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目的 观察酒精对NIT - 1细胞葡萄糖刺激后胰岛素分泌功能及葡萄糖激酶 (Glucokinase)基因表达的影响。方法 体外培养NIT - 1细胞 ,使用不同浓度的酒精 (0 ,5 0 ,1 0 0 ,2 0 0 ,4 0 0mmol/L)作用不同时间后 (6 ,1 2 ,2 4h) ,用放射免疫法测定NIT - 1细胞分泌胰岛素的量 ,用噻唑蓝 (MTT)法检测NIT - 1细胞代谢活性 ,用RT PCR方法检测葡萄糖激酶基因mRNA的表达。结果 酒精对NIT - 1细胞代谢活性的抑制与酒精剂量和作用时间有关。酒精作用 6h ,NIT - 1细胞葡萄糖刺激的胰岛素分泌增加 ;作用 1 2 ,2 4h ,葡萄糖刺激的胰岛素分泌降低。酒精作用 6h ,各剂量组葡萄糖激酶mRNA表达升高 ;酒精作用 1 2 ,2 4h ,各剂量组葡萄糖激酶mRNA表达均降低。结论 酒精对NIT - 1细胞葡萄糖刺激后胰岛素分泌及葡萄糖激酶mRNA表达的影响与作用时间、剂量有关。而葡萄糖激酶mR NA表达水平的改变可能是酒精影响NIT - 1细胞胰岛素分泌的重要机制之一。
Objective To investigate the effects of alcohol on insulin secretion and glucokinase gene expression in NIT - 1 cells after glucose stimulation. Methods NIT - 1 cells were cultured in vitro. The cells were treated with different concentrations of alcohol (0, 500, 100, 200 mmol / L) for different time periods (6, 12, 24 hours) The amount of insulin secreted by NIT - 1 cells was measured. The metabolic activity of NIT - 1 cells was detected by MTT assay. The mRNA expression of glucokinase was detected by RT - PCR. Results The inhibitory effect of alcohol on the metabolic activity of NIT - 1 cells was related to the dose of alcohol and the duration of action. After 6 hours of alcohol treatment, the glucose stimulated insulin secretion increased in NIT - 1 cells. The glucose - stimulated insulin secretion decreased 12 days and 24 hours after treatment with alcohol. After 6 hours of alcohol administration, the expression of glucokinase mRNA in each dose group was increased. At the same time, the mRNA expression of glucokinase was decreased in each dose group at 12 and 24 hours. Conclusion The effect of alcohol on insulin secretion and glucokinase mRNA expression after glucose stimulation in NIT - 1 cells is related to time and dosage. The change of mR NA expression of glucose kinase may be one of the important mechanisms by which alcohol affects the insulin secretion of NIT - 1 cells.