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目的观察亚砷酸钠(NaAsO2)对胰岛β细胞NIT-1胰岛素合成与分泌的影响。方法体外培养NIT-1细胞,NaAsO2处理后,以MTT法检测细胞的增殖活性,用ELISA测定胰岛素分泌情况,RT-PCR法检测胰岛素mRNA的表达。结果 (1)当NaAsO2浓度大于4μmol/L时抑制细胞增殖,细胞增殖抑制率随NaAsO2浓度的增加及作用时间的延长而增加。(2)作用24h,8μmol/L组不同浓度葡萄糖刺激的胰岛素分泌和胰岛素mRNA表达均明显减少(P<0.05);作用48h,4μmol/L组和8μmol/L组不同浓度葡萄糖刺激的胰岛素分泌和胰岛素mRNA表达均明显减少(P<0.05),而且8μmol/L组对高糖(16.5mmol/L)刺激的胰岛素分泌与基础(5.5mmol/L)胰岛素分泌差异无统计学意义。结论砷(As)对NIT-1细胞胰岛素分泌和胰岛素基因表达的影响与作用时间和剂量有关,胰岛素合成与分泌障碍可能是(As)影响胰岛β细胞功能进而引发糖尿病的机制之一。
Objective To observe the effect of sodium arsenite (NaAsO2) on the insulin synthesis and secretion of pancreatic β-cell NIT-1. Methods NIT-1 cells were cultured in vitro. The proliferation of NIT-1 cells was detected by MTT assay after NaAsO2 treatment. The insulin secretion was measured by ELISA and the mRNA expression of insulin was detected by RT-PCR. Results (1) When the concentration of NaAsO2 was greater than 4μmol / L, the cell proliferation was inhibited. The inhibition rate of cell proliferation increased with the concentration of NaAsO2 and the extension of time. (2) 24h and 8μmol / L groups of different concentrations of glucose-stimulated insulin secretion and insulin mRNA expression were significantly reduced (P <0.05); 48h, 4μmol / L group and 8μmol / L group of different concentrations of glucose-stimulated insulin secretion and (P <0.05). There was no significant difference in the insulin secretion between the high glucose (16.5mmol / L) and basal (5.5mmol / L) insulin in the 8μmol / L group. Conclusion The effect of As on the insulin secretion and insulin gene expression in NIT-1 cells is related to the time and dose. Insulin synthesis and secretion disorders may be one of the mechanisms that affect the function of (β) islet β cells and then trigger diabetes mellitus.