摘要
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 assess the effect of an overexpressed intracellular matrix metalloproteinase2 （MMP-2）on the INS-1 cells function.Methods Rat MMP-2 cDNA was cloned into eukaryotic plasmid pcDNA3.1 （＋） to construct MMP-2/pcDNA3.1（＋） plasmid.The cultured INS-1 cells were randomized to three groups：the control group,empty vector transfected group and MMP-2 transfected group,and then transfected by Lipofectmine 2000 to determine the expression of intracellular MMP-2 and the gelatinase activity of MMP-2.Apoptosis and insulin-secretion function in INS-1 cells after transfection was assessed as well.Results Higher expression of MMP-2 mRNA and protein,coming with up-regulation of its enzyme activities were found in MMP-2 transfected group,as compared to the control and vector transfected groups （P〈0.05）.The apoptosis rate was higher in MMP-2 transfected group[（56.07 ± 3,68） %] than in control group and empty vector transfected group[（33.70 ± 6.53） % and （38.02±5.60）% respectively,P〈0.05].However the insulin release index of MMP-2 transfected group was only（1,30 ± 0.27）,which was much lower as compared to control group （3.37 ± 0.76） and empty vector transfected group （2.90 ± 0.84） （P〈0.05）.Conclusion The overexpression of intracellular MMP-2 may be related with INS-1 cells apoptosis and with decreased pancreatic β cells function.
出处
《中国糖尿病杂志》
CAS
CSCD
北大核心
2014年第4期358-363,共6页
Chinese Journal of Diabetes
基金
国家自然科学基金(81170757)
上海交通大学医学院基金(09xj21011)
