高糖对MMP-2与TIMP-2的表达及血管平滑肌细胞增殖的影响

目的探讨高糖作用下大鼠胸主动脉平滑肌细胞基质金属蛋白酶-2(MMP-2)及其组织抑制剂-2(TIMP-2)的表达情况及高糖对血管平滑肌细胞(VSMCs)增殖的影响。方法大鼠胸主动脉平滑肌细胞培养及传代后,分为正常浓度葡萄糖组(NG组,5.5mmol/L葡萄糖)、甘露醇高渗对照组(5.5mmol/L葡萄糖+19.5mmol/L甘露醇)、高浓度葡萄糖组(HG组,25mmol/L葡萄糖)和GM6001组(25mmol/L葡萄糖+5μmol/L GM6001),在4组不同的培养环境下分别培养72h后,用RT-PCR技术检测MMP-2与TIMP-2的表达情况,同时用MTT比色法检测4种不同培养环境下VSMCs的增殖情况。结果 HG组与NG组相比,MMP-2与TIMP-2mRNA的表达增强,差异具有统计学意义(P<0.01),甘露醇高渗对照组、GM6001组与NG组相比,MMP-2与TIMP-2mRNA的表达差异无统计学意义(P>0.05);HG组与NG组相比,MMP-2与TIMP-2 mRNA的相对表达量的比值显著增加,差异具有统计学意义(P<0.01),甘露醇高渗对照组、GM6001组与NG组相比差异无统计学意义(P>0.05);HG组各时间点细胞增殖与NG组相比差异具有统计学意义(P<0.05),而GM6001组、甘露醇高渗对照组各时间点的细胞增殖与NG组相比差异无统计学意义(P>0.05)。结论高浓度葡萄糖促进血管平滑肌细胞增殖,这可能是通过MMP-2与TIMP-2表达失衡介导的。

Effects of high glucose on MMP-2 and TIMP-2 expressions and proliferation of vascular smooth muscle cells

Objective To investigate the effect of high glucose on expressions of matrix metalloproteinase-2(MMP-2) and tissue inhibitor of metalloproteinase-2(TIMP-2) as well as on the proliferation of vascular smooth muscle cells(VSMCs).Methods After being cultured and passaged,the VSMCs were divided into four groups and cultured in four different culture conditions for 72 hours,respectively:normal glucose group(NG group,5.5 mmol/L Glu),mannitol group(5.5 mmol/L Glu + 19.5 mmol/L mannitol),high glucose group(HG group,25 mmol/L Glu) and GM6001 group(25 mmol/L Glu + 5 μmol/L GM6001).Then,the expressions of MMP-2 and TIMP-2 were detected by RT-PCR technique,and the proliferation of VSMCs was evaluated by MTT method in four culture conditions.Results The expressions of MMP-2 and TIMP-2 mRNA were significantly higher in HG group than in NG group(P<0.01).Compared with that in NG group,the ratio of relative expression of MMP-2 and TIMP-2 mRNA in HG group was also significantly increased(P<0.01).However,mannitol or GM6001 group did not differ significantly from NG group in the expressions or the ratio of relative expression of MMP-2 and TIMP-2 mRNA(P>0.05).The proliferation of VSMCs increased significantly at different time points in HG group as compared with NG group(P<0.05),but mannitol or GM6001 group did not significantly differ from NG group respectively(P>0.05).Conclusion High glucose promotes proliferation of VSMCs,which may be mediated through the imbalance of MMP-2 and TIMP-2 expressions.