Spolarics Z, Lang C H, Bagby G J, Spitzer J J
Department of Physiology, Louisiana State University Medical Center, New Orleans 70112.
Am J Physiol. 1991 Aug;261(2 Pt 1):G185-90. doi: 10.1152/ajpgi.1991.261.2.G185.
This study assessed and compared the rate of glucose utilization, activity of the hexose-monophosphate shunt (HMS), and the oxidation of glutamine, lactate, and palmitate in Kupffer (KC), endothelial (EC), and parenchymal liver cells (PC). Cells were isolated by collagenase and pronase digestion followed by centrifugal elutriation. The freshly isolated cells were incubated in the presence of 5 mM glucose, 0.5 mM glutamine, 1 mM lactate, and 0.4 mM palmitate, and the oxidation rate of individual substrates was determined by the measurement of 14CO2 production. Glucose utilization was assessed by detritiation of [2-3H]glucose. Glucose flux through HMS was 2.6, 1.6, and 0.72 nmol.h-1.mg protein-1 in KC, EC and PC, respectively. The oxidation rate of palmitate in PC (3.5 nmol.h-1.mg protein-1) was about twofold greater than in nonparenchymal cells. Glutamine oxidation was 6.1, 4.2, and 2.1 nmol.h-1.mg protein-1 in KC, EC, and PC, respectively. In contrast, oxidation of exogenous lactate by PC (32.1 nmol.h-1.mg protein-1) was about seven- to eightfold greater than by KC or EC. Presence of prevailing lactate concentrations did not inhibit glucose oxidation in these cells, while it attenuated glucose utilization by PC. Our data show that in the presence of a physiological substrate mixture, less than 20% of the ATP generated from exogenous substrates is derived from glycolysis in KC or EC. Oxidation of glutamine and palmitate are the main sources for energy in these cells. In PC, however, lactate and palmitate oxidation is responsible for approximately 90% for the ATP production derived form the oxidation of exogenous substrates.(ABSTRACT TRUNCATED AT 250 WORDS)
本研究评估并比较了库普弗细胞(KC)、内皮细胞(EC)和肝实质细胞(PC)中葡萄糖的利用速率、磷酸己糖途径(HMS)的活性以及谷氨酰胺、乳酸和棕榈酸的氧化情况。通过胶原酶和链霉蛋白酶消化,随后进行离心淘析来分离细胞。将新鲜分离的细胞在含有5 mM葡萄糖、0.5 mM谷氨酰胺、1 mM乳酸和0.4 mM棕榈酸的条件下孵育,并通过测量14CO2的产生来确定各个底物的氧化速率。通过[2-3H]葡萄糖的去氚化来评估葡萄糖的利用情况。在KC、EC和PC中,通过HMS的葡萄糖通量分别为2.6、1.6和0.72 nmol·h-1·mg蛋白-1。PC中棕榈酸的氧化速率(3.5 nmol·h-1·mg蛋白-1)比非实质细胞中的约高两倍。在KC、EC和PC中,谷氨酰胺氧化分别为6.1、4.2和2.1 nmol·h-1·mg蛋白-1。相比之下,PC对外源乳酸的氧化(32.1 nmol·h-1·mg蛋白-1)比KC或EC高约7至8倍。在这些细胞中,存在主要的乳酸浓度时,并不抑制葡萄糖氧化,而会减弱PC对葡萄糖的利用。我们的数据表明,在存在生理底物混合物的情况下,来自外源底物产生的ATP中,不到20%来自KC或EC中的糖酵解。谷氨酰胺和棕榈酸的氧化是这些细胞中的主要能量来源。然而,在PC中,乳酸和棕榈酸的氧化约占外源底物氧化产生ATP的90%。(摘要截取自250字)