CAJ | 학술논문

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。
목적：탐토장도병독71형（EV71）대Vero세포적손상작용급궤제，위분석EV71적치병궤제제공신사로。방법장림상분리적일주EV71접충우Vero세포，관찰세포적형태개변，용면역형광방법검측EV71항원재Vero세포중적표체，용투사전경관찰Vero세포초미결구적개변，용Image-Pro Plus 6.0도상분석연건측시분석병독과립직경화면적밀도이급공포화선립체적비례화면적밀도。결과 EV71감염후Vero세포형태발생개변，유사형변위원형혹사망，면역형광검측현시EV71항원정위우포질，초미결구관찰가견포질중유대량정단괴상분포、정격상배렬적고전자밀도병독과립，과립평균직경위16.3 nm，평균면적밀도위38.3%；선립체종창、변성병공포화、붕해，공포화선립체수량약점90.9%，평균면적밀도위89.2%；부분선립체내가견EV71병독과립。결론 EV71감염Vero세포후재기포질중증식，병가침입선립체，도치선립체직접손상병치세포사망；선립체시EV71재Vero세포내적작용파표。
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.