同型半胱氨酸对A7r5细胞增殖及MMP-2表达的影响

    Influence of homocysteine on the proliferation and expression of matrix metalloproteinase 2 in A7r5 cells

    • 摘要: 目的:观察同型半胱氨酸(Hcy)对大鼠胸主动脉平滑肌细胞(A7r5细胞)增殖及基质金属蛋白酶2(MMP-2)表达的影响。方法:分别用0、0.25、0.5、1 mmol/L Hcy作用A7r5细胞48 h后,细胞计数及MTT法检测A7r5细胞的增殖情况,半定量逆转录聚合酶链反应(RT-PCR)检测MMP-2 mRNA的表达,免疫印迹法(Western blot)检测MMP-2蛋白的表达。结果:随着Hcy浓度的增加,A7r5细胞数目和吸光度(OD)值呈剂量依赖性增加,其中0.25、0.5、1 mmol/L Hcy组与对照组差异均有统计学意义(P<0.05~P<0.01)。MMP-2 mRNA的表达随着Hcy浓度的增加而增加,呈剂量依赖性,各实验组与对照组差异均有统计学意义(P<0.05)。Hcy呈剂量依赖性上调A7r5细胞MMP-2蛋白的表达,各实验组与对照组差异均有统计学意义(P<0.05~P<0.01)。结论:Hcy在一定范围内呈剂量依赖性促进A7r5细胞增殖,可在蛋白及mRNA水平使A7r5细胞MMP-2表达呈剂量依赖性增加,这一效应可能是Hcy致动脉粥样硬化的分子机制之一。

       

      Abstract: Objective: To observe the influence of homocysteine(Hcy) on the proliferation and expression of matrix metalloproteinase 2(MMP-2) in A7r5 cells.Methods: A7r5 cells were treated with 0.25,0.5 or 1 mmol/L Hcy for 48 hours.Cells counting and MTT were used todetect the proliferation of A7r5 cells.Semiquantitative reverse transcription-polymerase chain reaction(RT-PCR) was used to examine the production of MMP-2 mRNA.Western blot was used to evaluate the production of MMP-2 protein.Results: The number of A7r5 cells and optical density value increased in a dose-dependent manner when the concentration of Hcy rose.There was statistical significance between groups of 0.25,0.5,1 mmol/L Hcy and control group(P<0.05-P<0.01).Hcy increased the production of MMP-2 mRNA dose-dependently.There was statistical significance between each experimental group and control group(P<0.05).Hcy up-regulated the production of MMP-2 protein dose-dependently.There was statistical significance between each experimental group and control group(P<0.05-P<0.01).Conclusions: Hcy could promote the proliferation of A7r5 cells in a dose-dependent manner at some extent.Hcy could generate the production of MMP-2 dose-dependently both in protein and mRNA level.This effect might be one of the mechanisms involved in the atherosclerosis induced by Hcy.

       

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