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体外低糖培养胰岛β细胞,以葡萄糖/葡萄糖氧化酶(G/GO)为过氧化氢(H2O2)产生体系摸拟体内氧化应激水平,以流式细胞术检测细胞内H2O2,以RT-PCR。方法检测胰岛β细胞内胰腺十二指肠同源异型盒(PDX-1)的mRNA表达,检测胰岛素和C肽浓度。结果随着细胞内产生的H2O2逐渐升高,PDX-1的mRNA的表达逐步减少,胰岛素及C肽量也逐渐减少,而加入过氧化氢酶(CAT)清除H2O2后可部分逆转这一趋势。结论氧化应激可减少胰岛β细胞内的PDX-1 mRNA表达,从而减少胰岛β细胞胰岛素和C肽的分泌。
The islet β cells were cultured in vitro with hypoglycemic glucose, and the in vivo oxidative stress was detected by using glucose / glucose oxidase (G / GO) as hydrogen peroxide (H2O2) generating system. The intracellular H2O2 was detected by flow cytometry. Methods The mRNA expression of PDX-1 in pancreatic β-cell was detected and the concentrations of insulin and C-peptide were measured. Results With the gradual increase of intracellular H2O2, the mRNA expression of PDX-1 gradually decreased and the amount of insulin and C-peptide decreased gradually. This phenomenon was partially reversed by the addition of catalase (CAT) to eliminate H2O2. Conclusion Oxidative stress can reduce the expression of PDX-1 mRNA in islet β cells, thereby reducing the secretion of insulin and C peptide in pancreatic β cells.