Primary mechanism of changing permeability in DENV-2 infected primary human dermal micro-vascular endothelial cells
10.3969/j.issn.1000-484X.2016.07.003
- VernacularTitle:登革病毒Ⅱ型引起原代HDMECs通透性变化机制的初步研究
- Author:
Jun ZHAO
;
Li ZUO
;
Xueting DAI
;
Hua PEI
;
Jing YUAN
;
Weiying KONG
- Publication Type:Journal Article
- Keywords:
Dengue virus;
Primary human dermal micro-vascular endothelial cells;
IL-6;
IL-8;
Permeability
- From:
Chinese Journal of Immunology
2016;32(7):945-951
- CountryChina
- Language:Chinese
-
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.
