南方医科大学学报 ›› 2015, Vol. 35 ›› Issue (06): 857-.

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肠道病毒71型对Vero细胞线粒体的损伤作用及机制

林佩欣,申 洪,高璐璐,黄业恩,张耀忠,陈 清   

  • 出版日期:2015-06-20 发布日期:2015-06-20

Mitochondrial injury in enterovirus 71-infected Vero cells and its mechanism

  • Online:2015-06-20 Published:2015-06-20

摘要: 目的探讨肠道病毒71型(EV71)对Vero细胞的损伤作用及机制,为分析EV71的致病机制提供新思路。方法将临床分
离的一株EV71接种于Vero细胞,观察细胞的形态改变,用免疫荧光方法检测EV71抗原在Vero细胞中的表达,用透射电镜观察
Vero细胞超微结构的改变,用Image-Pro Plus 6.0图像分析软件测试分析病毒颗粒直径和面积密度以及空泡化线粒体的比例和
面积密度。结果EV71感染后Vero细胞形态发生改变,由梭形变为圆形或死亡,免疫荧光检测显示EV71抗原定位于胞质,超
微结构观察可见胞质中有大量呈团块状分布、晶格状排列的高电子密度病毒颗粒,颗粒平均直径为16.3 nm,平均面积密度为
38.3%;线粒体肿胀、变性并空泡化、崩解,空泡化线粒体数量约占90.9%,平均面积密度为89.2%;部分线粒体内可见EV71病毒
颗粒。结论EV71感染Vero细胞后在其胞质中增殖,并可侵入线粒体,导致线粒体直接损伤并致细胞死亡;线粒体是EV71在
Vero细胞内的作用靶标。

Abstract: Objective To investigate the mitochondrial injury in enterovirus 71 (EV71)-infected Vero cells and explore the
possible mechanism. Methods A clinical isolate of EV71 was inoculated to Vero cells and the EV71 antigen was detected by
immunofluorescence assay. The morphological changes of Vero cells were observed using optical microscopy and transmission
electron microscopy. The diameter and area density of the viral particles and the ratio and area density of vacuolated
mitochondria in the cells were measured on the ultrastructural images. Results EV71-infected Vero cells underwent obvious
changes and to a spherical morphology followed by cell death EV71 particles were detected in the cytoplasm by
immunofluorescence. Ultrastructurally, the infected cells contained a large number of viral particles in the cytoplasm, with a
clustered distribution and lattice-like arrangement. The diameter of the particles were 16.3 nm and the mean area density was
38.3% . Most of the mitochondria presented with swelling, vacuoles and degeneration. The ratio of the vacuolated
mitochondria was 90.9% with a mean area density of 89.2%. Viral particles were also found in some mitochondria. Conclusion
EV71 proliferates in the cytoplasm and invades the mitochondria of infected Vero cells leading to mitochondrial injury and cell
death, suggesting that mitochondria are the targets for EV71 infection.