Porcine parvovirus infection activates mitochondria-mediated apoptotic signaling pathway by inducing ROS accumulation.
Porcine parvovirus infection activates mitochondria-mediated apoptotic signaling pathway by inducing ROS accumulation.
复制标题
猪细小病毒感染通过诱导ROS积累激活线粒体介导的细胞凋亡信号通路
DOI：
10.1186/s12985-016-0480-z
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发表时间：
2016-02-16
期刊：
影响因子：
4.8
通讯作者：
Tong D
中科院分区：
文献类型：
--
作者：
Zhao X;Xiang H;Bai X;Fei N;Huang Y;Song X;Zhang H;Zhang L;Tong D
背景猪细小病毒（PPV）感染主要导致妊娠猪繁殖失败并导致宿主细胞死亡。公猪作为重要的传播者，通过精液将PPV传播给母猪。 PPV感染公猪睾丸并在其体内大量复制，导致猪睾丸体内损伤。活性氧（ROS）是细胞凋亡的介质，在线粒体凋亡途径中发挥着至关重要的作用。然而，PPV感染是否会诱导ST细胞凋亡和ROS积累尚不清楚。方法为了确定PPV感染对细胞凋亡的影响，我们检测PPV感染的ST细胞的形态变化、DNA梯子、caspases活性和PARP表达。此外，为了研究PPV感染对线粒体凋亡途径和ROS积累的影响，我们检测了Δψm、凋亡相关基因和ROS。为了研究ROS在PPV诱导的细胞凋亡过程中的作用，将ST细胞感染PPV并用ROS抗氧化剂处理。使用活性氧检测试剂盒测量ROS水平，并测试Δψm、Bcl-2的表达水平、Bax的易位和线粒体细胞色素c的重新分布。 结果在本研究中，我们证明PPV感染可以诱导细胞凋亡，其特征是形态变化、DNA片段化和caspases激活。此外，PPV感染抑制Bcl-2表达，增强Bax表达和向线粒体的易位，降低线粒体跨膜电位，并触发细胞色素c的释放，从而引起随后的caspase-9和caspase-3激活并启动细胞凋亡。然而，在PPV诱导细胞凋亡的过程中，Fas和FasL的蛋白水平并未受到影响。进一步的研究表明PPV感染引起ROS积累。抑制ROS可以降低线粒体跨膜电位，并通过抑制Bax易位、细胞色素c和caspase-3活化来显着阻断ST细胞凋亡。结论所有这些结果表明PPV诱导的ROS积累介导了ST细胞的凋亡，这为PPV感染的分子发病机制提供了理论依据。
BackgroundPorcine parvovirus (PPV) infection primarily causes reproductive failure of pregnant swine and results in host cell death. Boars, as an important disseminator, shed PPV to sows via semen. PPV infects and numerously replicates in boar testicle, which results in damage of swine testicle in vivo. Reactive oxygen species (ROS), a mediator of cell apoptosis, play a crucial role in the mitochondria apoptotic pathway. However, whether PPV infection induces ST cells apoptosis and ROS accumulation is still unclear.MethodsTo determine the effects of PPV infection on the apoptosis, we detected morphological changes, DNA ladder, activities of caspases, and expression of PARP in PPV-infected ST cells. Moreover, aiming to investigate the effect of PPV infection on the mitochondrial apoptotic pathway and ROS accumulation, we detected the Δψm, apoptosis-related genes, and ROS. To investigate the role of ROS in the process of PPV-induced apoptosis, the ST cells were infected with PPV and treated with the ROS antioxidants. The ROS level was measured using Reactive Oxygen Species Assay Kit and the Δψm, expression level of Bcl-2, translocation of Bax, and redistribution of mitochondria cytochrome c were tested.ResultsIn this study, we demonstrated that PPV infection could induce apoptosis that was characterized by morphological changes, DNA fragmentation and activation of caspases. Moreover, PPV infection suppressed Bcl-2 expression, enhanced Bax expression and translocation to mitochondria, decreased the mitochondrial transmembrane potential, and triggered the release of cytochrome c, which caused the subsequent activation of caspase-9 and caspase-3 and initiation of apoptosis. However, during the process of PPV-induced apoptosis, the protein levels of Fas and FasL were not affected. Further studies showed that PPV infection caused ROS accumulation. Inhibition of ROS could reduce mitochondrial transmembrane potential and could significantly block ST cells apoptosis via suppressing Bax translocation, cytochrome c and caspase-3 activation.ConclusionsAll these results suggest that PPV-induced ROS accumulation mediates apoptosis in ST cells, which provided theoretical basis for the molecular pathogenesis of PPV infection.