Effect Of Propionate, Pyruvate And β-Hydroxybutyric Acid On PC MRNA And PEPCK MRNA Expression Of In Vitro Culture Bovine Hepatocytes

The objective of present studies was to detect the effect of propionate, pyruvateand β-hydroxybutyric acid on pyruvate carboxylase (PC)mRNA and PEPCKmRNAexpression of in vitro culture bovine hepatocytes through quantity RT-PCR method.Firstly, Compared to PCmRNA, PCDNA has an intron sequence. Competive DNAtemplate of pyruvate carboxylase (PC) cDNA was constructed through the methods ofintron sequence and insertion sequence. Results displayed that PCcDNA and PCDNAgenes were successly cloned, their sequences were identical to 99.5% and 99.6% ofthat with Genebank report, so we construct the competitive DNA templat. Secondly,Difference concentration propionate, pyruvate and β-hydroxybutyric acid were addedto in vitro culture bovine hepatocytes to observe the changed of PCmRNA andPEPCKmRNA abundance. Concentration of the additives was as propionate, 0mmol/L,1.5mmol/L, 2.5mmol/L, 3.5mmol/L, 4.5mmol/L, 8.5mmol/L,11.5mmol/L(no thisconcentration of PEPCKmRNA); pyruvate, 0mmol/L, 1.0mmol/L, 2.0mmol/L(nothis concentration of PCmRNA), 4.0mmol/L, 8.0mmol/L , 12.0mmol/L; β-Hydroxybutyric acid, 0mmol/L, 0.5mmol/L, 0.75mmol/L, 1.0mmol/L,1.5mmol/L,3.0mmol/L. after culture 24h, PCmRNA and PEPCKmRNA expressionwas quantitatively determinated by competive PCR. The results showed thatPEPCKmRNA and PC mRNA abundance in bovine hepatocytes enhanced significantlywith increasing propionate concentrations in the media (P<0.01) and the highconcentrations of propionate did not inhibit PC mRNA abundance. There was asignificant positive correlation between propionate concentrations in the media and PCmRNA abundance(r=0.94, p<0.01). The different concentrations of pyruvate from 0mmol/L to 12mmol/L were added to the media of in vitro culture bovine hepatocytes.PC mRNA and PEPCK mRNA abundance in bovine hepatocytes enhanced...