Sanggenon C对人多形核白细胞和滑膜细胞粘附的抑制作用及其机制

目的:观察化合物Sanggenon C对人外周血多形核白细胞(PMN)与人滑膜细胞(HSC)粘附的抑制作用,并探讨其作用机制.方法:MTT比色法研究PMN与HSC粘附,Cell-ELISA及RT-PCR法研究HSC粘附分子ICAM-1和VCAM-1表达,EMSA研究核转录因子NF-κB的活化.结果:Sanggenon C在0.01-10μmol·L~(-1)范围内均可显著抑制TNF-α 5O kU·L~(-1)与IL-1β诱导的HSC与PMN粘附,其IC_(50)分别为27.29nmol. L~(-1)和54.45nmol·L~(-1);Sanggenon C可显著抑制HSC表面ICAM-1和VCAM-1蛋白表达,同时也显著抑制VCAM-1 mRNA表达,但对ICAM-1 mRNA表达无显著影响;Sanggenon C在1-10μmol·L~(-1)浓度下也可显著抑制TNF-α对NF-κB的活化.结论:Sanggenon C是一个有效的人PMN与HSC粘附抑制剂,其作用机制可能是通过抑制NF-κB的活化,进而抑制HSC表面VCAM-1的表达或抑制ICAM-1转录后调控过程而实现的.

Inhibitory effect and mechanism of action of sanggenon C on human polymorphonuclear leukocyte adhesion to human synovial cells

AIM: To examine the effect of sanggenon C on human polymorphonuclear leukocyte (PMN) adhesion to human synovial cell(HSC), and explore its mechanism. METHODS: Adhesion of PMN to HSC was measured by MTT colorimetry. Cell-ELISA and RT-PCR methods were used to examine the expression of adhesion molecules ICAM-1 and VCAM-1. Activation of nuclear factor-kappa B(NF-kappaB) was measured by electrophoretic mobility shift assays(EMSA) method. RESULTS: Sanggenon C effectively inhibited TNF-alpha (50 kU/L for 12 h) and IL-1beta (50 kU/L for 12 h) induced adhesion of PMN to HSC (IC50 27.29 nmol/L and 54.45 nmol/L, respectively) in a concentration-dependent manner. Adhesion molecule VCAM-1 surface protein and mRNA expression induced by TNF-alpha 50 kU/L were significantly inhibited by sanggenon C, nevertheless, for ICAM-1 only surface protein expression being inhibited. The activation of NF-kappaB was also extensively inhibited by sanggenon C. CONCLUSION: Sanggenon C inhibited TNF-alpha -stimulated PMN-HSC adhesion and expression of VCAM-1 by suppressing the activation of NF-kappaB.