TNF-α对SD大鼠胰岛细胞凋亡及信号传导影响的研究.pdf

The Study of Apoptosis and Insulin Signaling of Rat Pancreatic Islet Cells Caused by TNF-α Abstract Part One: The Effects of TNF-αon Rat Pancreatic Islet Cells Objective To investigate the role of TNF-α on apoptosis of rat pancreatic islet cells and expression of Bcl-2 and BaxmRNA. 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 o 7 days with RPMI 1640 medium and atmosphere of 95% air, 5%CO2 at 37 C. When the cells flatted to monolayer, they were divided into four groups and treated with different concentrations of TNF-α （0、10、30、50ng/mL ）for 24 hours respectively. The apoptosis of rat pancreatic islet cells was evaluated by TUNEL method, and the expression levels of Bcl-2 and BaxmRNA in these cells were measured by semiquantitive RT-PCR. Results Apoptotic numbers of pancreatic islet cells of 10ng/mLTNF-α group were not markedly higher than those of normal control group （P0.05 ）,but those of 30ng/mL and 50ng/mL group significantly increased than those of control group （P0.05 ）. The expression of Bcl-2mRNA slightly increased at concentration of 10ng/mL and then significantly decreased at 30-50ng/mL. Various concentration could promote expression of Bax mRNA. Conclusions (1) TNF-α could induce pancreatic islet cell apoptosis in vitro in dose-dependant manner. (2) 30ng/mL and 50ng/mLTNF-α could decrease expression of Bcl-2 mRNA of 4 pancreatic islet cells. 10ng/mL, 30ng/mL and 50ng/mLTNF-α could increase expression of BaxmRNA. That TNF-α could induce apoptosis of pancreatic