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Duck Plague Virus Promotes DEF Cell Apoptosis by Activating Caspases, Increasing Intracellular ROS Levels and Inducing Cell Cycle S-Phase Arrest
Background: Duck plague virus (DPV) can induce apoptosis in duck embryo fibroblasts (DEFs) and in infected ducks, but the molecular mechanism of DPV-induced apoptosis remains unknown. Methods: We first used qRT-PCR and a Caspase-Glo assay to determine whether the caspase protein family plays an impo...
Autores principales: | , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6409732/ https://www.ncbi.nlm.nih.gov/pubmed/30813500 http://dx.doi.org/10.3390/v11020196 |
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author | Zhao, Chuankuo Wang, Mingshu Cheng, Anchun Yang, Qiao Wu, Ying Jia, Renyong Zhu, Dekang Chen, Shun Liu, Mafeng Zhao, Xinxin Zhang, Shaqiu Liu, Yunya Yu, Yanling Zhang, Ling Tian, Bin Rehman, Mujeeb Ur Pan, Leichang Chen, Xiaoyue |
author_facet | Zhao, Chuankuo Wang, Mingshu Cheng, Anchun Yang, Qiao Wu, Ying Jia, Renyong Zhu, Dekang Chen, Shun Liu, Mafeng Zhao, Xinxin Zhang, Shaqiu Liu, Yunya Yu, Yanling Zhang, Ling Tian, Bin Rehman, Mujeeb Ur Pan, Leichang Chen, Xiaoyue |
author_sort | Zhao, Chuankuo |
collection | PubMed |
description | Background: Duck plague virus (DPV) can induce apoptosis in duck embryo fibroblasts (DEFs) and in infected ducks, but the molecular mechanism of DPV-induced apoptosis remains unknown. Methods: We first used qRT-PCR and a Caspase-Glo assay to determine whether the caspase protein family plays an important role in DPV-induced apoptosis. Then, we used an intracellular ROS detection kit and the mitochondrial probe JC-1 to respectively detect ROS levels and mitochondrial membrane potential (MMP). Finally, flow cytometry was used to detect apoptosis and cell cycle progression. Results: In this study, the mRNA levels and enzymatic activities of caspase-3, caspase-7, caspase-8, and caspase-9 were significantly increased during DPV-induced apoptosis. The caspase inhibitors Z-DEVD-FMK, Z-LEHD-FMK, and Q-VD-Oph could inhibit DPV-induced apoptosis and promote viral replication. Subsequently, a significant decrease in MMP and an increase in the intracellular ROS levels were observed. Further study showed that pretreating infected cells with NAC (a ROS scavenger) decreased the intracellular ROS levels, increased the MMP, inhibited apoptosis, and promoted viral replication. Finally, we showed that DPV infection can cause cell cycle S-phase arrest. Conclusions: This study shows that DPV causes cell cycle S-phase arrest and leads to apoptosis through caspase activation and increased intracellular ROS levels. These findings may be useful for gaining an understanding of the pathogenesis of DPV and the apoptotic pathways induced by α-herpesviruses. |
format | Online Article Text |
id | pubmed-6409732 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-64097322019-04-01 Duck Plague Virus Promotes DEF Cell Apoptosis by Activating Caspases, Increasing Intracellular ROS Levels and Inducing Cell Cycle S-Phase Arrest Zhao, Chuankuo Wang, Mingshu Cheng, Anchun Yang, Qiao Wu, Ying Jia, Renyong Zhu, Dekang Chen, Shun Liu, Mafeng Zhao, Xinxin Zhang, Shaqiu Liu, Yunya Yu, Yanling Zhang, Ling Tian, Bin Rehman, Mujeeb Ur Pan, Leichang Chen, Xiaoyue Viruses Article Background: Duck plague virus (DPV) can induce apoptosis in duck embryo fibroblasts (DEFs) and in infected ducks, but the molecular mechanism of DPV-induced apoptosis remains unknown. Methods: We first used qRT-PCR and a Caspase-Glo assay to determine whether the caspase protein family plays an important role in DPV-induced apoptosis. Then, we used an intracellular ROS detection kit and the mitochondrial probe JC-1 to respectively detect ROS levels and mitochondrial membrane potential (MMP). Finally, flow cytometry was used to detect apoptosis and cell cycle progression. Results: In this study, the mRNA levels and enzymatic activities of caspase-3, caspase-7, caspase-8, and caspase-9 were significantly increased during DPV-induced apoptosis. The caspase inhibitors Z-DEVD-FMK, Z-LEHD-FMK, and Q-VD-Oph could inhibit DPV-induced apoptosis and promote viral replication. Subsequently, a significant decrease in MMP and an increase in the intracellular ROS levels were observed. Further study showed that pretreating infected cells with NAC (a ROS scavenger) decreased the intracellular ROS levels, increased the MMP, inhibited apoptosis, and promoted viral replication. Finally, we showed that DPV infection can cause cell cycle S-phase arrest. Conclusions: This study shows that DPV causes cell cycle S-phase arrest and leads to apoptosis through caspase activation and increased intracellular ROS levels. These findings may be useful for gaining an understanding of the pathogenesis of DPV and the apoptotic pathways induced by α-herpesviruses. MDPI 2019-02-24 /pmc/articles/PMC6409732/ /pubmed/30813500 http://dx.doi.org/10.3390/v11020196 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Zhao, Chuankuo Wang, Mingshu Cheng, Anchun Yang, Qiao Wu, Ying Jia, Renyong Zhu, Dekang Chen, Shun Liu, Mafeng Zhao, Xinxin Zhang, Shaqiu Liu, Yunya Yu, Yanling Zhang, Ling Tian, Bin Rehman, Mujeeb Ur Pan, Leichang Chen, Xiaoyue Duck Plague Virus Promotes DEF Cell Apoptosis by Activating Caspases, Increasing Intracellular ROS Levels and Inducing Cell Cycle S-Phase Arrest |
title | Duck Plague Virus Promotes DEF Cell Apoptosis by Activating Caspases, Increasing Intracellular ROS Levels and Inducing Cell Cycle S-Phase Arrest |
title_full | Duck Plague Virus Promotes DEF Cell Apoptosis by Activating Caspases, Increasing Intracellular ROS Levels and Inducing Cell Cycle S-Phase Arrest |
title_fullStr | Duck Plague Virus Promotes DEF Cell Apoptosis by Activating Caspases, Increasing Intracellular ROS Levels and Inducing Cell Cycle S-Phase Arrest |
title_full_unstemmed | Duck Plague Virus Promotes DEF Cell Apoptosis by Activating Caspases, Increasing Intracellular ROS Levels and Inducing Cell Cycle S-Phase Arrest |
title_short | Duck Plague Virus Promotes DEF Cell Apoptosis by Activating Caspases, Increasing Intracellular ROS Levels and Inducing Cell Cycle S-Phase Arrest |
title_sort | duck plague virus promotes def cell apoptosis by activating caspases, increasing intracellular ros levels and inducing cell cycle s-phase arrest |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6409732/ https://www.ncbi.nlm.nih.gov/pubmed/30813500 http://dx.doi.org/10.3390/v11020196 |
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