Effects of ethanol on pancreatic beta-cell death: interaction with glucose and fatty acids.
Western lifestyle plays an important role in the prevalence of type 2 diabetes by causing insulin resistance and pancreatic beta-cell dysfunction, a prerequisite for the development of diabetes. High fat diet and alcohol are major components of the western diet. The aim of the present study was to investigate the effects of ethanol and fatty acids on beta-cell survival and metabolism. We treated the rat beta-cell line RINm5F with ethanol, a mixture of palmitic and oleic acids, or both. Reactive oxygen species (ROS) were determined by (5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate) (CM-H2DCFDA) fluorescence assay, and mitochondrial activity was assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) reduction assay and by determining ATP production. Cell viability was assessed with a cell counter and trypan blue exclusion, and the mode of cell death by Hoechst33342 and propidium iodide staining. With both ethanol and fatty acid treatments, MTT reduction and ATP production decreased, whereas ROS production increased. Ethanol treatment had no effect on cell number, whereas fatty acid treatment reduced the cell number. Cell incubation with ethanol, fatty acids, or both increased the number of Hoechst 33342-positive nuclei. However, the majority of nuclei from fatty acid-treated cells were stained with propidium iodide, indicating a loss of plasma membrane integrity. We conclude that both ethanol and fatty acids generate cellular oxidative stress, and affect mitochondrial function in RINm5F beta-cells. However, ethanol causes beta-cell death by apoptosis, whereas fatty acids cause cell death predominantly by necrosis. It is not known whether these results are applicable to human beta-cells.
Dembele
Korami
K
Department of Internal Medicine, University of Manitoba, Winnipeg, Manitoba, Canada.
Nguyen
K Hoa
KH
Hernandez
Tiffany A
TA
Nyomba
B L Grégoire
BL
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D002492:Central Nervous System Depressants / Q000494:pharmacology*; D003924:Diabetes Mellitus, Type 2 / Q000378:metabolism / Q000473:pathology; D004347:Drug Interactions; D000431:Ethanol / Q000494:pharmacology*; D005947:Glucose / Q000494:pharmacology*; D050417:Insulin-Secreting Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D019301:Oleic Acid / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects; D019308:Palmitic Acid / Q000494:pharmacology*; D051381:Rats
D002492:Central Nervous System Depressants; D019301:Oleic Acid; D019308:Palmitic Acid; D000431:Ethanol; D005947:Glucose