A771726对经PMA活化的THP-1细胞MMP-2及MMP-9表达的影响

目的 观察来氟米特活性代谢产物A771726对经PMA活化的THP-1细胞MMP-2及MMP-9表达的影响。方法 THP-1细胞悬浮生长于含10%胎牛血清的RPMI 1640培养液中。细胞密度达5×105·ml-1时用于实验。THP-1细胞经PMA刺激24h（诱导THP-1细胞分化成巨噬细胞）后，细胞进行无血清化处理12h,继而加入不同剂量的A771726（5ug/mL、15 ug/mL 、45ug/mL），干预24h。实时荧光定量PCR检测细胞MMP-2、MMP-9 mRNA的表达，明胶酶谱法检测细胞培养上清液中MMP-2、MMP-9的活性。结果 细胞经不同浓度的A771726（5ug/mL、15 ug/mL 、45ug/mL）处理后，MMP-2、MMP-9 mRNA表达量均有下降(P＜0.01) 。中、高剂量的A771726（15 ug/mL 、45ug/mL）可降低PMA活化的THP-1细胞培养上清液中MMP-2、MMP-9活性(P＜0.01)。结论 来氟米特活性代谢产物A771726可以降低经PMA活化的THP-1细胞MMP-2及MMP-9的表达。

The effects of A771726 on expression of MMP-2 and MMP-9 in PMA-activated THP-1 cells

Objective To investigate the effects of the leflunomide active metabolite (A771726) on the expression of MMP-2 and MMP-9 in PMA-actived THP-1 cells. Methods THP-1 cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum. For all experiments, THP-1 cells were cultured at an initial density of 5×105?ml-1. Before A771726 treating, cells were cultured with serum-free RPMI-1640 medium for 12h. PMA-actived THP-1 cells (THP-1 cells were induced to differentiate into macrophages with PMA for 24h)were treated with different concentrations of A771726 (5ug/ml、15 ug/ml、45ug/ml ) for 24h. The mRNA expression of MMP-2 and MMP-9 was measured by real-time PCR. The activity of MMP-2 and MMP-9 were by gelatin zymography.Results The expression of MMP-2、MMP-9 mRNA was inhibited significantly by A771726（5 ug/ml 、15 ug/ml 、45ug/ml）(P＜0.01).The acticity of MMP-2、MMP-9 in the supernatant was inhibited significantly by A771726(15 ug/ml 、45ug/ml）(P＜0.01). Conclusion Leflunomide active metabolite( A771726) can decrease the expression of MMP-2 and MMP-9 in THP-1 cells.