【摘要】 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。更多还原

【Abstract】 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 5×105·mL-1.Before A771726 treating,cells were cultured with 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.更多还原