MIN6细胞中亲磷酸酯酶A1基因表达与胰岛素分泌关系的初步研究

The primary research of the relation between expression of phosphatidic acid-preferring phospholipase A1 gene and insulin secretion in MIN6 cells

目的研究亲磷酸酯酶A1(PA-PLA1)基因在小鼠分泌胰岛素细胞株M IN 6中的表达及其与胰岛素分泌的关系。方法对M IN 6细胞进行不同浓度和不同时间的葡萄糖刺激胰岛素分泌的实验,酶联免疫吸附法测定细胞上清液中胰岛素分泌量,实时荧光定量PC R检测相应条件下PA-PLA1 m R N A的表达水平,线性相关分析PA-PLA1 m R N A表达水平与胰岛素分泌的相关性。结果在一定葡萄糖浓度范围内,PA-PLA1m R N A水平随着葡萄糖浓度的升高和刺激时间的延长而升高,并与细胞胰岛素分泌量呈明显的正相关。不同葡萄糖浓度和不同刺激时间的PA-PLA1 m R N A水平与胰岛素水平的相关系数r分别为0.89(P<0.01)和0.98(P<0.01)。结论小鼠分泌胰岛素细胞株M IN 6中PA-PLA1基因的表达与胰岛素分泌之间具有高度的正相关性。

【Objective】 To investigate the relation between the expression of phosphatidic acid-preferring phospholipase A1（PA-PLA1） gene and the secretion of insulin in mouse insulin-secreting cell line MIN6.【Methods】 The glucose-stimulated insulin secretion（GSIS） experiment was carried out by stimulating MIN6 cells with glucose at various concentrations for 2 h or at the injection of 25 mmol/L for different durations.The PA-PLA1 mRNA level was determined by real-time quantitative PCR,and the insulin secretion was measured with enzyme linked immunosorbent assay.The relation between the levels of PA-PLA1 mRNA and insulin was analyzed by linear correlation.【Results】 In the GSIS at various concentrations,the PA-PLA1 mRNA level increased as the amout of glucose injection rising within the range of 5～25 mmol/L,however the PA-PLA1 mRNA level stimulated by 50 mmol/L glucose was lower than that by 25 mmol/L.Moreover,the change of insulin secretion was parallel to that of PA-PLA1 mRNA expression.The relation between the levels of PA-PLA1 mRNA and insulin was highly significant（r =0.89,P〈 0.01）.In the GSIS for different durations,the PA-PLA1 mRNA level increased with time and the insulin level showed a similar trend.The relation between them was also highly significant（r =0.98,P〈 0.01）.【Conclusion】 There is a highly positive correlation between the PA-PLA1 mRNA expression and insulin secretion in mouse insulin-secreting MIN6 cells.