Mechanisms mediating cephaloridine inhibition of renal gluconeogenesis

• Published in: Toxicology and applied pharmacology Vol. 87; no. 2; p. 297
• Main Authors: Goldstein, R S, Contardi, L R, Pasino, D A, Hook, J B
• Format: Journal Article
• Language: English
• Published: United States 01.02.1987
• Subjects: Animals; Cephaloridine - pharmacology; Fructose-Bisphosphatase - metabolism; Gluconeogenesis - drug effects; Glucose-6-Phosphatase - metabolism; Kidney - drug effects; Kidney - metabolism; Kidney Cortex - drug effects; Kidney Cortex - enzymology; Kinetics; Male; Pyruvates - pharmacology; Pyruvic Acid; Rats; Rats, Inbred F344; Substrate Specificity; Time Factors
• ISSN: 0041-008X
• DOI: 10.1016/0041-008x(87)90291-2

Incubation of renal cortical slices with cephaloridine (CPH) markedly inhibits pyruvate-supported gluconeogenesis, an effect which is independent of CPH-induced lipid peroxidation. CPH was found to inhibit pyruvate-supported gluconeogenesis in a time-and concentration-dependent manner. Pyruvate-supported gluconeogenesis was inhibited as early as 10 min following incubation of renal cortical slices with 5 mM CPH. Similarly, endogenous gluconeogenesis was impaired following CPH treatment. CPH depressed the renal cortical slice content of ATP by 50%, but only following 90 and 120 min of drug exposure, suggesting that mitochondrial dysfunction does not mediate the inhibition of gluconeogenesis by CPH. To identify the intracellular site(s) of CPH inhibition of gluconeogenesis, the effects of CPH on glucose production were evaluated using substrates catalyzed by rate-limiting reactions. CPH inhibited renal cortical slice gluconeogenesis when the following substrates were used: pyruvate (mitochondrial), oxaloacetate and fructose-1,6-diphosphate (FDP) (postmitochondrial), and glucose-6-phosphate (G6P, endoplasmic reticulum). Inhibition of G6P-supported gluconeogenesis occurred within 5 min of incubation with 5 mM CPH. Direct addition of CPH to microsomal suspensions inhibited G6Pase activity in a concentration-dependent fashion. By contrast, addition of CPH to cytosolic fractions did not affect FDPase activity. CPH increased the Km and decreased the Vmax of G6Pase, indicating mixed competitive and noncompetitive inhibition. These data indicate that the profound inhibition of renal cortical slice gluconeogenesis by CPH is mediated by inhibition of microsomal G6Pase activity.