Kohyama Tadashi, Liu Xiangde, Zhu Yun Kui, Wen Fu-Qiang, Wang Hang Jun, Fang Qiuhong, Kobayashi Tetsu, Rennard Stephen I
Department of Respiratory Medicine, University of Tokyo, Japan.
Am J Respir Cell Mol Biol. 2002 Oct;27(4):487-94. doi: 10.1165/rcmb.4818.
Tissue destruction, resulting in emphysema, can be a consequence of several pathologic processes. The current study evaluated the effects of the phosphodiesterase (PDE)4 inhibitor, cilomilast, and other PDE inhibitors on the ability of fibroblasts to degrade extracellular matrix. Using the three-dimensional collagen gel culture system, fibroblasts (HFL-1) were cultured with tumor necrosis factor (TNF)-alpha, known to induce matrix metalloproteinase (MMP) release, and/or neutrophil elastase (NE), which can induce MMP activation. On Day 4, gels containing TNF-alpha and NE were significantly degraded (20.8 +/- 2.9% of original collagen content). Cilomilast (10 micro M) inhibited this degradation (84.4 +/- 8.4%). Amrinone, a PDE3 inhibitor, and zaprinast, a PDE5 inhibitor, had no effect. Gelatin zymography and immunoblotting revealed that fibroblasts cultured with TNF-alpha released increased amounts of latent MMP-1 and -9. The addition of NE resulted in the conversion of MMP-1 and -9 to their active forms, indicative of collagen degradation. Cilomilast inhibited the release of MMP-1 and -9, as well as conversion of MMP-1 to its active form. Using real-time PCR analysis, cilomilast's effect on MMP-1 release was not associated with the proteinase's mRNA expression, suggesting that the inhibition of release is regulated at the post-transcriptional level. These results suggest that cilomilast may be a potentially effective therapeutic agent in diseases characterized by excessive tissue destruction, such as emphysema.
组织破坏导致肺气肿,可能是多种病理过程的结果。本研究评估了磷酸二酯酶(PDE)4抑制剂西洛司特以及其他PDE抑制剂对成纤维细胞降解细胞外基质能力的影响。使用三维胶原凝胶培养系统,将成纤维细胞(HFL-1)与已知可诱导基质金属蛋白酶(MMP)释放的肿瘤坏死因子(TNF)-α和/或可诱导MMP激活的中性粒细胞弹性蛋白酶(NE)一起培养。在第4天,含有TNF-α和NE的凝胶被显著降解(占原始胶原含量的20.8±2.9%)。西洛司特(10μM)抑制了这种降解(84.4±8.4%)。PDE3抑制剂氨力农和PDE5抑制剂扎普司特没有效果。明胶酶谱和免疫印迹显示,与TNF-α一起培养的成纤维细胞释放出更多的潜伏MMP-1和-9。添加NE导致MMP-1和-9转化为其活性形式,表明胶原降解。西洛司特抑制了MMP-1和-9的释放以及MMP-1向其活性形式的转化。使用实时PCR分析,西洛司特对MMP-1释放的影响与该蛋白酶的mRNA表达无关,这表明释放的抑制是在转录后水平调控的。这些结果表明,西洛司特可能是一种潜在有效的治疗药物,用于治疗以组织过度破坏为特征的疾病,如肺气肿。