Regulating Metabolism Of Apolipoprotein B On In Vitro Model Of Fatty Liver

Apolipoprotein is a kind of polymorphism protein on molecular weight, immunity, metabolism, identification mark on receptor integrating, and participation in integration and metabolism on lipoprotein and its receptor of cell surface. It is important in lipoprotein metabolism. In this study, the lipid metabolism mechanism of Apolipoprotein was investigated by in vitro experiments.The rat primary cultured hepatocytes were isolated by two-step collagenase digestion method and treated with gradient concentration of DL-ethionine. After the hepatocytes were treated for 24~48h, the content of AST, ALT in the medium and TG, apoA, apoB, LDL and HDL in the hepatocytes were measured. The fatty droplets in hepatocytes could be observed by oil red O staining. The results as follows: (2~4)×108 cells per rat were obtained and the viability was about 95%. Hepatocytes grew well, had a good morphology, and could be used for investigation of lipid metabolism. Treated by gradient concentration of DL-ethionine with different time. The content of AST, ALT in treated hepatocytes increased gradually as the DL-ethionine concentration increased and time course. The content of TG increased at the same concentration, it's more significant in 24h than that in 48h. The content of apoAⅠincreased significantly in model group compared with control(P<0.01)in 24h, and there was no significant difference between 24h and 48h. The content of apoB decreased significantly in 2.0, 3.0, 4.0, 5.0 mmol/L model group compared with control(P<0.01)in 24h and 48h. The content of LDL, HDL increased significantly in 2.0, 3.0, 4.0, 5.0 mmol/L model group compared with control(P<0.01) in 24h and 48h. Watch under light microscope by oil red O staining, obvious accumulation of triacylglycerol in hepatocytes and gradient increase of red fatty droplets compared with control group, some became flakiness and membrane edge had no integrity, nucleus was round and blue-staining. These consistent with morphological characteristics of hepatocytes fatty degeneration.According to above experiment, rat primary cultured hepatocytes were treated with 2.0mmol/L DL-ethionine for 24h to made fatty live in vitro model. Treated with gradient concentration of Rhizoma Alismatis crude extract, the changes of TG, apoB and HDL were measured by above method and lipid metabolism mechanism of apolipoprotein were investigated. The results as follows: the content of TG, LDL decreased obviously and the content of apoB increased obviously in the groups which were treated with gradient concentration of Rhizoma Alismatis crude extract. The results in 24h were more significant than that in 48h. Under the gradient concentration of Rhizoma Alismatis crude extract, 5.0μg/mL was the best concentration.At first, ideal model was established to investigate preferably fatty liver. In this study, it succeeded to made fatty liver in vitro model by DL-ethionine induced hepatocytes adipose degeneration. Through observing dynamic succession, we can research directly cellular mechanism of fatty liver degeneration. From biochemical view, apolipoprotein regulated action on pathogenesis of fatty liver were investigated further by the technique.