Characterization of peripheral benzodiazepine receptors in purified large mammal pancreatic islets

Istituto Policattedra di Discipline Biologiche, Università di Pisa, Via Bonanno 6, 56126, Pisa, Italy
Biochemical Pharmacology (Impact Factor: 4.65). 06/1996; DOI: 10.1016/0006-2952(96)00082-2

ABSTRACT In this work, we evaluated the biochemical properties of peripheral benzodiazepine receptors (PBRs) in the porcine endocrine pancreas and their role in insulin release. Binding of [3H]1-(2-chlorophenyl-N-methyl-1-methyl-propyl)-3-isoquinolinecarboxamide ([3H]PK-11195), a specific ligand of PBRs, to islet membranes was saturable and Scatchard's analysis of saturation curve demonstrated the presence of a single population of binding sites, with a dissociation constant (Kd) value of 4.75 ± 0.70 nM and a maximum amount of specifically bound ligand (Bmax) of 4505 ± 502 fmol/mg of proteins. The pharmacological profile of PBRs was determined as the ability of PK-11195 and several benzodiazepine compounds to displace [3H]PK-11195 from these binding sites. The rank order of potency yielded the following affinity results: PK-11195 > 7-chloro-1,3-dihydro-1-methyl-5-(p-chlorophenyl)-2H-1,4-benzodiazepine-2-on (Ro 5-4864) > diazepam ⩾ flunitrazepam ⪢ flumazenil. Secretion studies demonstrated that PK-11195 (1 and 10 μM) and Ro 5-4864 (10 and 50 μM) significantly potentiated insulin secretion from freshly isolated porcine islets at 3.3 mM glucose. This potentiating effect was not observed at 16.7 mM glucose concentration nor by the addition of clonazepam. These results show the presence of PBRs in purified porcine pancreatic islets and suggest an implication of PBRs in the mechanisms of insulin release.

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    • "to PBR such as regulation of steroid production (Papadopoulos, 1993), inflammatory response (Torres et al, 1999), insulin secretion (Marchetti et al, 1996b), mitochondrial respiration (Hirsch et al, 1989; Krueger, 1995), cell differentiation (Canat et al, 1993) and cell proliferation. Proliferation of various tumours including breast cancer (Beinlich et al, 1999; Carmel et al, 1999), melanoma (Landau et al, 1998), testis (Garnier et al, 1993) and astrocytoma (Neary et al, 1995) was shown to be inhibited by PBR ligands at micromolar concentrations. "
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