t：白藜芦醇抑制可溶性CD40配体作用下巨噬细胞基质金属蛋白酶-9的表达

Resveratrol inhibits matrix metalloproteinases-9 expression induced by soluble CD40 ligand in macrophages

目的 探讨白藜芦醇抑制可溶性CD40配体（sCD40L）作用下对巨噬细胞基质金属蛋白酶-9（MMP-9）表达的影响.方法 佛波酯诱导人单核细胞株（THP-1）细胞分化为巨噬细胞.依次给予白藜芦醇和可溶性sCD40L孵育细胞,利用半定量反转录-聚合酶链反应（RT-PCR）、蛋白免疫印迹法、明胶酶谱法检测巨噬细胞内MMP-9和组织基质金属蛋白酶抑制因子-1（TIMP-1）基因的转录、蛋白表达和酶活性.结果 给予sCD40L刺激后巨噬细胞内MMP-9基因转录增多（1.53±0.04与0.75±0.01,P〈0.05）,蛋白分泌明显增加（244 930.8±31 268.6与192 976.8±20 223.1,P〈0.05） 白藜芦醇可抑制巨噬细胞MMP-9基因转录及蛋白分泌（P〈0.01）,降低MMP-9酶活性（P〈0.05）,升高巨噬细胞TIMP-1基因转录和蛋白分泌（P〈0.05）.结论 白藜芦醇可以抑制CD40途径活化的巨噬细胞内MMP-9的表达,调节MMP-9的活性,这可能是其抗动脉粥样硬化、稳定粥样斑块的作用机制之一.

Objective To explore the effect of resveratrol on the expression of matrix metalloproteinases-9 （MMP-9） in soluble CD40 ligand （sCD40L）-activated macrophages. Methods Human monocytic cell line THP-1 cells under an inducing of phorbol ester differentiated into macrophages. Then the macrophages were sitimulated by sCD40L independently and after a preincubation with resveratrol. The mRNA expression of MMP-9 and tissue-inhibitor of metalloproteinase-1 （TIMP-1） in macrophages were investigated by semiquantitative RT-PCR. The secretions of MMP-9 and TIMP-1 protein were measured by Western blot. The MMP-9 activity was analyzed by gelatin zymography technique. Results The expressions of MMP-9 gene（1.53±0.04 vs. 0.75±0.01,P〈0.05） and protein（244 930.8±31 268.6 vs. 192 976.8±20 223.1,P〈0.05）were higher in macrophages when stimulated by sCD40L. Resveratrol （10 μmol/L and 50 μmol/L）can inhibit the CD40L-induced gene expression and the protein secretion of MMP-9 （P〈0.01）. The activity of MMP-9 was degraded by resveratrol （P〈0.05）. Meanwhile resveratrol could increase the gene expression and protein secretion of TIMP-1 （P〈0.05）. Conclusions Resveratrol can inhibit the CD40L-activated macrophage expression of MMP-9. It may be one of its mechanisms on antiatherosclerosis and stabilization of atheromatous plaques.