Discovery Of Immune-related Genes And Proteins And Analysis Of Integrin Gene Family In Liver Tissue Of Tongue Sole Infected With Vibrio Eel

Half-smooth tongue sole(Cynoglossus semilaevis)is susceptible to vibrio infection in large-scale culture,among which the common Vibrio anguillarum infection will cause a high mortality rate and huge economic loss.Currently,there is a lack of studies on transcriptomics and proteomics combination to analyze the immune mechanism of V.anguillarum infection in C.semilaevis.The liver is a multi-functional organ with metabolism,substance synthesis,detoxification,and various immune functions,and its role in immunity has attracted more and more attention.In this study,transcriptomic and proteomic techniques were used to analyze the changes of immune genes and proteins in the liver of C.semilaevis infected with V.anguillarum infection.The metabolic pathways involved in differential genes and proteins were analyzed.In addition,the integrin gene family of C.semilaevis was analyzed,which provided experimental data for the study of immune mechanism of C.semilaevis against V.anguillarum infection and the role of integrins in innate immunity of fish.The main findings were as follows:1.A total of 1306 differentially expressed genes were identified in the liver tissues after 24 hours infection with V.anguillarum.Among them,345 differentially expressed genes were found in the pairwise comparison of obvious symptoms of infection(HOSG)and PBS control group(CG),and 961 differentially expressed genes were found in the pairwise comparison of no obvious symptoms of infection(NOSG)and CG,and 164 differentially expressed genes were found in the pairwise comparison of NOSG and HOSG.Some genes closely related to pattern recognition receptors,complement synthesis,the activation of autophagy,inflammatory response and lipid synthesis,such as TLR5 complement C3,C5,Sgh C1 q,LC3,IL-1β,IL1R2,HSP30,HSP90 and PPARγ were up-regulated,TLR3 and TLR7 were down-regulated.Analysis of KEGG enrichment pathways showed significant immune responses including toll-like receptors(TLRs),complement and coagulation cascades,NOD-like receptors(NLRs),MAPK and phagosome,which suggested that these five signaling pathways played an important role in the immune response of the liver of C.semilaevis.2.A total of 497 differentially expressed proteins(DEPs)were identified in liver tissues 24 h after infection with V.anguillarum.Among them,143 DEPs were found in HOSG and CG pairwise comparison,218 DEPs were found in NOSG and CG pairwise comparison,and 136 DEPs were found in NOSG and HOSG pairwise comparison.Some proteins closely related toapoprotein synthesis,integrin and phagocytosis,such as APOE,ITGβ1 and MRC1 were up-regulated.Analysis of KEGG enrichment pathways showed significant immune responses including phagosome,ECM-receptor interaction,MAPK and PPAR signaling pathway.3.Combining transcriptomics and proteomics analysis showed that the phagosome,MAPK and ECM-receptor interaction signaling pathway and ITGβ1,C3,C5,MRC1,APOE,PPARγ were up-regulated at both m RNA and protein levels,and the protein-protein interaction network analysis showed that TP53I3 and THBS1 were at the pivotal position,suggesting that they played an important role in the immune strategy of recognizing and resisting the invasion of V.anguillarum.4.q RT-PCR analysis showed that the differentially expressed FN1,MRC1,APOE and THBS1 genes were relatively high in the liver tissue after V.anguillarum infection,suggesting that these genes played an important role in the innate immune response of liver.5.The integrin gene family of C.semilaevis were identified and analyzed.A total of27 members were identified,the motif composition and distribution of ITGα10 and ITGα11 were highly similar,suggesting that these two proteins had similar functions,and that ITGβ1 may be functionally differentiated.ITGα4 and ITGα5 were evolutionarily distinct from other α subunits.The integrin α-subunit and β-subunit of C.semilaevis were significantly different.The composition and distribution of β-subunit protein were more similar than that of α-subunit protein,and the length of INB domain was more uniform.q PT-PCR analysis showed that members of the integrin family were widely expressed in liver,intestine,gill,spleen,head kidney and blood tissues.The spatiotemporal expression patterns of them in the liver,intestines,gills and spleen after V.anguillarum infection showed different degrees of differential expression,which were involved in the innate immune response of C.semilaevis to pathogen stimulation.This study was the first to combine transcriptomic and proteomic data to investigate the immune response of the liver of C.semilaevis to V.anguillarum infection,and a number of key immune genes and proteins were obtained.The identification and analysis of integrin in C.semilaevis will provide some basis for the study of immune function in the future.This study laid a foundation for further revealing the immunologic mechanisms of immune-related genes and proteins in the process of disease resistance of C.semilaevis,and also provided some theoretical references for rational disease prevention and treatment and pathogenesis exploration of C.semilaevis.