Du Genlai, Liu Chengxing, Li Xiaona, Chen Weiyi, He Rui, Wang Xiaojun, Feng Pengfei, Lan Weiwei
1 Shanxi Key Lab of Material Strength & Structural Impact, Taiyuan University of Technology, Taiyuan 030024, China.
2 Department of Biology, Taiyuan Normal University, Taiyuan 030031, China.
Exp Biol Med (Maywood). 2016 Dec;241(18):2033-2041. doi: 10.1177/1535370216650940. Epub 2016 Jul 24.
Inflammatory molecules and matrix metalloproteinase (MMPs) have been found over-expressed in the tear film of patients with keratoconus. However, the mechanistic link between inflammatory molecules and MMPs in the pathogenesis of keratoconus remains still elusive. Therefore, we investigated the effect of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) on MMP-1 expression and used IL-6 antibody (IL-6 Ab) to examine the role of IL-6 on TNF-α mediated regulation of MMP-1 in fibroblasts of normal cornea and keratoconus. Real-time polymerase chain reaction, Enzyme-linked immunosorbent assay, and Western blot data demonstrated that MMP-1 and IL-6 were expressed in fibroblasts of normal cornea and keratoconus. Levels of MMP-1 and IL-6 were significantly higher in keratoconus than normal cornea. TNF-α treatment led to a significant increase in IL-6 levels. IL-6 treatment induced MMP-1 synthesis in normal cornea and keratoconus. TNF-α increased MMP-1 expression in a dose- and time-dependent manner and this response was completely inhibited by the IL-6 Ab. In conclusion, these results indicate that fibroblasts of keratoconus shows increased levels of IL-6 and MMP-1 gene and protein expression and IL-6 mediates the TNF-α-induced MMP-1 expression.
在圆锥角膜患者的泪膜中,炎症分子和基质金属蛋白酶(MMPs)已被发现过度表达。然而,在圆锥角膜发病机制中,炎症分子与MMPs之间的机制联系仍不清楚。因此,我们研究了肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)对MMP-1表达的影响,并使用IL-6抗体(IL-6 Ab)来检测IL-6在正常角膜和成圆锥角膜成纤维细胞中对TNF-α介导的MMP-1调节作用。实时聚合酶链反应、酶联免疫吸附测定和蛋白质印迹数据表明,MMP-1和IL-6在正常角膜和成圆锥角膜成纤维细胞中表达。圆锥角膜中MMP-1和IL-6的水平显著高于正常角膜。TNF-α处理导致IL-6水平显著升高。IL-6处理诱导正常角膜和成圆锥角膜中MMP-1的合成。TNF-α以剂量和时间依赖性方式增加MMP-1表达,并且这种反应被IL-6 Ab完全抑制。总之,这些结果表明,圆锥角膜成纤维细胞中IL-6和MMP-1基因及蛋白表达水平升高,并且IL-6介导TNF-α诱导的MMP-1表达。