The Effects Of Probucol On Mitochondria Of Experimental NAFLD Of Rats

Objective:In recent years,the incidence rate of non-alcoholic fatty liver disease (NAFLD)is increasing gradually,the onset of NAFLD has close correlation of 2 diabetes,hyperlipoidemia and insulin resistance.Though the pathogenesis of NAFLD is not yet completely clear,.Many studies have suggested that function disorder of mitochondria and oxidative stress play an important role in progression of NAFLD. Probucol is one of the most effective anti-oxidant.This study was designed to observe function disorder of mitochondria of NAFLD,the preventive effect of probucol on NAFLD and elucidate the mechanism,and then validate the important role of OS in the progression of NAFLD.Methods:Twenty four Sprague-Dawley rats weighing 210±10g were randomly divided into three groups:8 for normal control group(NC group)fed with regular chow,8 for model group(HF group)fed with high lipid chow and the others for probucol-intervention group(PB group)fed with high lipid chow,too.PB group rats were administered with PB at dose of 500mg/(kg.d)by intragastic infusion,the others were given considerable volume of CMC.All of the rats were killed at the end of eighteenth week.The whole liver,calculate the liver index(liver wet weight/body weight×100%).The seral ALT,AST,TG,TC,and the liver homogenate TG,TC,FFA were assayed.The lipoperoxidation makers and anti-oxidant substance in the serum and liver mitochondria,such as MDA,SOD,GSH-PX,mitochondria membrane potential and ATP enzyme,were assayed.The ratio of serous SOD to MDA was calculated,too.The liver histopathologic sections of HE staining were observed,and the ultramicrostructure of liver cell was observed with transmission electron microscope too.Expression of UCP2 in the liver were detected by immunohistochemistry and RT-PCR.UCP2mRNA and PPARγmRNA were detected RT-PCR.The statistical analysis of the data was done with the statistical package of SPSS 11.0.Result:By the end of the eighteenth week,The liver cell of structure of NC group is approximately normal.In HF group,there is severe hepatic steatosis and mild to moderate inflammatory infiltration and point flake or spotty necrosis and fibrosis in the liver.TEM observation showed that part or most of mitochondrial ridge fuse with membrane or disappeared.Probucol significantly improve both pathology and ultrastructure of the liver.Compared with the NC group,the liver wet weight,liver index, the serous TC and LDL-c,the liver homogenate FFA concentration,of HF group were significantly increased(P＜0.05),and HDL-c is significantly decreased(P＜0.05).The administration of probucol decreased the serous TC,LDL-c and the liver homogenate FFA concentration by 33.1%,21.5%and 13.9%(P＜0.05).Compared to NC group,the MDA concentration in both serum and liver mitochondria of the HF group are significantly increased(P＜0.05),liver mitochondria SOD,GSH-PX and serous SOD/MDA are significantly decreased(P＜0.05).While,PB administration signifycantly decreased both serum and liver mitochondria MDA,and increased liver mitochondria SOD,GSH-PX and serous SOD/MDA(P＜0.05).As to the HF group,both serum ALT and AST are significantly increased,serous ALT and AST of PB group significantly decreased(P＜0.05).As to the NC group,liver mitochondria Na-K ATP enzyme,Ca-Mg ATP enzyme of the HF group decrease(P＜0.05),liver mitochondria Na-K ATP enzyme,Ca-Mg ATP enzyme of PB group increased significantly as to HF group(P＜0.05),but decreased as to NC group.Protein and mRNA of UCP2 were expressed un-regulated in liver of HF group,PB group are significantly decreased (P＜0.05).PPARγmRNA were expressed un-regulated in liver of HF group,.PB group are significantly decreased(P＜0.05)Conclusion:We successfully prepared rat model of NAFLD.There is function disorder of mitochondria,both liver local and systemic oxidative injury of the NAFLD rats.Probucol can reduce the degree of liver steatosis,that might be partially dependent on the antioxidant capacity of liver,Probucol prevents liver from oxidative injury,and thus delays non-alcoholic steatohepatitis progress.OS is an important pathophysiologicals mechanism to promote the progress of NAFLD,and antioxidant therapy is a potential treatment for NAFLD.