Impact of flow rate on lactate uptake and gluconeogenesis in glucagon-stimulated perfused livers.

The impact of reduced hepatic flow on lactate uptake and gluconeogenesis was examined in isolated glucagon-stimulated perfused livers from 24-h-fasted rats. After surgical isolation, livers were perfused (single pass) for 30 min with Krebs-Henseleit (KH) bicarbonate buffer, fresh bovine erythrocytes (hematocrit approximately 20%), and no added substrate. After this "washout" period, steady-state perfusions were initiated with a second reservoir containing the KH buffer, bovine erythrocytes, [U-(14)C]lactate (10,000 dpm/ml), lactate (2.5 mM), and glucagon (250 microg/ml). Perfusion flow rate was adjusted to one of five rates (i.e., 1.8, 2.7, 3.9, 7.4, and 11.0 ml.min(-1).100 g body wt(-1)). After the perfusion, the liver was dissected out and weighed so as to establish the actual flow rate per gram of liver. The resulting flow rates ranged from 0.52 to 4.03 ml.min(-1).g liver(-1). As a function of flow rate, lactate uptake rose in a hyperbolic fashion to an apparent plateau of 2.34 micromol.min(-1).g liver(-1). Fractional extraction (FX) of lactate from the perfusate demonstrated an exponential decline with increased flow rates (r=0.97). At flow rates above 1.0 ml.min(-1).g liver(-1), adjustments in FX compensated for changes in lactate delivery, resulting in steady rates of lactate uptake and gluconeogenesis. Below 1.0.min(-1).g liver(-1) the increased FX was unable to compensate for the decline in lactate delivery and lactate uptake declined rapidly. Gluconeogenesis demonstrated similar kinetics to lactate uptake, reflecting its dominant role among pathways for lactate removal under the current conditions.

Lactate delivery (not oxygen) limits hepatic gluconeogenesis when blood flow is reduced.

The purpose of this study was to determine, using the isolated liver perfusion technique, whether the limiting factor for hepatic gluconeogenesis (GNG) from lactate was precursor delivery or oxygen availability during reduced flow rates of 0.85 or 0.60 ml.min(-1).g liver(-1). After a 24-h fast, three different experimental protocols were employed. Protocol 1 examined the impact on GNG when reservoir lactate concentration was maintained but oxygen delivery was elevated via increases in hematocrit (Hct). Elevating the Hct from 22.5+/- 0.8% to 30.9+/- 0.4% at a blood flow of 0.89+/- 0.01 ml.min(-1).g liver(-1) increased the oxygen consumption (Vo(2)) but did not augment GNG. Similarly, when the Hct was elevated from 22.5+/- 0.8% to 41.5+/- 0.7% at 0.59+/- 0.04 ml.min(-1).g liver(-1), Vo(2) was increased, but GNG was unaffected. Protocol 2 examined the impact on GNG when Hct was maintained but precursor delivery was elevated via increases in reservoir lactate concentration ([LA]). Specifically, elevating the [LA] from 2.31+/- 0.07 to 3.61+/- 0.33 mM at a flow rate of 0.82+/- 0.04 ml.min(-1).g liver(-1) significantly increased GNG. Similarly, elevating the [LA] from 2.31+/- 0.07 to 4.24+/- 0.37 mM at a flow rate of 0.58+/- 0.02 ml.min(-1).g liver(-1) increased GNG. Finally, we examined the impact of increasing both the oxygen and lactate delivery (Protocol 3). Again, Vo(2) was elevated with increased oxygen delivery, but GNG was not augmented beyond that observed with elevations in lactate delivery alone, i.e., Protocol 2. The results indicate that, during decrements in blood flow, GNG is limited primarily by precursor delivery, not oxygen availability.