Glycated fetal calf serum affects the viability of an insulin-secreting cell line in vitro
| Autor: | Giorgio Luciano Viviani, Patrizio Odetti, Alessandra Puddu, Anna Garuti, Gianni Sacchi, Arianna Durante, Fiammetta Monacelli, Daniela Storace |
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| Rok vydání: | 2008 |
| Předmět: |
Glycation End Products
Advanced medicine.medical_specialty Cell Survival Endocrinology Diabetes and Metabolism medicine.medical_treatment Apoptosis Biology Arginine medicine.disease_cause Thiobarbituric Acid Reactive Substances Cell Line chemistry.chemical_compound Endocrinology Glycation Insulin-Secreting Cells Internal medicine Insulin Secretion medicine Animals Humans Insulin Pentosidine Cell Proliferation Cell growth Lysine Age Factors Oxidative Stress chemistry Cell culture Cattle Intracellular Oxidative stress |
| Zdroj: | Metabolism. 57:163-169 |
| ISSN: | 0026-0495 |
| DOI: | 10.1016/j.metabol.2007.08.020 |
| Popis: | The purpose of the present study was to evaluate the direct effects of advanced glycation end products (AGEs) on beta-cells by their exposure to a glycated serum to estimate the cellular viability and the related insulin secretion. Glycation of fetal calf serum was obtained by incubation with 50 mol/L ribose at 37 degrees C for 7 days; at the end of this incubation period, the pentosidine content ranged between 15 and 16 x 10(5) pmol/L. HIT-T15 cells, a pancreatic islet cell line, were grown and cultured for 5 days in Roswell Park Memorial Institute (RPMI) medium containing either not glycated (NGS) or glycated (GS) fetal calf serum. Cellular oxidative stress (ie, thiobarbituric acid-reactive substances) was assessed by high-performance liquid chromatography. Cellular viability was evaluated by detection of proliferation, cell necrosis, and cell apoptosis rate. The insulin secretion and the related intracellular content were evaluated by enzyme-linked immunosorbent assay. The present study reported, after 5 days of exposure to the glycation environment, a moderately reduced cellular proliferation (-20.44% +/- 2.92%) with a corresponding increase of cell necrosis (+67.7% +/- 1.56%) and cell apoptosis (+39.83% +/- 2.92%) rate in comparison with the untreated cells. Oxidative intracellular stress was higher in GS conditions compared with the NGS ones (+293.3% +/- 87.53%). Insulin release from GS-treated HIT-T15 cells was lower than that of NGS-treated cells both when cells were stimulated with low glucose concentration (2.8 mmol/L, -30.3% +/- 4.91%) or when they were challenged with high glucose concentration (16.7 mmol/L, -29.2% +/- 5.82%). Incubation of HIT-T15 cells with glycated serum also caused a significant decrease of insulin intracellular content (-44.47% +/- 9.98%). Thus, AGEs were shown to exert toxic effects on insulin-secreting cells. Chronically high intracellular oxidative stress, due to accumulation of AGEs, affects the insulin secretion machinery. The present data suggest a pivotal role of the non-enzymatic glycation process in the onset and progression of diabetes during aging and a direct adverse effect of a glycated environment on the pancreatic islet cells. |
| Databáze: | OpenAIRE |
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