Study On Two LncRNA And MiR-451 Of Grass Carp Regulating The Inflammatory Response Induced By Aeromonas Hydrophila

Grass carp(Ctenopharyngodon idella)is the fish with the highest aquaculture production in the world.As the scale of aquaculture expands and the density of aquaculture increases,it is easy to cause various diseases and affect the production of aquaculture.Aeromonas hydrophila infection can induce grass carp bacterial diseases.This paper conducts follow-up functional analysis on lncRNA-SUMO3,lncRNA-HDMO13 and miR-451,and finds that grass carp lncRNA-SUMO3 and lncRNA-HDMO13 can be combined with miR-21 and miR-142a-3p regulate the inflammatory response induced by A.hydrophila,miR-451 can regulate the inflammatory response induced by A.hydrophila by targeting hdac8 and npr2.The main research results of the paper are as follows:1.Based on the previous transcriptome of the research group,compared with four databases,the intersection obtained is lncRNA.According to the upstream and downstream 5K of the target genes,we conducted co-localization and co-expression screening.Finally,we selected two lncRNAs that may be involved in bacterial infections for research,namely lncRNA-SUMO3 and lncRNA-HDMO13.During the infection of grass carp by Aeromonas hydrophila,the expression patterns of lncRNA-SUMO3 and lncRNA-HDMO13 were increased first and then decreased.The dual-luciferase reporter assay system confirmed that lncRNA-SUMO3 and lncRNA-HDMO13 can interact with miR-21 and miR-142a-3p combination.Overexpression of lncRNA-SUMO3 and lncRNA-HDMO13 can significantly inhibit the expression of miR-21 and miR-142a-3p;after inhibiting lncRNA-SUMO3 and lncRNA-HDMO13,the expression levels of miR-21 and miR-142a-3p increase significantly.In overexpression or inhibition experiments of lncRNA-SUMO3 and lncRNA-HDMO13,the expression of downstream inflammatory factors tnf-α,il-12,il-6,il-1β,il-8 and tgf-β changed significantly,indicating that lncRNA-SUMO3 And lncRNA-HDMO13 can affect downstream inflammatory factors,tnf-α,il-12,il-6,il-1β,il-8 and tgf-β by combining miR-21 and miR-142a-3p.2.Based on the existing miRNA-m RNA targeting relationship of the research group,the expression patterns of miR-451,hdac8 and npr2 were detected at different time points after infecting grass carp CIK cells with Aeromonas hydrophila.It was found that the expression trend of miR-451 was opposite to that of hdac8 and npr2.RNAhybrid software predicted that miR-451 has hypothetical binding sites with hdac8 and npr2.The dual-luciferase reporter system showed that miR-451 could bind to the3’UTR binding sites of hdac8 and npr2 to inhibit its expression,indicating that hdac8 and npr2 are the target genes of miR-451.Overexpression of miR-451 could significantly inhibit hdac8 and npr2;inhibition of miR-451 could significantly increase the expression of hdac8 and npr2.Furthermore in overexpression or inhibition experiments of miR-451,the expression of downstream proinflammatory factors il-6,il-8,il-12 and il-1β changed significantly,indicating that miR-451 can target hdac8 and npr2 to regulate downstream proinflammatory factors,il-6,il-8,il-12 and il-1β.