| Abstrakt: |
The first branch point in gluconeogenesis occurs at the conversion of pyruvate to oxaloacetate. To determine the amount of lactate carbon reaching glucose via the direct pyruvate carboxylase pathway versus the tricarboxylic acid cycle, adult rat hepatocytes in primary culture were incubated for 2 h with one of the following isotopic substrates: [1-14C]lactate, [U-14C]lactate, or [1,2-14C]acetate. Production of 14CO2 and [14C]glucose from each substrate was assessed. The amount of lactate carbon 2 and 3 incorporated into glucose or oxidized to CO2 was determined by subtracting values using [1-14C]lactate from those using [U-14C]lactate. After quantitation of CO2 formed from carbons 2 and 3 of lactate, the amount of these carbons incorporated into glucose via the tricarboxylic acid cycle can be determined by simple proportionality from the ratio of label incorporated into glucose or CO2 from [1,2-14C]acetate. The remaining carbons 2 and 3 of lactate incorporated into glucose are derived from the pyruvate carboxylase pathway directly. Ethanol which on oxidation provides NADH and acetate decreased lactate oxidation and enhanced the pyruvate carboxylase pathway. Glucagon increased carbon flux through both pathways but primarily through the pyruvate carboxylase pathway. In summary, a simple model is presented to examine carbon flux from lactate via the pyruvate carboxylase and tricarboxylic acid pathways during gluconeogenesis. |
