Thapa Dinesh, Lee Jong Suk, Park Min-A, Cho Mi-Yeon, Park Young-Joon, Choi Han Gon, Jeong Tae Cheon, Kim Jung-Ae
College of Pharmacy, Yeungnam University, Gyeongsan, 712-749, Korea.
Arch Pharm Res. 2009 Apr;32(4):593-603. doi: 10.1007/s12272-009-1416-6. Epub 2009 Apr 29.
Cell adhesion molecules play a pivotal role in chronic inflammation and pathological angiogenesis. In the present study, we investigated the inhibitory effects of clotrimazole (CLT) on tumor necrosis factor (TNF)-alpha-induced changes in adhesion molecule expression. CLT dose-dependently inhibited monocyte chemoattractant protein-1 (MCP-1), intercellular cell adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) expressions in TNF-alpha-stimulated HT29 colonic epithelial cells. This inhibitory action of CLT correlated with a significant reduction in TNF-alpha-induced adhesion of monocytes to HT29 cells, which was comparable to the inhibitory effects of anti-ICAM-1 and VCAM-1 monoclonal antibodies on monocyte-epithelial adhesion. These inhibitory actions of CLT were, at least in part, attributable to the inhibition of redox sensitive NF-kappaB activation, as CLT inhibited TNF-alpha-induced ROS generation as well as NF-kappaB nuclear translocation and activation in HT29 cells. Furthermore, the inhibition of TNF-alpha-induced monocyte adhesion was also mimicked by the specific NF-kappaB inhibitor, pyrrolidine dithiocarbamate (PDTC). Inflammatory mediators including TNF-alpha have known to promote angiogenesis, which in turn further contributes to inflammatory pathology. Therefore, we additionally evaluated whether CLT modulates TNF-alpha-induced angiogenesis using in vivo chick chorioallantoic membrane (CAM) assay. The CAM assay showed that CLT dose-dependently attenuated TNF-alpha-induced angiogenesis, and the effect was correlated with decreased inflammation of the CAM tissue. In conclusion, our results suggest that CLT can inhibit TNF-alpha-triggered expression of adhesion molecules, ICAM-1 and VCAM-1, and angiogenesis during inflammation.
细胞黏附分子在慢性炎症和病理性血管生成中起关键作用。在本研究中，我们研究了克霉唑（CLT）对肿瘤坏死因子（TNF）-α诱导的黏附分子表达变化的抑制作用。CLT以剂量依赖的方式抑制TNF-α刺激的HT29结肠上皮细胞中单核细胞趋化蛋白-1（MCP-1）、细胞间细胞黏附分子-1（ICAM-1）和血管细胞黏附分子-1（VCAM-1）的表达。CLT的这种抑制作用与TNF-α诱导的单核细胞与HT29细胞黏附的显著减少相关，这与抗ICAM-1和VCAM-1单克隆抗体对单核细胞-上皮细胞黏附的抑制作用相当。CLT的这些抑制作用至少部分归因于对氧化还原敏感的NF-κB激活的抑制，因为CLT抑制了TNF-α诱导的HT29细胞中活性氧（ROS）的产生以及NF-κB的核转位和激活。此外，特异性NF-κB抑制剂吡咯烷二硫代氨基甲酸盐（PDTC）也模拟了对TNF-α诱导的单核细胞黏附的抑制作用。已知包括TNF-α在内的炎症介质可促进血管生成，而血管生成反过来又进一步促进炎症病理过程。因此，我们还使用体内鸡胚绒毛尿囊膜（CAM）试验评估了CLT是否调节TNF-α诱导的血管生成。CAM试验表明，CLT以剂量依赖的方式减弱了TNF-α诱导的血管生成，并且该作用与CAM组织炎症的减轻相关。总之，我们的结果表明，CLT可以抑制TNF-α触发的黏附分子ICAM-1和VCAM-1的表达以及炎症期间的血管生成。
