高浓度葡萄糖对胰岛细胞凋亡和GLUT2、IRS-1、IRS-2+mRNA表达影响地.pdf

The Research of Various High-dose Glucose cause Cell Apoptosis and change GLUT2/IRS-1/IRS-2 mRNA Expression in Isolated Rat Pancreatic Islet Cells Abstract Part one The mechanism of various high-dose glucose cause cell apoptosis in isolated rat pancreatic islet cells Objective To research the effects of various high-dose glucose cause cell apoptosis in isolated rat pancreatic islet cells. Methods Islet cells were obtained by Collagenase P digestion and Ficoll-400 density gradient purification from Adult SD rat pancreas, which was cultured in RPMI-1640 culture media for 2 days.Then they were treated with various high-dose glucose for 5 days. The insulin concentrations in the supernatant were measured by Radioimmuno- assay (RIA) . The morphology of cells were observed when they were stained by AO/EB. Cell apoptosis rates were calculated by Tunel Kit. Total RNA of islets were abstracted by TRIzol reagent. The levels of bcl-2 and bax mRNA were measured by reverse transcription polymerase chain reaction (RT-PCR). Results Insulin concentrations increased in 11.1mmol/L glucose group and 22.2 mmol/L glucose group (P0.05), but there were no difference between them(P 0.05). Insulin concentration decreased in 26.7mmol/L glucose group. High dose glucose caused islets apoptosis.There were islet necrosis in 26.7mmol/L glucose group. Decreased bcl-2 level and increased bax level were detected in three high-dose glucose groups(P0.05),but there were no difference among them(P 0.05). Conclusions （1）、Islets apoptosis increased in 11.1mmol/L glucose group,22.2 mmol/L glucose group and 26.7mmol/L glucose group. （2 ）、Islets necrosis was detected in 26.7mmol/L glucose group. （3）、The mechanism of apoptosis in high-dose glucose may be related to the increasin