may not be caused by the pHDMECs′ apoptosis but the enhancing of pro-inflammatory cytokine IL-6 /IL-8.关键词
登革病毒/原代人真皮微血管内皮细胞/IL-6/IL-8/通透性Key words
Dengue virus/Primary human dermal micro-vascular endothelial cells/IL-6/IL-8/Permeability分类
基础医学引用本文复制引用
赵军,左丽,戴雪婷,裴华,袁静,孔维莹..登革病毒Ⅱ型引起原代HDMECs通透性变化机制的初步研究[J].中国免疫学杂志,2016,32(7):945-951,7.基金项目
本文为国家自然科学基金(31260224 （）
81560263)和贵州省教育厅“125”重大科技专项(黔教合重大专项字[2012]008号)。 （黔教合重大专项字[2012]008号）
