【摘要】 在鲤(Cyprinus carpio)感染鲤疱疹病毒Ⅲ型(CyHV-3)不同时期,通过分析鲤疱疹病毒标志基因TK和ORF72与自噬标志基因Beclin-1、LC3、P62的差异表达情况,探讨在CyHV-3感染中自噬的发生情况。研究表明:(1)抗病选育组与未选育组鲤TK和ORF72基因以及自噬标志因子Beclin-1、LC3、P62基因相对表达量均呈现先升高后降低的趋势; TK和ORF72基因相对表达量选育组均极显著低于未选育组(P<0.01)且在感染144 h达到最高;(2)选育组鲤Beclin-1、LC3、P62基因相对表达量在感染48h达到最高,且Beclin-1、LC3基因表达量极显著高于未选育组(P<0.001),P62显著高于未选育组(P<0.05);(3)未选育组鲤Beclin-1、LC3、P62基因相对表达量在感染96h达到最高且极显著高于选育组(P<0.001),而P62基因相对表达量持续在感染144h、192h极显著高于选育组(P<0.01),说明P62基因表达发生了聚集;(4) LC3蛋白表达趋势与基因相对表达量趋势一致。以上结果表明CyHV-3感染能够诱导鲤发生自噬,推测自噬体降解过程可能被抑制产生不完全自噬从而促进病毒复制感染。更多还原

【Abstract】 CyHV-3 infection causes a highly lethal disease in common carp, and it can be mainly detected by using TK and ORF72 gene markers. Autophagy is a stress protection mechanism in cells, mainly marked by Beclin-1,LC3, and P62 genes. This paper aimed to analyze the differential expression of CyHV-3 marker genes, TK and ORF72, in common carp(Cyprinus carpio) and autophagy marker genes Beclin-1, LC3, and P62 during different stages of CyHV-3 infection, and to explore the occurrence of autophagy in CyHV-3 infected common carp. The results showed that:(1) The relative expression of TK and ORF72 genes and the autophagy marker genes, Beclin-1,LC3, and P62, in the breeding group and the non-breeding group increased initially and then decreased. The relative expression of TK and ORF72 genes in the breeding group was significantly lower than that in the non-breeding group(P<0.01), and reached the highest at 144 h post-infection;(2) The relative expression of Beclin-1, LC3, and P62 genes in the breeding group reached the highest level at 48 h post-infection, and the expression of Beclin-1 and LC3 genes in the breeding group was extremely significantly higher than that in the non-breeding group(P<0.001), and P62 expression was extremely significantly higher in the breeding group than that in the non-breeding group(P<0.05);(3) The relative expression of Beclin-1, LC3, and P62 genes in the non-breeding group reached the highest level at 96 h post-infection and was significantly higher than that in the breeding group(P<0.001), while the relative expression of P62 gene was significantly higher at 144 h and 192 h post-infection in the non-breeding group(P<0.01), indicating that the expression of P62 gene was clustered;(4)The trend of LC3 protein expression was consistent with the trend of relative gene expression. The above studies indicate that CyHV-3 infection can induce autophagy in carp. It is speculated that the autophagy degradation process may be inhibited resulting in incomplete autophagy, thus promoting viral replication and infection. The probability of occurrence of complete autophagy needs further investigation. This study provides an important basis for the investigation of the mechanism of carp anti-CyHV-3 and the molecular mechanism of fish autophagy during viral infection.更多还原