Effect Of Insulin, Glucagon, Insulin Like Growth Factor-â…  On IGF-â…  MRNA, PEPCK MRNA Expression Of In In Vitro Culture Bovine Hepatocytes

The objective of present study was to detect the effect of Insulin (INS) Glucagon(GN) on IGF-I (Insulin-like growth factor I )and all the three on on Phosphoenolpyruvate carboxykinase(PEPCK) mRNA expression of in vitro culture bovine hepatocytes through quantity RT-PCR method. Firstly, PEPCKcDNA, IGF-I cDNA, β-actincDNA genes were successly cloned, optimization PCR conditions were detected, their sequences were identical with Genebank reported. Secondly,Difference concentration Insulin, Glucagon , Insulin-like growth factor- I were added to in vitro culture bovine hepatocytes to observe the changed of IGF-I mRNA and PEPCKmRNA abundance. Concentration of the additives was as INS(0nmol/ml 1 nmol/ml 10 nmol/ml 100 nmol/ml 1000 nmol/ml), GN(0pg/ml 1 pg/ml 10pg/ml 100 pg/mk 500 pg/ml),IGF-I (Ong/ml 1ng/ml 10ng/ml 15ng/ml 20ng/ml 30ng/ml 40 ng/ml 50ng/ml 60ng/ml 100 ng/ml 150 ng /ml) After culture 12h, IGF- I mRNA, PEPCKmRNA expression was quantitatively determinated by competive PCR. The results are that IGF- I mRNA abundance in bovine hepatocytes enhanced significantly with increasing insulin concentrations in the media (P<0.01)and the high concentrations of insulin did not inhibit IGF-I mRNA abundance. On the contrary , glucagon inhibit IGF-I mRNA abundance, the experiment group is significantly difference to control group excep of low concentration(P<0.01). When concentrations of IGF- I were increased from Ong/ml to 150 ng/ml, PEPCK mRNA abundance in bovine hepatocytes decreased significantly . With increasing insulin, PEPCK mRNA abundance in bovine hepatocytes inhibit...