2型登革病毒通过线粒体途径诱导EA. hy926细胞凋亡

发布时间：2018-01-04 20:28

本文关键词：2型登革病毒通过线粒体途径诱导EA. hy926细胞凋亡　出处：《中国病理生理杂志》2013年03期 　论文类型：期刊论文

【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(P0.05,P0.01);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(P0.01)。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。
[Abstract]:Objective: To investigate the dengue virus induced EA.hy926 cells (cell line of human umbilical vein endothelial cell fusion) and changes of mitochondrial membrane potential (mitochondrial membrane relative activity of potential, m and delta psi) relationship between the changes of mitochondrial apoptosis pathway. Methods: dengue virus type 2 (denguevirus type 2, DENV-2) infection of EA.hy926 cells, MTT method to detect the infection of EA.hy926 cells before and after the relative activity, fluorescence microscopy and flow cytometry were used to observe the aggregation of JC-1 before and after infection in EA.hy926 cell mitochondria to detect the change of M delta psi, by changing the activity of the caspase-9 colorimetric assay. Results: the infection of DENV-2 EA hy926 cells in 24 h, 36 h and 48 h after the cell activity was inhibited at 550 nm A values were lower than the uninfected group, the difference was statistically significant (P0.05, P0.01); JC-1 staining showed red fluorescence at different time points after infection, on behalf of normal mitochondria were Compared with the uninfected group decreased, and the representative of green fluorescent m decreased compared with delta psi uninfected group increased gradually. The mean fluorescence density of delta psi detection m than the uninfected group. Flow cytometry, differences can occur early caspase-9 activity increased statistical significance after.DENV-2 infection, compared with uninfected group, activity difference at each time point there was statistically significant (P0.01). Conclusion: DENV-2 EA infection, hy926 cells can induce Delta Psi m decreased, increased caspase-9 activity and apoptosis of mitochondrial pathway.

【作者单位】：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;
【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)
【分类号】：R373.33
【正文快照】： 登革热是是世界上最常见的蚊媒传播疾病之一,但目前仍缺乏合适的实验动物模型和特异性的治疗手段,也没有安全的疫苗可以预防其传播。对于重症型的登革出血热(dengue hemorrhagic fever,DHF)或登革休克综合征(dengue shock syndrome,DSS),血管内皮损伤导致的血浆渗漏及出血是

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