title: Effects of insulin, biguanide antihyperglycaemic agents and beta-adrenergic agonists on pathways of myocardial proteolysis.
abstract: Pathways of bulk protein degradation controlled by insulin and isoprenaline (isoproterenol) were distinguished in Langendorff-perfused rat hearts. Proteins were biosynthetically labelled in vitro with [3H]leucine, followed by addition of 2 mM non-radioactive leucine to competitively prevent reincorporation. Rapidly degraded proteins were eliminated during a 3 h preliminary perfusion period without insulin. One third of bulk myocardial protein degradation was inhibited by isoprenaline as described previously. An insulin concentration of 5 nM maximally inhibited proteolysis, beginning within 2 min. Inhibition reached 32% within 1.25 h and 35% after 1.5 h. The minimum effective insulin concentration was approx. 10-50 pM, which caused 10-20% inhibition. Following 3 h of perfusion without insulin, the lysosomal inhibitor, chloroquine (30 microM), inhibited 38% of bulk degradation. The 35% proteolytic inhibition caused by insulin was followed by very little further inhibition on subsequent concurrent infusion of chloroquine, i.e. the inhibitory effects of insulin and chloroquine were not additive. In contrast, prior inhibition of lysosomal proteolysis by insulin or chloroquine did not prevent the subsequent additive inhibition caused by isoprenaline. Insulin and beta-agonists additively inhibited approx. two-thirds of bulk degradation. The biguanide antihyperglycaemic agent phenformin (2 microM) inhibited 35% of bulk degradation, beginning at 2 min and reaching a near maximum at approx. 1.25-1.5 h. Following inhibition of proteolysis with phenformin (20 microM), subsequent infusion of chloroquine (30 microM) produced only a slight additional inhibition. Following inhibition of 35% of degradation by 1.5 h of perfusion with insulin (5 nM), subsequent exposure to phenformin (2 microM) produced only a slight additional inhibition which did not exceed 38% of basal proteolysis. Thus insulin and phenformin both inhibit lysosomal proteolysis; however, the adrenergic-responsive pathway is distinct.
authors.affiliation: Department of Pharmacology and Toxicology, School of Medicine, Wright State University, Dayton, OH 45435.
authors.forename: D P
authors.initials: DP
authors.lastname: Thorne
authors.forename: T D
authors.initials: TD
authors.lastname: Lockwood
mesh_terms: D000318:Adrenergic beta-Agonists / Q000494:pharmacology*; D000818:Animals; D001645:Biguanides / Q000494:pharmacology*; D002738:Chloroquine / Q000494:pharmacology; D004359:Drug Therapy, Combination; D066298:In Vitro Techniques; D007328:Insulin / Q000494:pharmacology*; D007545:Isoproterenol / Q000494:pharmacology; D008247:Lysosomes / Q000201:enzymology / Q000378:metabolism*; D009200:Myocardial Contraction / Q000187:drug effects; D009206:Myocardium / Q000201:enzymology / Q000378:metabolism*; D010629:Phenformin / Q000494:pharmacology; D011480:Protease Inhibitors / Q000378:metabolism*; D011489:Protein Denaturation / Q000187:drug effects; D051381:Rats
chemical_list: D000318:Adrenergic beta-Agonists; D001645:Biguanides; D007328:Insulin; D011480:Protease Inhibitors; D002738:Chloroquine; D010629:Phenformin; D007545:Isoproterenol