Porcine hemagglutinating encephalomyelitis virus induces apoptosis. in a porcine kidney cell line via caspase-dependent pathways
Porcine hemagglutinating encephalomyelitis virus induces apoptosis. in a porcine kidney cell line via caspase-dependent pathways
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猪血凝脑脊髓炎病毒通过半胱天冬酶依赖性途径诱导猪肾细胞系凋亡
DOI：
10.1016/j.virusres.2013.05.019
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发表时间：
2013-09-01
期刊：
影响因子：
5
通讯作者：
He, Wenqi
中科院分区：
文献类型：
--
作者：
Lan, Yungang;Zhao, Kui;He, Wenqi
猪血凝性脑脊髓炎是由猪血凝性脑脊髓炎病毒（PHEV）引起的仔猪急性高度接触性传染病。然而，PHEV的发病机制以及PHEV与宿主细胞的关系尚未完全清楚。在这项研究中，我们研究了PHEV诱导的细胞病变效应（CPE）是否是由细胞凋亡引起的。PHEV在猪肾衍生细胞系（PK-15细胞）中的复制引起广泛的CPE，导致整个单层的破坏和感染细胞的死亡。Hoechst 33，342染色显示细胞核和染色质碎片的形态学变化。此外，PHEV引起的DNA片段检测琼脂糖凝胶电泳感染后48小时，随着孵育时间的增加。凋亡细胞的百分比随着孵育时间的增加而增加，并在感染后96 h达到最大值，如使用流式细胞术和荧光显微镜用膜联蛋白V-FITC和碘化丙啶（PI）染色的细胞所确定的。此外，正如在其他动物的冠状病毒感染中通常观察到的那样，在PHEV感染的PK-15细胞中，分别作为死亡受体介导的凋亡途径和线粒体凋亡途径中的代表性因子的效应物半胱天冬酶（caspase）、半胱天冬酶-3和起始物半胱天冬酶（caspase）、半胱天冬酶-8和半胱天冬酶-9的活性增加。此外，三肽pan-ICE（半胱天冬酶）抑制剂Z-VAD-FMK阻断PHEV诱导的细胞凋亡，但在感染后96小时对病毒产生没有影响。这些结果表明，PHEV诱导PK-15细胞凋亡通过半胱天冬酶依赖的途径。感染细胞的凋亡死亡对动物是有害的，因为它会导致细胞和组织的破坏。尽管PHEV的病理学特征尚不清楚，但细胞凋亡可能是PHEV感染引起病变的病理学基础。(C)2013爱思唯尔有限公司版权所有。
Porcine hemagglutinating encephalomyelitis is an acute, highly contagious disease in piglets that is caused by the porcine hemagglutinating encephalomyelitis virus (PHEV). However, the pathogenesis of PHEV and the relationship between PHEV and the host cells are not fully understood. In this study, we investigated whether the PHEV-induced cytopathic effect (CPE) was caused by apoptosis. Replication of PHEV in a porcine kidney-derived cell line (PK-15 cells) caused an extensive CPE, leading to the destruction of the entire monolayer and the death of the infected cells. Staining with Hoechst 33,342 revealed morphological changes in the nuclei and chromatin fragmentation. In addition, PHEV caused DNA fragmentation detectable by agarose gel electrophoresis 48 h post-infection, increasing with the incubation time. The percentage of apoptotic cells increased with the incubation time and reached a maximum at 96 h post-infection, as determined using flow cytometry and fluorescence microscopy of cells that were stained with annexin V-FITC and propidium iodide (PI). Moreover, as is commonly observed for coronavirus infections of other animals, the activities of the effecter caspase, caspase-3, and the initiator caspases, caspase-8 and caspase-9, which are representative factors in the death receptor-mediated apoptotic pathway and the mitochondrial apoptotic pathway, respectively, were increased in PHEV-infected PK-15 cells. Moreover, the tripeptide pan-ICE (caspase) inhibitor Z-VAD-FMK blocked PHEV-induced apoptosis but did not have an effect on virus production by 96 h post-infection. These-results suggested that PHEV induces apoptosis in PK-15 cells via a caspase-dependent pathway. Apoptotic death of infected cells is detrimental to animals because it causes cell and tissue destruction. Although the pathological characteristics of PHEV are largely unknown, apoptosis may be the pathological basis of the lesions resulting from PHEV infection. (C) 2013 Elsevier B.V. All rights reserved.