溶血磷脂酸对基质金属蛋白酶-9表达及活性的影响

The Effect of Lysophosphatidie Acid on the Gene Expression and Activityof Matrix Metalloproteinases-9 in THP-1 Cell Line

目的观察溶血磷脂酸（Lysophosphatidie Acid,LPA）对人单核细胞株THP-1细胞基质金属蛋白酶-9（matrix metalloproteinases-9）表达及活性和NF-κB p65表达活性的影响,探讨MMP-9在LPA致动脉粥样硬化中的作用及机制。方法以不同浓度水平LPA（0-10μmol/L）刺激人THP-1细胞4h,RT-PCR法测定MMP-9mRNA表达,酶谱法检测MMP-9活性,westernblot检测核蛋白NF-κB p65表达变化。结果随着LPA剂量的增加,MMP-9表达及活性,核蛋白NF-κB p65表达逐渐增强,在LPA1μmol/L时达到最高点,然后逐渐减弱,LPA以剂量依赖的方式激活NF-κB,上调THP-1细胞MMP-9表达,增加MMP-9活性。结论LPA可能通过激活NF-κB,在转录水平促进THP-1细胞MMP-9mRNA表达,并增强其活性,促进粥样硬化发生和发展。

Objective To study the effect of lysophosphatidie acid（LPA） on the gene expression and activity of matrix metalioproteinases-9（MMP-9） and the activity of NF-κB p65 protein in inTHP-1cell line and to investigate the role of MMP-9 invovled in the atheroselerosis caused by LPA. Methods Human THP-1cells were stimulated with LPA at different concentrations（0- 10μmol/L）, the gene expression and activity of MMP-9was measured by RT-PCR and gelatin zymography respectively, the activity of NF-κB p65 was detected by wcsternblot according to the expression level of NF-κB p65 protein in cell nucleus. Results LPA upregulated the gene expression and activity of MMP-9, activated the NF-κB p65 in a dose-dependent manner, the peak point occurred at 1μmol/L LPA and decreased at higher LPA concentrations. Conclusion LPA may increase the expression and activity of MMP-9 at transcription level involving activation of NF-κB pathway in THP-1 cells which may contribute to the atherosclerosis process.