Identification of apoptosis and macrophage migration events in paraquat-induced oxidative stress using a zebrafish model.
Identification of apoptosis and macrophage migration events in paraquat-induced oxidative stress using a zebrafish model.
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DOI：
10.1016/j.lfs.2016.06.009
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发表时间：
2016-07
期刊：
影响因子：
6.1
通讯作者：
Qiang Wang;Shuai Liu;Dayong Hu;Zhen Wang;Ling Wang;Tianfu Wu;Zhuanbin Wu;C. Mohan;Ai Peng
中科院分区：
文献类型：
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作者：
Qiang Wang;Shuai Liu;Dayong Hu;Zhen Wang;Ling Wang;Tianfu Wu;Zhuanbin Wu;C. Mohan;Ai Peng
百草枯（Paraquat，PQ）是一种对人体有很高毒性的农药，也是一种众所周知的氧化应激触发剂。虽然已经建立了几种PQ中毒的动物模型，但仍有一些缺点限制了它们在体内的应用。在本研究中使用的斑马鱼模型，以更好地确定由PQ引起的氧化应激损伤的机制。主要方法PQ的毒性进行了评估，在斑马鱼AB株，并通过吖啶橙子染色的细胞凋亡进行了评估。使用TG（zlyz：EGFP）转基因株鉴定巨噬细胞迁移，并使用fli 1a-EGFP casper株观察血管生成。在对斑马鱼体内实时荧光定量PCR基因变化进行验证后，利用Influencity Pathway Analysis软件进行网络分析。关键发现我们首先在斑马鱼模型中建立了PQ的LC 50，然后发现PQ暴露后鲁棒的氧化应激和抗氧化基因被激活。此外，细胞凋亡和不同的巨噬细胞活化和迁移首次在PQ暴露的斑马鱼。利用这个模型，外源性和内源性途径参与PQ诱导的细胞凋亡被阐明。我们进一步证明PQ特异性地诱导巨噬细胞迁移，Rho家族成员和JNK-MMP 13信号通路参与了这一过程。
AimsParaquat (PQ) is a pesticide highly toxic to human beings, and a well-known trigger of oxidative stress. Although several animal models of PQ poisoning have been developed, some disadvantages limit their application in vivo. A zebrafish model was used in the present study to better define mechanisms of oxidative stress injury induced by PQ.Main methodsThe toxicity of PQ was evaluated in the AB strain of zebrafish, and apoptosis was assessed by acridine orange staining. Macrophage migration was identified using the TG (zlyz:EGFP) transgenic strain, and angiogenesis was observed using thefli1a-EGFP casperstrain. Following the validation of gene changes by zebrafish-based in vivo quantitative real-time PCR, network analysis was performed using the Ingenuity Pathway Analysis software.Key findingsWe first established the LC50of PQ in the zebrafish model, and then found that robust oxidative stress and antioxidant genes were activated after PQ exposure. Moreover, apoptosis and distinct macrophage activation and migration were identified for the first time in PQ-exposed zebrafish. Utilizing this model, both extrinsic and intrinsic pathways involved in PQ-induced apoptosis were elucidated. We further demonstrated that macrophage migration was specifically induced by PQ, and that Rho family members and JNK-MMP13 signaling participated in this process.SignificanceZebrafish is a promising tool for investigating the mechanisms of oxidative stress injury induced by PQ, and for screening effective anti-oxidant drugs.