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目的探讨细胞内基质金属蛋白酶2(MMP-2)基因过表达对大鼠胰岛β细胞株INS-1细胞功能的影响。方法采用基因重组技术将大鼠MMP-2cDNA插入真核表达载体pcDNA 3.1(+)构建大鼠MMP-2真核表达质粒,培养INS-1细胞。随机分为正常对照组,空质粒转染组及MMP-2质粒转染组。脂质体Lipofectmine 2000转染INS-1细胞,观察INS-1细胞内MMP-2基因和蛋白表达量变化,MMP-2酶活性变化,以及INS-1细胞凋亡情况与胰岛素分泌功能的变化。结果与正常对照组、空质粒转染组比较,MMP-2质粒转染组MMP-2mRNA表达增加(P均<0.05),蛋白表达水平和酶活性上调(P均<0.05);MMP-2质粒转染组细胞凋亡率(56.07±3.68)%高于正常对照组(33.70±6.53)%及空质粒转染组(38.02±5.60)%(P<0.05),而IRI(1.30±0.27)低于正常对照组(3.37±0.76)与空质粒转染组(2.90±0.84)(P<0.05)。结论细胞内MMP-2基因的过表达可引起胰岛β细胞凋亡增加,胰岛素分泌功能下降。
Objective To investigate the effect of overexpression of matrix metalloproteinase-2 (MMP-2) gene on the function of rat pancreatic β-cell INS-1 cells. Methods Rat MMP-2 cDNA was inserted into eukaryotic expression vector pcDNA 3.1 (+) by gene recombination technique to construct rat eukaryotic expression plasmid of MMP-2 and cultured INS-1 cells. Randomly divided into normal control group, empty plasmid transfected group and MMP-2 plasmid transfected group. Lipofectmine 2000 was transfected into INS-1 cells to observe the changes of MMP-2 gene and protein expression, the changes of MMP-2 enzyme activity, the changes of INS-1 cells apoptosis and insulin secretion in INS-1 cells. Results Compared with normal control group and empty plasmid transfected group, the expression of MMP-2 mRNA increased (P <0.05) and the protein level and enzyme activity increased (P <0.05) (56.07 ± 3.68)% in the transfected group was significantly higher than that in the control group (33.70 ± 6.53)% and empty plasmid transfected group (38.02 ± 5.60)% (P <0.05), while the IRI (1.30 ± 0.27) In normal control group (3.37 ± 0.76) and empty plasmid transfected group (2.90 ± 0.84) (P <0.05). Conclusion Overexpression of intracellular MMP-2 gene leads to the increase of pancreatic β-cell apoptosis and the decrease of insulin secretion.