| Objective: To investigate effects of high level of free fatty acid and glucose on islet cells apoptosis.Methods : SD rat pancreatic islets were isolated and divided into five groups, control group, glucose group 1, glucose group2,free fatty acid group land free fatty acid group 2,according to different cultured condition. After culturing 48 hours, then islets were incubated in low glucose(5.6mmol/l) and high glucose(25.6mmol/l) respectively for 1 hour , the insulin concentrations in the culture medium were detected by radio immunological methods; Apoptotic cells were identified by TUNEL method ;The expressions of caspase-3 and cytochrome C were evaluated by immunohistochemical techniques.Results : (1) insulin secretion decreased when islet cells were cultured with high glucose and free fatty acid for 48h: the insulin levels of high glucose group were significantly lower than those of control group (p <0.01) ,the insulin levels of the different free fatty acid groups were markedly lower than those of the control group ( p <0.01) , the insulin levels of the free fatty acid group 2 were significantly lower than those of the free acid group 1.(2) The apoptotic rates were increased when islet cells were cultured by high glucose and free fatty acid for 48 hours : the apoptotic rates of different high glucose groups and free fatty acid groups were markedly higher than those of the control group (p <0.01) . the apoptotic rates of the free fatty acid group 2 were markedly higher than those of the free fatty acid group 1 (p <0.01) .(3)The expressions of caspase-3 and cytochrome C increased when islet cells were cultured with high glucose and free fatty acid for 48 hours:The expressions of caspase-3 and cytochrome C in the different high glucose groups and free fatty acid groups were markedly higher than those in the control group (p <0.01) . the expressions of caspase-3 and cytochrome C in the free fatty acid group 2 were markedly higher than those in free fatty acid... |
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