2型登革病毒通过线粒体途径诱导EA．hy926细胞凋亡  

DENV-2 induces apoptosis of EA. hy926 cells through mitochondrial pathway

作　　者：齐一鸣[1] 黄俊琪[1] 

机构地区：[1]中山大学中山医学院免疫学研究所,教育部热带病防治研究重点实验室,广东广州510080

出　　处：《中国病理生理杂志》2013年第3期385-389,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30872350);广东省自然科学基金资助项目(No.S2012010009050);广东省科技计划项目(No.2010B050700008;No.2011B040300022);广州市科技计划项目(No.2011J4100084)

摘　　要：目的:探讨登革病毒诱导EA.hy926细胞(人脐静脉内皮细胞融合细胞株)相对活力的变化与线粒体膜电位(mitochondrial membrane potential,Δψm)改变及线粒体凋亡途径的关系。方法:用2型登革病毒(denguevirus type 2,DENV-2)感染EA.hy926细胞,MTT法检测感染前后EA.hy926细胞的相对活力,荧光显微镜和流式细胞术分别观察感染前后JC-1在EA.hy926细胞线粒体内的聚集情况以检测Δψm的改变,通过比色法检测caspase-9的活性变化。结果:DENV-2感染EA·hy926细胞24 h、36 h及48 h后,细胞活性受到显著抑制,550 nm处的A值均低于未感染组,差异有统计学意义(P<0.05,P<0.01);JC-1染色显示,感染后各时点,代表正常线粒体的红色荧光均较未感染组减弱,而代表Δψm下降的绿色荧光较未感染组逐渐增强。流式细胞术检测Δψm平均荧光密度比未感染组减低,差异有统计学意义。DENV-2感染后早期即可出现caspase-9活性的上升,与未感染组相比,各时点的活性差异均有统计学意义(P<0.01)。结论:DENV-2感染EA·hy926细胞后可诱发Δψm下降,增强caspase-9活性,进而启动线粒体的凋亡途径。AIM ： To explore the effect of dengue virus type 2 （DENV-2） infection on the change of mitochon- drial membrane potential （A~,m ） in EA. hy926 cells. METHODS： The inhibitory effect of DENV-2 infection on EA. hy926 cell growth was examined by MTr assay. The changes of Δψm were analyzed by flow cytometry or observed under flu- orescence microscope with JC-1 staining. The activity of caspase-9 was measured by a colorimetric kit. RESULTS ： Infec- tion of DENV-2 for 24 h, 36 h and 48 h inhibited the viability of EA. hy926 cells. After DENV-2 infection, the changes of Δψm, in EA. hy926 ceils were observed. Compared with the normal control ceils, Δψm in DENV-2-infected EA. hy926 cells was notably decreased. The activity of caspase-9 increased at early stage after infection of DENV-2 and maintained at a high level at least to 48 h. CONCLUSION： DENV-2 infection decreases the mitochondrial membrane potential and increases the activity of caspase-9 in EA. hy926 cells in the early stage of proliferation, thus promoting the process of apoptosis.

关 键 词：登革病毒 线粒体膜电位 血管内皮细胞 

分 类 号：R373.33[医药卫生—病原生物学]