title: Differential influence of tacrolimus and sirolimus on mitochondrial-dependent signaling for apoptosis in pancreatic cells.
abstract: To examine and compare the mitochondria-related cellular mechanisms by which tacrolimus (TAC) or sirolimus (SIR) immunosuppressive drugs alter the pancreatic exocrine and endocrine β-cell fate. Human exocrine PANC-1 and rat endocrine insulin-secreting RIN-m5F cells and isolated rat islets were submitted to 1-100 nM TAC or SIR. In cultures, insulin secretion was measured as endocrine cell function marker. Apoptosis was quantified by annexin 5 and propidium iodide staining. Cleaved caspase-3, Bax apoptosis indicators, and p53, p21 cell cycle regulators were detected by Western blot. Cell cycle and mitochondrial membrane potential (ΔΨm) were analyzed by flow cytometry and SA-beta-galactosidase (SA-β-gal) activity by fluorescence microscopy. Only TAC reduced insulin secretion by RIN-m5F after 24 h. TAC and SIR promoted moderate apoptosis in both PANC-1 and RIN-m5F after 24 h. Apoptosis was associated with up-regulated Bax (threefold) and cleaved caspase-3 (fivefold) but only in PANC-1, while p53 and p21 were up-regulated (twofold) in both cell lines. ΔΨm was impaired only in PANC-1 by TAC and SIR. Only SIR prompted cell cycle arrest in both cell lines. The induction of a premature senescence-like phenotype was confirmed in isolated islets by SA-β-gal activity. TAC and SIR are early inducers of pancreatic cell dysfunction and apoptosis but differentially alter endocrine and exocrine cells via mitochondrial-driven pathways. In rat islets, TAC and SIR prompt a senescence-like phenotype.
authors.affiliation: EA7293, Vascular and Tissular Stress in Transplantation, Federation of Translational Medicine of Strasbourg, Faculty of Medicine, University of Strasbourg, 74 route du Rhin, 67401, Illkirch, Strasbourg, France. andrei-alexandru.constantinescu@inserm.fr.|Department of Parasitology and Parasitic Diseases and Animal Biology, Faculty of Veterinary Medicine, University of Agronomical Sciences and Veterinary Medicine, 105 spl. Independentei, Sector 5, 050097, Bucharest, Romania. andrei-alexandru.constantinescu@inserm.fr.|INSERM, UMR866, «Equipe labellisée Ligue contre le Cancer » and Laboratoire d'Excellence LipSTIC, 21079, Dijon, France. andrei-alexandru.constantinescu@inserm.fr.
authors.forename: Andrei Alexandru
authors.initials: AA
authors.lastname: Constantinescu
authors.affiliation: EA7293, Vascular and Tissular Stress in Transplantation, Federation of Translational Medicine of Strasbourg, Faculty of Medicine, University of Strasbourg, 74 route du Rhin, 67401, Illkirch, Strasbourg, France.|Ecole Doctorale de Sciences et Technologies, Université Libanaise, Campus Universitaire de Rafic Hariri, Hadath, Lebanon.
authors.forename: Malak
authors.initials: M
authors.lastname: Abbas
authors.affiliation: EA7293, Vascular and Tissular Stress in Transplantation, Federation of Translational Medicine of Strasbourg, Faculty of Medicine, University of Strasbourg, 74 route du Rhin, 67401, Illkirch, Strasbourg, France.
authors.forename: Mohamad
authors.initials: M
authors.lastname: Kassem
authors.affiliation: EA7293, Vascular and Tissular Stress in Transplantation, Federation of Translational Medicine of Strasbourg, Faculty of Medicine, University of Strasbourg, 74 route du Rhin, 67401, Illkirch, Strasbourg, France.
authors.forename: Céline
authors.initials: C
authors.lastname: Gleizes
authors.affiliation: EA7293, Vascular and Tissular Stress in Transplantation, Federation of Translational Medicine of Strasbourg, Faculty of Medicine, University of Strasbourg, 74 route du Rhin, 67401, Illkirch, Strasbourg, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Kreutter
authors.affiliation: UMR7213 CNRS, Laboratory of Biophotonics and Pharmacology, 74 route du Rhin, 67401, Illkirch, France.
authors.forename: Valerie
authors.initials: V
authors.lastname: Schini-Kerth
authors.affiliation: Department of Parasitology and Parasitic Diseases and Animal Biology, Faculty of Veterinary Medicine, University of Agronomical Sciences and Veterinary Medicine, 105 spl. Independentei, Sector 5, 050097, Bucharest, Romania.
authors.forename: Ioan Liviu
authors.initials: IL
authors.lastname: Mitrea
authors.affiliation: UMR7213 CNRS, Laboratory of Biophotonics and Pharmacology, 74 route du Rhin, 67401, Illkirch, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Toti
authors.affiliation: EA7293, Vascular and Tissular Stress in Transplantation, Federation of Translational Medicine of Strasbourg, Faculty of Medicine, University of Strasbourg, 74 route du Rhin, 67401, Illkirch, Strasbourg, France. kesslerl@unistra.fr.|Department of Diabetology, University Hospital, 1 Place de l'Hôpital, Clinique Médicale B, CHU de Strasbourg, BP421, 67091, Strasbourg Cedex, France. kesslerl@unistra.fr.
authors.forename: Laurence
authors.initials: L
authors.lastname: Kessler
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002460:Cell Line; D006801:Humans; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000378:metabolism*; D046790:Pancreas, Exocrine / Q000378:metabolism*; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects*; D020123:Sirolimus / Q000494:pharmacology*; D016559:Tacrolimus / Q000494:pharmacology*
chemical_list: D020123:Sirolimus; D016559:Tacrolimus
keywords: Cellular premature senescence; Endocrine β-cells; Exocrine cells; Mitochondria-related apoptosis; Sirolimus; Tacrolimus