MiR-130c-5p靶向乌鳢水泡病毒g基因抑制病毒增殖

MicroRNA(miRNA)是一类内源性的非编码RNA，参与调节病毒复制、细胞增殖或凋亡以及肿瘤发生等生物反应。本研究探讨了miR-130c-5p在乌鳢水泡病毒(snakehead vesiculovirus, SHVV)感染中潜在靶基因g的靶向关系以及对病毒复制的影响。本研究以斑点叉尾鮰卵巢(Channel catfish ovary, CCO)为实验材料，通过qRT-PCR和Western blot测定SHVV不同感染时间和感染剂量条件下，病毒基因水平和蛋白水平以及miR-130c-5p变化情况。此外，将SHVV的g基因上miR-130c-5p对应的靶序列克隆到质粒pmirGLO，构建质粒pmirGLO-G用于双荧光素酶报告实验进行靶基因验证。结果显示，随着SHVV感染时间及剂量的不断增加，miR-130c-5p和g基因的表达水平都显著上调。进一步实验证明，miR-130c-5p类似物和pmirGLO-G质粒共转染可显著抑制荧光素酶活性强度，而转染miR-130c-5p抑制剂则明显上调了pmirGLO-G报告载体的荧光信号。此外，miR-130c-5p的过表达显著降低了病毒G基因的mRNA及蛋白表达，而抑制miR-130c-5p的表达则上调了g基因的mRNA及蛋白的表达水平。研究结果表明，miR-130c-5p通过靶向SHVV的g基因，引起G蛋白的降解，从而抑制SHVV的增殖。本研究结果为理解microRNA调控SHVV的致病机制提供了重要基础，为抗SHVV疫苗等药物的研发提供了理论支持。

MiR-130c-5p targets the snakehead vesicular virus g gene to inhibit viral proliferation

MicroRNAs (miRNAs) are a class of endogenous non-coding RNAs involved in regulating biological responses such as viral replication, cell proliferation or apoptosis, and tumorigenesis. In this study, we investigated the targeting relationship of miR-130c-5p in the potential target gene g in snakehead vesicular virus (SHVV) infection and its effect on viral replication. The viral gene level and protein level as well as miR-130c-5p changes were determined by qRT-PCR and Western blot under different conditions of infection time and infection dose of SHVV, using channel catfish ovary (CCO) as the experimental material. In addition, the target sequence corresponding to miR-130c-5p on the g gene of SHVV was cloned into the plasmid pmirGLO, and the plasmid pmirGLO-G was constructed for target gene validation by dual luciferase reporter assay. The results showed that the expression levels of both miR-130c-5p and g genes were significantly up-regulated with increasing SHVV infection time and dose. Further experiments demonstrated that co-transfection of the miR-130c-5p analogue and pmirGLO-G plasmid significantly inhibited the intensity of luciferase activity, whereas transfection of the miR-130c-5p inhibitor significantly up-regulated the fluorescence signal of the pmirGLO-G reporter vector. In addition, overexpression of miR-130c-5p significantly reduced the mRNA and protein expression of the viral G gene, whereas inhibition of miR-130c-5p upregulated the mRNA and protein expression levels of the g gene. The results suggest that miR-130c-5p inhibits SHVV proliferation by targeting the g gene of SHVV and causing degradation of the G protein. The results of this study provide an important basis for understanding the microRNA regulation of the pathogenic mechanism of SHVV and provide theoretical support for the development of an anti-SHVV vaccine and other drugs.