美洛昔康抑制正常人中性粒细胞与滑膜细胞粘附的作用机理研究(英文)

INHIBITORY EFFECT OF MELOXICAM ON HUMAN POLYMORPHONUCLEAR LEUKOCYTE ADHESION TO HUMAN SYNOVIAL CELL

目的 研究美洛昔康对正常人中性粒细胞 (polymorphonuclearleukocyte ,PMN)与滑膜细胞 (humansynovialcell ,HSC)粘附的抑制作用及其作用机理。方法 用MTT比色法检测PMN与HSC的粘附 ,分别用Cell ELISA和RT PCR法检测粘附分子ICAM 1和VCAM 1的蛋白及基因表达 ,用EMSA法检测NF κB的活性。结果 美洛昔康可显著的并以剂量依赖的方式抑制TNF α(5 0u·mL-1)和IL 1β(5 0 u·mL-1)作用 12h诱导的PMN与HSC粘附 ,其IC50 分别为 3 38× 10 -7和 3 5 6× 10 -6mol·L-1。进一步研究发现美洛昔康在 1×10 -6～ 1× 10 -5mol·L-1时还可在蛋白水平及mRNA水平抑制TNF α(5 0u·mL-1)诱导的HSC细胞ICAM 1的表达 ,但对VCAM 1蛋白及mRNA表达均未见显著影响 ;同时美洛昔康还可显著抑制 5 0u·mL-1TNF α诱导的NF κB的活化。结论 美洛昔康抑制NF κB的活化 ,进而抑制ICAM

AIM To investigate the effect of meloxicam on human polymorphonuclear leukocyte (PMN) adhesion to human synovial cell (HSC), and to explore its mechanism. METHODS MTT colorimetry was used to determine the adhesion effect of PMN to HSC. Cell ELISA and RT PCR methods were used to determine the expression of ICAM 1 and VCAM 1. Nuclear transcription factor kappa B (NF κB) was measured by electrophoretic mobility shift assay (EMSA) method. RESULTS Meloxicam was found to effectively inhibit TNF α (50 u·mL -1 for 12 h) and IL 1β (50 u·mL -1 for 12 h) induced adhesion of PMN to HSC (IC 50 3 38×10 -7 mol·L -1 and 3 56×10 -6 mol·L -1 , respectively) in a concentration dependent manner. ICAM 1 protein and mRNA expression induced by TNF α (50 u·mL -1 ) were inhibited by meloxicam at 1×10 -6 ～1×10 -5 mol·L -1 . The activation of NF κB was also inhibited by meloxicam at 1×10 -6 ～1×10 -5 mol·L -1 . CONCLUSION These results suggest that meloxicam inhibit TNF α stimulated PMN HSC adhesion and expression of ICAM 1 by suppressing the activity of NF κB.