摘要
糖尿病是一种慢性代谢性疾病,胰岛β细胞功能损伤或数量减少都会导致其发生。该研究主要探讨早期反应基因-1(early growth responsive gene-1,Egr-1)参与高糖诱导β细胞凋亡的过程。利用永生化NIT-1胰岛β细胞株为实验模型,分成正常对照组和高糖组;通过MTT、DAPI染色和DNA Ladder以及流式细胞术等手段检测持续性高糖刺激对细胞活力和凋亡的影响;采用Real-time PCR和Western blot法检测Egr-1转录和蛋白表达水平;竞争性抑制Egr-1活性后检测细胞活力和凋亡的变化。研究发现,与对照组相比持续性高糖刺激后细胞活力下降,凋亡明显增加(48 h,P<0.05;72 h,P<0.01);Egr-1转录和蛋白表达水平均显著上调(P<0.01),抑制其表达活性可以明显改善细胞活力和降低凋亡水平(48 h,P<0.05)。提示Egr-1的确参与到高糖诱导β细胞凋亡的过程中。
Diabetes is a chronic metabolic syndrome caused by the impaired function or reduced amount of pancreas beta cells. Herein, we studied the detailed mechanism by which the early growth responsive gene-1 （Egr-1） was involved in the high glucose induced apoptosis of beta ceils. Using NIT-1 ceils as a cell model, we measured the effect of consistent high glucose on the cell viability and apoptosis through MTT, DAPI, DNA Ladder and flow cytometry by comparing the control group with the high glucose group. The Egr-1 expression level was determined by Real-time PCR and Western blot. We also detected the cell viability and apoptosis after inhibiting the Egr-1 function. We found that the high glucose treatment decreased the cell viability and increased the cell apoptosis significantly （48 h, P〈0.05; 72 h, P〈0.01）. Egr-1 was highly expressed （P〈0.01）. Inhibiting the Egr-1 function could improve the cell viability and reduce apoptosis （48 h, P〈0.05）. These results suggested that Egr-1 was involved in the high glucose induced beta cell apoptosis.
出处
《中国细胞生物学学报》
CAS
CSCD
北大核心
2013年第7期916-921,共6页
Chinese Journal of Cell Biology
