Pro-inflammatory regulatory mechanism of C5a/C5aR pathway in GCRV-infected grass carp (Ctenopharyngodon idella)
Pro-inflammatory regulatory mechanism of C5a/C5aR pathway in GCRV-infected grass carp (Ctenopharyngodon idella)
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DOI：
10.1016/j.aqrep.2023.101736
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发表时间：
2023-09-15
影响因子：
3.7
通讯作者：
Wen，Zheyu
中科院分区：
文献类型：
--
作者：
Xu，Baohong;Su，Hang;Wen，Zheyu
草鱼呼肠孤病毒（GCRV）感染引起草鱼大量死亡，严重制约了草鱼养殖业的发展。补体系统中C5 a和C5 a受体（C5 aR）的相互作用促进草鱼的炎症和出血。为揭示草鱼感染GCRV后C5 a/C5 aR促炎反应的调控机制，采用免疫荧光法检测草鱼感染GCRV后肝脏和肌肉组织中C5的表达。采用RT-qPCR和western blot方法分析C5 a和C5 aR拮抗剂PMX 205在草鱼肾细胞系（CIK）中过表达对p38和ERK磷酸化以及IL-6和TNF-α基因和蛋白表达的影响。结果表明，草鱼感染GCRV后，肝脏和肌肉中C5 a蛋白的表达量呈先上升后下降的趋势，在感染后第5天达到高峰。在CIK细胞中，C5 a的过度表达显着增加C5 aR的表达、p38和ERK的磷酸化以及IL-6和TNF-α的过表达。PMX 205处理CIK细胞后，C5 a和C5 aR的表达、p38和ERK蛋白的磷酸化以及IL-6和TNF-α的表达均显著降低。这些结果表明，草鱼感染GCRV后，C5 a和C5 aR相互作用导致p38和ERK蛋白磷酸化，IL-6和TNF-α表达显著增加，最终诱导草鱼产生炎症反应。这些结果为通过调控C5 a和C5 aR的相互作用缓解草鱼出血病提供了理论依据。
Death of grass carp (Ctenopharyngodon idella) caused by grass carp reovirus (GCRV) infection severely restricts the development of the grass carp industry. The interaction of C5a and C5a receptor (C5aR) in the complement system promotes inflammation and aggravates bleeding in grass carps. To reveal the regulatory mechanism of the C5a/C5aR pro-inflammatory response after GCRV infection, the expression of C5 in the liver and muscle tissues of grass carp after GCRV infection was analyzed using immunofluorescence. The effects of over expression of C5a and the C5aR antagonist PMX205 in grass carp kidney cell line (CIK) on the phosphorylation of p38 and ERK and the gene and protein expression of IL-6 and TNF-α were also analyzed using RT-qPCR and western blot. The results showed that the expression of the C5a protein in the liver and muscle of grass carp infected with GCRV first increased and then decreased, and the C5a protein level peaked on the 5th day after infection. In CIK cells, over expression of C5a significantly increased C5aR expression, phosphorylation of p38 and ERK, and overexpression of IL-6 and TNF-α. Treatment of CIK cells with PMX205 significantly decreased the expression of C5a and C5aR, phosphorylation of p38 and ERK proteins, and expression of IL-6 and TNF-α after GCRV infection. These results implied that the interaction of C5a and C5aR in grass carp resulted in the phosphorylation of p38 and ERK proteins, a significant increase in the expression of IL-6 and TNF-α, and ultimately induction of an inflammatory response of grass carp infected with GCRV. These results provide a theoretical basis for relieving grass carp hemorrhagic diseases by regulating the interaction of C5a and C5aR.