Analysis Of Blood Index And Screening Of Differential MiRNA In Circulating Exosomes Of Obese Canine

In recent years,the obesity of dogs has appeared frequently.Obesity not only affects the canine appearance,but also shortens the canine lifespan,affects animal welfare,and poses a great threat to the dog’s health.This experiment provides basic data for finding new biomarkers of canine obesity by analyzing blood indexes of obesity canines and screening differential miRNAs in circulating exosomes.Select 6 pet canines and record basic information.Canines were divided into obesity group and control group by using BCS.Blood was collected through the veins of the forelimbs and blood samples were taken.Detect the blood routine indicators,and then detect the concentration of serum biochemical indexes TC,TG,LDL-C and other indexes;ELISA method to detect the concentration of serum IL-6,TNF-α,ADPN and other indexes.Plasma exosomes were extracted by ultra-high-speed centrifugation,the external morphological characteristics of exosomes were identified by transmission electron microscopy,the particle size distribution of exosomes was identified by nanoparticle tracking analyzer,and the protein of surface exosomes was identified by Western blotting.High-throughput sequencing for exosome miRNA,screening the differential miRNA of plasma exosomes in the control and obesity canines,and performing GO function annotation and KEGG pathway annotation on the differential miRNA.The results show:1.Blood index analysis: Compared with the control group,the number and proportion of mononuclear cells in the obese group increased,and the serum TC,TG,LDL-C,GLU,ALT,AST,and ALP indicators increased.The concentrations of IL-6,TNF-α,LEP,INS,and LEP:ADPN increased significantly(P<0.01),and the concentration of ADPN decreased significantly(P<0.01).In the correlation analysis,BCS was significantly proportional to the concentration of IL-6 and TG(P<0.05),inversely proportional to the concentration of ADPN(P<0.05),and directly proportional to the concentration of LEP,INS,LEP: ADPN(P <0.01).AST concentration was significantly proportional to INS:GLU(P<0.05).2.High-throughput sequencing results: Target genes regulated by differentially expressed miRNAs in obesity canines are significantly enriched in transport catabolism,signal transduction,signal molecule interaction,endocrine and metabolic diseases,lipid metabolism,carbohydrate metabolism,amino acid metabolism,etc.15 miRNAs were significantly up-regulated and 31 miRNAs were significantly down-regulated.In summary,the blood routine indicators and kidney function indicators of obese canines are normal.The liver function indicators are high,the risk of liver disease is increased.The lipid metabolism indicators is increased,the cytokine indicators is significantly increased and the secretion of lipid metabolism-related hormones is significantly changed which indicated that there are disorder of lipid metabolism and low degree of inflammatory changes in obese canine,there is no inflammatory response.BCS is significantly associated with inflammatory cytokines and lipid metabolism-related hormones,indicating that inflammatory cytokines and lipid metabolism-related hormones are significantly related to the process of obesity.The pathways involved in the regulation of exosome miRNA are consistent with the symptoms of endocrine and metabolic disorders,abnormal digestive function,insulin resistance,and depression in the clinical symptoms of obese canines which indicated that the physiological and pathological changes caused by obesity will be reflected from the exosome differential miRNA.Therefore,miRNAs are differentially expressed in circulating exosomes which have exhibited huge potential in becoming new type of obesity-related biomarkers.