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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 1的表达可能是其抑制PMN与HSC粘附的机制之一
Objective To study the inhibitory effect of meloxicam on the adhesion of polymorphonuclear leukocyte (PMN) and synovial cells (HSC) to normal human and its mechanism. Methods MTT assay was used to detect the adhesion of PMN to HSC. The protein and gene expression of adhesion molecules ICAM 1 and VCAM 1 were detected by Cell ELISA and RT PCR respectively. The activity of NF κB was detected by EMSA. Results Meloxicam significantly and in a dose-dependent manner inhibited the adhesion of PMN to HSC induced by TNFα (50μ · mL-1) and IL-1β (50μ · mL-1) 3 38 × 10 -7 and 3 5 6 × 10 -6 mol·L -1. Further study found that meloxicam at 1 × 10 -6 ~ 1 × 10 -5 mol·L-1 also inhibits TNFα (50 u · mL-1) -induced HSC ICAM 1 protein level and mRNA levels But no significant effect on the expression of VCAM1 protein and mRNA. Meanwhile, meloxicam also significantly inhibited the activation of NFκB induced by 50u · mL-1 TNFα. Conclusions Meloxicam inhibits the activation of NF-κB, and then inhibits the expression of ICAM-1, which may be one of the mechanisms by which meloxicam suppresses the adhesion of PMN to HSC