Hormonal Regulation Of Stearoyl Coenzyme-A Desaturase Gene Expression In Primary Cultures Of Newborn Calf Hepatocytes

The express of SCD mRNA was effected by insulin, glucagon and NPY in the hepatocytesof new born calf. Healthy new born calf were chosen and the hepatocytes were dissociated withperfusion succesefully, The ability of synthetized urea, the ability of secreted albumen and lactic aciddehydrogenase spillage were detected in the culture solution. The damage and recovery of hepatocyteswas detected. The amplication rate ofβ-actin and SCD was detected by fluorescence PCR withSRBR greenⅠand plasmid. The hepatocytes were treated with insulin (0,5,10,20,50IU/m),glucagons(0,50,100,500,1000pg/ml) and NPY (0,50,100,500,1000pg/ml) after cultured 72h.ThemRNA was extractd and correspondencet change of SCD mRNA was detected by fluorescence PCRaccording to change ofβ-actin. The results indicate that:1.The perfusion can dissociatehepatocytes as well as can,the survival rate of hepatocytes maintain up to 90%. The ability ofsynthetized urea, the ability of secreted albumen begin to descend after 3 d, lactic acid dehydrogenase islower after 3d.That show the damage of hepatocytes are recoverd after 3d. 2. Twoequations(Y=34.14-2.95*X (Ⅰ) and Y=35.46-2.97*X (Ⅱ)) were getten according to plasmidconcentration log and CT. The two equations show that SCD andβ-actin have same amplicationrate. Relative amplication of SCDmRNA can be computed using 2^{-ΔΔCt} method. 3 The SCD mRNAexpresss of hepatocytes is dose-dependently increased by insulin. There is significant difference amongall group.4 The SCD mRNA expresss of hepatocytes is dose-dependently depressed by glucagons Thereis extremely significant difference among all group.5 The SCD mRNA expresss of hepatocytes isincreased by NPY There is significant difference between all group except treated with 50pg/ml and500pg/ml.6 Insulin and NPY increase SCDmRNA express and increase liver fat storing.Glucagondepress SCDmRNA express and inh inhibite liver fat storing.