The Effect Of Rhizoma Alismatis On CytocromeP450of Dairy Cows Steatosis Hepatocytes

Fatty liver was the common disease on dairy cows. Generally, it happened on thehigh-yielding of perinatal dairy cows. Fatty liver could lead to decline on the milkproduction and accompany with other diseases, which brought serious economic loss to thedairy cow industry. In recent years, the incidence of fatty liver in dairy cows had anincreasing trend, but the occurrence mechanism of the fatty liver was still not very clear.Cytochrome P450enzymes were the main oxidases metabolic enzymes in the livermicrosome and played the very important role in the liver metabolism.The primary cultured hepatocytes were isolated by two-step collagenase perfusion.DL-ethionine was added into the hepatocytes to induce hepatocytes steatosis. The activitiesof alanine aminotransferase (ALT), aspartate aminotransferas (AST), superoxidedismutase(SOD) and the contents of triglycerides(TG), total cholesterol(TC), free fattyacid(FFA), malondialdehyde(MDA) were detected. The hepatocytes were stained by OilRed O method, and the changes of the hepatocytes morphology and the intracellular lipiddroplets were observed under microscope.According to above biochemical indices and the hepatocytes feature, the steatosishepatocytes model was established. The hepatocytes were treated with gradientconcentration of Rhizoma Alismatis for24h-48h. The activities of ALT, AST, SOD, andthe content of TG, TC, MDA, FFA were detected. The content of TNF-alpha was detectedby double antibody sandwich ELISA method. The hepatocytes morphology were observedby Oil red O. CYP450expression in the hepatocytes was analyzed by Western blot. Theeffect of CYP450in the formation of fatty liver was studied and the formation mechanismof fatty liver in dairy cow was explored to provide basis for treatment of the fatty liver indairy cows.The results showed, the hepatocytes of dairy cows grew well and had a goodmorphology, and they could be used for investigation of lipid metabolism. After thehepatocytes were induced by DL-ethionine, the activities of ALT, AST increased, theactivity of SOD decreased, the contents of TG, TC, FFA, MDA increased. The number ofthe lipid droplets increased and the volume of the lipid droplets became larger. The bestcondition to induce the hepatocytes steatosis was the treatment of2mmol/L DL-ethionine for24hours. After the hepatocytes were treated by Rhizoma Alismatis, the activities ofALT, AST decreased, SOD activity increased, the contents of TG, TC, FFA, MDA andTNF-alpha in the hepatocytes of dairy cows decreased. The number of the lipid dropletsdecreased and the volume of the lipid droplets became smaller. With the treatment ofgradient concentration of Rhizoma Alismatis, the expression of CYP450decreased bywestern blot methodThe steatosis hepatocytes model could be established successfully by2mmol/LDL-ethionine for24hours.10μg/mL Rhizoma Alismatis had protective effect on thesteatosis hepatocytes induced by DL-ethionine.