Long-term effects of glibenclamide on the insulin production, oxidative metabolism and quantitative ultrastructure of mouse pancreatic islets maintained in tissue culture at different glucose concentrations

Abstract

In order to evaluate long-term effects of sulphonylureas on pancreatic islet structure and function, isolated mouse islets were maintained in tissue culture for one week at various glucose concentrations, and in the absence or presence of glibenclamide. When the islets were cultured at 3.3 or 5.5 mmol/1, but not at 16.7 mmol/1 glucose, it was found that the drug stimulated insulin secretion into the culture medium during the initial 3 days of culture. During the remainder of the culture period no such enhancement of secretion was demonstrated. Insulin release due to glibenclamide apparently resulted in rapid depletion of intracellular insulin stores. The finding of an enlarged B-cell Golgi apparatus in the drug-treated islets was probably associated with granule discharge. The failure of glibenclamide to promote insulin secretion during the whole culture period could reflect the adverse effects of the drug on islet insulin biosynthesis as indicated by short-term experiments performed after culture. Similar experiments showed that the impaired insulin biosynthesis could not be restored by withdrawal of the drug from the culture medium for 3 days. Furthermore, the capacity for insulin release in response to an acute glucose challenge at the end of the culture period, was abolished by culture in the presence of glibenclamide. The drug effects on insulin biosynthesis and intracellular insulin stores, which were most pronounced at 5.5 mmol/1 glucose, possibly resulted from changes in B-cell metabolism as suggested by the diminished islet glucose-oxidation rate. The spatial characteristics of islet mitochondria indicated that these changes might involve an adaptation to substrates other than glucose. In conclusion, our findings suggest that sulphonylureas have an insulinotropic effect, which is however transient. Indeed, it rather seems as if long-term exposure of islet B-cells to sulphonylureasin vitro were accompanied by functional deficiency.