Study On The Mechanism Of Resveratrol Regulating Fat Deposition Through Adiponectin Signaling Pathway In Pig

Resveratrol(RES)is a plant polyphenolic substance with various biological functions.The Adiponectin(adipoQ)signaling pathway plays an important role in fat metabolism.In order to explore the molecular mechanism of RES regulating pig fat deposition,we used qPCR,Werstern blotting,RNA-seq and other experimental methods to study the molecular mechanism of RES regulating fat deposition through adiponectin signaling pathway in pig organic and cellular levels.In this study,twenty-four Du Chang Da crossbreed pigs with about 65 kg body weight were randomly divided into 4 treatment groups:control group(feeding basic diet),200 mg/kg(basal diet + 200 mg/kg RES),400 mg/kg(basic diet + 400 mg/kg RES),600 mg/kg(basic diet + 600 mg/kg RES),each group had 6 pigs.These pigs fed for 41 days,weighing up to 110 kg,weighing the fasting weight of each pig,later for slaughter determination.The subcutaneous fat and liver tissue samples collected,used for RNA extraction and sectioning.We used qRT-PCR and Werstern blotting measured the expression of adiponectin signaling pathway related genes in tissue samples,studied on the molecular mechanism of fat deposition through adiponectin signaling pathway regulates in pigs.The research results are as follows:1.Compared with the control group,the thickness of the backfat was significantly decreased in the three experimental groups,and the corpus callosum rate was significantly reduced.HE staining of adipose tissue sections showed that the volume of fat cells decreased.2.Gene expression analysis in pig adipose tissue showed that genes up-regulated in the three experimental groups included adipoxin signaling pathway genes AdipoQ,AdipoRl,AdipoR2,AMPKa,PPARa,and mitochondrial production key genes SIRT1,PGC-1α,NRF1,lipolysis genes HSL,ATGL,fatty acid oxidation metabolism genes LPL,CPT1,and the ACC gene whose expression is down-regulated are the key genes of fat synthesis.The results showed that RES could regulate the expression of downstream fat metabolism genes,activate fat metabolism,inhibit fat synthesis,reduce body fat deposition by activating two signaling pathways AdiopQ-AdipoR1-AMPKα-PGC-1α and AdiopQ-AdipoR2-PPARa.3.RES(30 μM)treatment of 3T3-L1 cells could inhibit cell differentiation and reduce lipid droplet formation.RES could upregulate the genes expression of AdipoQ.AdipoRl,AdipoR2,AMPKa,PPARa,HSL,ATGL,LPL,CPT1 in cell.RES could regulate fatty acid metabolism through adiponectin signaling and reduce intracellular lipid droplet formation.4.The results of RNA-seq sequencing showed that 14777 genes were identified in the subcutaneous adipose tissue of control group and RES group(400mg/kg),including 499 lncRNAs.There were 82 differential genes in the two groups.including 2 up-regulation and 3 down-regulation of lncRNA;these differentially expressed genes are mainly involved in adipocytokine signaling pathway,Wnt signaling pathway,PI3K-Akt signaling pathway,and MAPK signaling pathway,which were associated with glycolipid metabolism.Functional predictions suggest that RES maight be up-regulated MSTRG9710 lncRNA acts on the adiponectin signaling pathway to regulate fat metabolism in pigs.5.Compared with the control group,the levels of adiponectin and high-density lipoprotein(HDL)in the serum of the three experimental groups were significantly increased,and the total cholesterol(TC)content was significantly decreased.Liver tissue sections HE and Oil red O staining showed no significant difference in liver tissue appearance in the four groups of pigs.In liver,the enzyme activity of GSH-Px,a key antioxidant enzyme,of the three experimental groups,was significantly higher than that of the control group.The content of lipid peroxidation metabolite MDA was significantly lower than that of the control group.RES might protect liveriver function.The results of gene expression analysis in liver tissue showed that the expression levels of AdipoR2,PPARa,CPT1,LXRa and ABCA1 were significantly increased in the three experimental groups compared with the control group.HepG2 cells were treated with RES(50 μM),the expression trends of related genes in cells were consistent with those in vivo.The results showed that RES promoted fatty acid oxidation and cholesterol utilization by activating the AdipoR2-PPARa signaling pathway,and enhanced the fat metabolism of the liver.In summary,RES enhances the expression of key genes of lipolysis and metabolism,inhibits the expression of key genes in fat synthesis,and promotes fat metabolism and inhibits fat synthesis and reduces subcutaneous fat by activating adiponectin-signaling pathway in pig subcutaneous fat and liver.RES can reduce the thickness of subcutaneous fat in pigs,enhance hepatic lipid metabolism,and reduce carcass fat percentage.