Resveratrol Activated PGC-1α To Relieve NEFA-induced Hepatocyte Injury In Dairy Cows

This study aimed to investigate the protective effect of resveratrol(RES)on bovine hepatocytes damage induced by non-esterified fatty acid(NEFA).Blood and liver tissue samples of healthy and ketotic cows were collected to determine glucolipid metabolism,oxidative stress and inflammatory factors,and it was found that cows with ketosis showed abnormal glucose and lipid metabolism,oxidative stress and inflammatory response.The rise of NEFA concentration is the main cause of this series of abnormal reactions.Subsequently,NEFA-induced bovine hepatocytes were used to establish hepatocyte injury models in vitro,and the protective effect of RES on NEFA-induced bovine hepatocytes was verified by adding RES.Cell viability was detected by CCK-8.The mRNA expression of mitochondrial transcription factor A(Tfam),peroxisome proliferator-activated receptor γ coactivator 1-α(PGC-1α),superoxide dismutase(SOD),glutathione peroxidase(GSHPX),malondialdehyde(MDA),nod-like receptor protein 3(NLRP3),interleukin-6(IL-6),interleukin-8(IL-8),cysteinyl aspartate specific proteinase-3 and cysteinyl aspartate specific proteinase-3 were detected by qRT-PCR,and the changes of mitochondrial membrane potential(MMP)of bovine hepatocytes were detected by JC-I probe.Reactive oxygen species(ROS)production was detected by ROS kits.The results were as follows:1)in vivo studies have showed that compared with healthy cows,an abnormal glucolipid metabolism,oxidative stress,inflammatory response and liver injury were happened in ketotic cows.2)CCK-8 showed that cell viability was decreased by high concentration of NEFA,which was reversed by the addition of RES.3)qRT-PCR results showed that the mRNA expression of B-cell lymphoma-2(Bcl-2),Tfam,PGC1α,SOD and GSH-PX were significantly increased after RES addition(P<0.05),while the mRNA expression of MDA,caspase-3,caspase-9,NLRP3,IL-6 and IL-8 were significantly decreased(P<0.05).4)the MMP of bovine hepatocytes detected by JC-1 probe showed that the addition of RES alleviated the mitochondrial damage induced by NEFA.5)The result of ROS assay showed RES could reduce NEFA-induced oxidative stress in bovine hepatocytes.6)After interfering PGC1α,RES could not relieve NEFA-induced mitochondrial damage,oxidative stress,inflammatory response and inhibit apoptosis of bovine hepatocytes.In conclusion,this study confirmed that exogenous RES alleviated NEFA-induced mitochondrial damage,oxidative stress,inflammatory response and inhibited apoptosis of dairy liver cells by activating PGC-1α.The addition of RES can provide theoretical basis for the prevention of important reactions such as mitochondrial damage,oxidative stress,inflammatory response and apoptosis during the development of ketosis in dairy cows and the development of RES in practical production.