Melatonin suppresses cyclosporine A-induced autophagy in rat pituitary GH3 cells.
Cyclosporine A (CsA) is a powerful immunosuppressive drug with side effects including the induction of chronic nephrotoxicity including endoplasmic reticulum (ER) stress in tubular cells. Recently, it was reported that autophagy is induced by ER stress and serves to alleviate the associated deleterious effects. In the current study, CsA treatment (0-100 microm) decreased cell survival of rat pituitary GH3 cells in a dose-dependent manner. At concentrations ranging from 1.0 to 10 microm, CsA induced a dose-dependent increase in the expression of microtubule-associated protein 1 light chain 3 (LC3)-I and LC3-II. Cells treated with 2.5 microm CsA exhibited cytoplasmic vacuolation, indicating that CsA induces autophagy in rat pituitary GH3 cells. In the presence of 1.0-10 microm CsA, the expression of catalase decreased while that of the ER stress markers, ER luminal binding protein (BiP) and inositol-requiring enzyme 1 alpha (IRE1alpha), increased as compared those levels in untreated cells. These results suggested that CsA-induced autophagy is dependent on ER stress. To determine whether melatonin would protect cells against CsA-induced autophagy, we treated rat pituitary GH3 cells with melatonin in the presence of CsA. Melatonin treatment (100 and 200 microm) suppressed autophagy induced by 2.5 and 5 microm CsA. Furthermore, co-treatment with 100 microm melatonin inhibited LC3-II expression, and increased catalase and phosphorylated p-ERK levels in the presence of 2.5 and 5 microm CsA. BiP and IRE1alpha expression in melatonin-co-treated cells was superior to that in cells treated with 2.5 and 5 microm CsA alone. Thus, melatonin suppresses CsA-mediated autophagy in rat pituitary GH3 cells.
Laboratory of Veterinary Biochemistry and Molecular Biology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk, Republic of Korea.|Department of Biomedical Engineering, College of Health Science, Yonsei University, Wonju, Kangwon-do, Republic of Korea.
Yeong-Min
YM
Yoo
Laboratory of Veterinary Biochemistry and Molecular Biology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk, Republic of Korea.
Eui-Bae
EB
Jeung
D000704:Analysis of Variance; D000818:Animals; D001343:Autophagy / Q000187:drug effects*; D015153:Blotting, Western; D002374:Catalase / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D016572:Cyclosporine / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D004347:Drug Interactions; D004721:Endoplasmic Reticulum / Q000378:metabolism; D000091342:Endoplasmic Reticulum Chaperone BiP; D004722:Endoribonucleases / Q000378:metabolism; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000378:metabolism; D008550:Melatonin / Q000494:pharmacology*; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D010902:Pituitary Gland / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D051381:Rats; D013312:Stress, Physiological; D051028:bcl-2-Associated X Protein / Q000378:metabolism
D002374:Catalase; D016572:Cyclosporine; D000091342:Endoplasmic Reticulum Chaperone BiP; D004722:Endoribonucleases; D048049:Extracellular Signal-Regulated MAP Kinases; D006360:Heat-Shock Proteins; D008550:Melatonin; D008869:Microtubule-Associated Proteins; D017346:Protein Serine-Threonine Kinases; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; C494828:Bax protein, rat; C526109:Anxa3 protein, rat