Regulation of matrix metalloprotcase-9 production induced by monocyte-vascular endothelial cell interaction - role of hepatocyte growth factor and vascular endothelial growth.

基质金属蛋白酶（MMP）已被证明在斑块破裂中起重要作用；然而，调控MMP产生的机制仍有待确定。在本研究中，我们利用人单核细胞衍生细胞系（THP-1）和人脐静脉内皮细胞（HUVEC）研究了单核细胞-血管内皮细胞（EC）相互作用对MMP产生的影响。验证肝细胞生长因子（HGF）可能调节MMP产生的假设。将THP-1与HUVEC共培养或单独在无血清培养基中培养16 h。ELISA检测培养基中MMP-2、MMP-9、HGF和血管内皮生长因子（VEGF）的生成。阻断实验采用抗hgf中和抗体（HGFAb）和抗vegf中和抗体（VEGFAb）。与单独培养THP-1相比，共培养THP-1中MMP-9的产量显著增加（92.6±4.4 vs 28.4±2.9 ng/ml, n=6; p<0.05），而MMP-2的产量不变。与单独培养THP-1相比，共培养THP-1组HGF和VEGF的生成也增加（分别为252.0±51.6 vs 182.9±29.5 pg/ml, 277.6±98,9 vs 29.5+4.8 pg/ml, n=6; p<0.05）。HGFAb显著提高THP-1共培养中MMP-9的产量，提高了1.7倍，而VEGFAb和对照IgG没有影响。这些数据建立了HGF在单核细胞- ec相互作用中抑制MMP-9产生的新功能，表明HGF在斑块稳定中起关键作用。

Matrix metalloproteases (MMP) have been shown to play an important role in plaque rupture ; however, the mechanisms that regulate MMP production remain to be determined. In the present study we investigated the effect of monocyte-vascular endothelial cell (EC) interaction on MMP production using human monocyte derived cell line (THP-1) and human umbilical vein endolhelial cells (HUVEC), and. Tested the hypothesis that hepatocyte growth factor (HGF) may regulate MMP production. THP-1 were cocultured with HUVEC or cultured alone in serum-free medium for 16 h. Production of MMP-2, MMP-9, HGF and vascular endothelial growth factor (VEGF) in the medium was examined by ELISA. Anti-HGF neutralizing antibody (HGFAb) and anti-VEGF neutralizing antibody (VEGFAb) were used for blocking studies. MMP-9 production significantly increased in cocultured THP-1 compared to THP-1 alone (92.6 ±4.4 vs 28.4 ± 2.9 ng/ml, n=6 ; p<0.05), whereas MMP-2 production was unchanged. Production of HGF and VEGF also increased in cocultured THP-1 com pared to THP-1 alone (252.0±51.6 vs 182.9±29.5 pg/ml, 277.6±98,9 vs 29.5+4.8 pg/ml, respectively, n=6 ; p<0.05). HGFAb significantly increased MMP-9 production in cocultured THP-1 by 1.7-fold, whereas VEGFAb and control IgG had no effect. These data establish a novel function of HGF in monocyte-EC interaction that inhibits MMP-9 production, indicating a critical role of HGF for plaque stabilization.