传染性脾肾坏死病毒感染促进鳜脑组织细胞脂肪酸合成及β-氧化OA

We used transcriptomics,metabolomics,inhibitor experiments,reverse transcription quantitative PCR(RT-qPCR),quantitative PCR(qPCR),and western blotting(WB)to investigate the regulatory effects of infectious spleen and kidney necrosis virus(ISKNV)infection on fatty acid metabolism in Chinese perch brain(CPB)cells by exploring the relationship between ISKNV replication and host cell lipid metabolism.Multi-omics integrative analysis revealed a significant increase in the transcriptional expression of fatty acid synthesis-related genes(ACC1,FASN)and fatty acid oxidation-related genes(ATGL,HSL,and CPT)at 24 and 72 hours post-infection(hpi)with ISKNV.The content of various fatty acid metabolites increased at 24 hpi but decreased significantly at 72 hpi.Further experiments confirmed that at 36 hpi of ISKNV infection,the mRNA and protein levels of key fatty acid synthesis proteins(ACLY,ACC1,and FASN)were significantly upregulated;the mRNA and protein levels of key fatty acid oxidation proteins(ATGL and CPT1)and rate of fatty acid oxidation were significantly increased at 72 hpi.Inhibition of key fatty acid synthesis enzymes ACC1 and FASN using TOFA and C75 significantly suppressed the transcription and protein expression of ISKNV MCP and copy number in the extracellular supernatant at 72 hpi.Inhibition of key fatty acid oxidation enzyme CPT1 with Etomoxir also significantly reduced MCP protein expression and extracellular supernatant copy number at 72 hpi.These results indicate that ISKNV infection activates fatty acid synthesis and oxidation in CPB cells to support its own replication,providing a theoretical basis for an in-depth understanding of the pathogenic mechanism of ISKNV and antiviral strategies.