title: Pyroglutamyl-phenylalanyl-proline amide attenuates thyrotropin-releasing hormone-stimulated insulin secretion in perifused rat islets and insulin-secreting clonal beta-cell lines.
abstract: TRH immunoreactivity has been detected in the pancreas of man and rat and localized to the islets of Langerhans. We studied the effect of synthetic TRH and the related tripeptide pyroglutamyl-phenylalanyl-proline amide (EFP) on isolated perifused rat islets and the glucose-responsive clonal cell lines HIT-T15 and RIN5AH. TRH at 10 nM potentiated [0.5 +/- 0.1 (control) vs. 0.8 +/- 0.1 (TRH) pmol/10(6) cells per 120 min; mean +/- SEM; n = 6; P < 0.001; n = 15], whereas EFP from 1 nM upwards suppressed glucose-stimulated insulin secretion [0.8 +/- 0.1 (control) vs. 0.5 +/- 0.1 (EFP) pmol/10(6) cells per 120 min; P < 0.001; n = 12) in the cell lines. Further, EFP reversed TRH-stimulated insulin release. Similar responses were observed in perifused isolated rat islets at the tested dose of 1 microM. Gel permeation chromatography of rat adult and neonatal whole pancreas, isolated islets, and HIT cell extracts demonstrated the elution of total TRH-like immunoreactivity (t-TRH-LI) in the same position as synthetic TRH. Cation exchange analysis of the t-TRH-LI from rat adult pancreas and HIT cell extracts showed that neutral TRH-like peptides corresponding to synthetic EFP were also present. Reverse-phase fast protein liquid chromatographic analysis of t-TRH-LI in the unbound fraction of these extracts subjected to anion exchange columns, also demonstrated peaks corresponding to synthetic EFP. We conclude that TRH potentiates, whereas EFP inhibits, glucose-stimulated insulin release in isolated perifused islets and the cell lines. In addition, EFP reversed the stimulatory effect of TRH. The presence of EFP-LI in rat adult and neonatal pancreas and HIT cell extracts suggests it may contribute in the modulation of pancreatic endocrine function.
authors.affiliation: Francis Fraser Laboratories, Department of Metabolic Medicine, Royal Postgraduate Medical School, Hammersmith Hospital, London, United Kingdom.
authors.forename: R N
authors.initials: RN
authors.lastname: Kulkarni
authors.forename: Z L
authors.initials: ZL
authors.lastname: Wang
authors.forename: K O
authors.initials: KO
authors.lastname: Akinsanya
authors.forename: W M
authors.initials: WM
authors.lastname: Bennet
authors.forename: R M
authors.initials: RM
authors.lastname: Wang
authors.forename: D M
authors.initials: DM
authors.lastname: Smith
authors.forename: M A
authors.initials: MA
authors.lastname: Ghatei
authors.forename: P G
authors.initials: PG
authors.lastname: Byfield
authors.forename: S R
authors.initials: SR
authors.lastname: Bloom
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D006224:Cricetinae; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D007295:Inositol Phosphates / Q000378:metabolism; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007340:Insulinoma; D007515:Islets of Langerhans / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000494:pharmacology*; D010190:Pancreatic Neoplasms; D010477:Perfusion; D011761:Pyrrolidonecarboxylic Acid / Q000031:analogs & derivatives; D051381:Rats; D017208:Rats, Wistar; D013973:Thyrotropin-Releasing Hormone / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
chemical_list: D007295:Inositol Phosphates; D007328:Insulin; D009842:Oligopeptides; C074033:pyroglutamyl-phenylalanyl-prolinamide; D013973:Thyrotropin-Releasing Hormone; D005947:Glucose; D011761:Pyrrolidonecarboxylic Acid