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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 mellitus is a chronic metabolic disease that occurs when the islet β-cell function is impaired or diminished in number. This study mainly focused on the involvement of early growth responsive gene-1 (Egr-1) in β-cell apoptosis induced by high glucose. Using immortalized NIT-1 islet β cell line as experimental model, the cells were divided into normal control group and high glucose group. Cell viability and apoptosis were detected by MTT, DAPI staining, DNA Ladder and flow cytometry The transcription and protein expression of Egr-1 were detected by Real-time PCR and Western blot. The activity of Egr-1 was detected by competitive inhibition of Egr-1 activity and apoptosis. The results showed that compared with the control group, cell viability decreased and apoptosis increased markedly (48 h, P <0.05; 72 h, P <0.01). The transcription and protein expression of Egr-1 were significantly up-regulated P <0.01). Inhibition of its expression activity could significantly improve cell viability and decrease apoptosis (48 h, P <0.05). Suggesting that Egr-1 is indeed involved in high glucose-induced beta-cell apoptosis.