Attenuation of Expression of γ-Glutamylcysteine Synthetase by Ribozyme Transfection Enhance Insulin Secretion by Pancreatic β Cell Line, MIN6
Low levels of intracellular antioxidant enzyme activities as well as glutathione (GSH) concentrations have been described in pancreatic β cells. We examined the effects of intracellular GSH depletion on insulin secretion and the role of intracellular GSH in signal transduction in β cell line, MIN6 c...
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Veröffentlicht in: | Biochemical and biophysical research communications 2000-11, Vol.278 (1), p.236-240 |
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Zusammenfassung: | Low levels of intracellular antioxidant enzyme activities as well as glutathione (GSH) concentrations have been described in pancreatic β cells. We examined the effects of intracellular GSH depletion on insulin secretion and the role of intracellular GSH in signal transduction in β cell line, MIN6 cells. Anti-γ-glutamylcysteine synthetase (γ-GCS) heavy subunit ribozyme was stably transfected to MIN6 cells to reduce intracellular GSH concentration. In the presence of 10 mM glucose, ribozyme-transfected cells (RTC) increased insulin secretion from 0.58 μg/106 cells/h in control cells (CC) to 1.48 μg/106 cells/h. This was associated with increased intracellular Ca2+ concentration in RTC, detected by fluo-3 staining. Our results demonstrated that intracellular GSH concentration might influence insulin secretion by MIN6 cells, and suggest that enhanced insulin secretion by β cells conditioned by chronic depletion of GSH is mediated by increased intracellular Ca2+ concentration. |
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ISSN: | 0006-291X 1090-2104 |
DOI: | 10.1006/bbrc.2000.3776 |