TNF-α对SD大鼠胰岛细胞凋亡及信号传导影响的研究-临床医学专业论文.docxVIP

PAGE PAGE 10 pancreatic islet cells. 10ng/mL, 30ng/mL and 50ng/mLTNF-α could increase expression of BaxmRNA. That TNF-α could induce apoptosis of pancreatic islet cells might relate decrease of Bcl-2 mRNA expression and increase of BaxmRNA expression. Key word TNF-α，pancreatic islet cells，apoptosis，Bax， Bcl-2 Part Two The Influence of TNF-α on Insulin Signaling in Isolated Rat Pancreatic Islet Cells Objective To explore the influence of TNF-??on the expression of Insulin receptor substrate-1（IRS-1）、Insulin receptor substrate-2（IRS-2）and Glucose transporter-2 （Glut-2）mRNA in primary cultured rat pancreatic islet cells. Methods Pancreatic islet cells were obtained by Collagenase P digestion and Ficoll-400 density gradient purification from Adult SD rat pancreas. The islet cells were then cultured for 7 days with RPMI 1640 medium and atmosphere of 95% air, 5%CO2 at 37oC. When the islet cells flatted to monolayer, they were divided into four groups and incubated with different concentrations of TNF-α（0、10、30、50ng/mL respectively). The levels of insulin in supernatant of cultured islet cells of various group under the basal and glucose-stimulated conditions were studied by RIA, and the expression of IRS-1、IRS-2 and Glut-2 mRNA was then measured by semiquantitive RT-PCR. Results Insulin release ,both the basal and glucose-stimulated,could be significantly inhibited by addition of TNF-α(10ng/mL、30ng/mL and 50ng/mL) . Expression levels of IRS-1 and IRS-2mRNA of 10ng/mLTNF-α group were not significantly different from those of control group (P0.05) . 30ng/mL and 50ng/mL could significantly decrease expression levels of IRS-1 and IRS-2 mRNA. 10ng/mL、30ng/mL and 50ng/mLTNF-α could inhibit expression of Glut-2mRNA of pancreatic islet cell in dose-dependant manner. Conclusions Various concentration（10ng/mL, 30ng/mL, 50ng/mL）could inhibit insulin release of pancreatic islet cells under the basal and higher glucose-stimulated condi