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

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

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RPM I 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24 h(诱导THP-1细胞分化成巨噬细胞)后,细胞进行无血清化处理12 h,继而加入不同剂量的A771726(5、15、45μg/mL),干预24 h。实时荧光定量PCR检测细胞MMP-2、MMP-9mRNA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、45μg/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。

Objective To investigate the effects of leflunomide active metabolite （A771726） on the expression of MMP -2 and MMP -9 in PMA - activated 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 started with an initial density of 5 × 10^5 · mL^-1. Before A771726 treating, cells were cultured wifll serum - free RPMI - 1640 medium for 12h. PMA - activated THP - 1 cells （ THP - 1 cells were induced to differentiate into macrophages with PMA for 24 h ） were treated with different concentrations of A771726 （5, 15 and 45 μg/mL） for 24h. The mRNA expression and activity of MMP -2 and MMP - 9 was qualified by real - time PCR and gelatin zymography, respectively. Results The expression of MMP - 2 and MMP -9 mRNA was significantly inhibited by A771726 （5, 15 and 45 μg/mL, P 〈0. 01 ）. The activity of MMP - 2 and MMP- 9 was inhibited by A771726（ 15 and 45ug/mL）in the supernatant of PMA- activated THP- 1 cells （P 〈 0. 01 ）. Conclusion Leflunomide active metabolite （A771726） can decrease the expression of MMP -2 and MMP -9 in PMA - activated THP - 1 cells.