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本文主要研究肿瘤坏死因子-α(TNF-α)对圆锥角膜成纤维细胞基质金属蛋白酶(MMPs)及其抑制剂(TIMPs)表达的影响。利用酶消化法提取正常角膜和圆锥角膜成纤维细胞并对其施加不同浓度的TNF-α作用,通过蛋白质印迹法和实时荧光定量多聚核苷酸链式反应分别检测细胞上清夜中MMPs蛋白和细胞中TIMPs基因的表达。结果发现圆锥角膜成纤维细胞上清液中存在MMP1的活性形式,且TNF-α能使之表达上调;TNF-α能促进圆锥角膜成纤维细胞MMP2、MMP3、MMP9表达,但使其TIMP1、TIMP2表达降低。TNF-α可能通过调节MMPs和TIMPs的表达在圆锥角膜的发生发展中起着重要的作用。
The aim of this study was to investigate the effects of tumor necrosis factor-α (TNF-α) on the expression of matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) in keratoconus fibroblasts. The normal cornea and keratoconus fibroblasts were extracted by enzymatic digestion and applied with different concentrations of TNF-α. Western blotting and real-time fluorescent quantitative polymerase chain reaction were used to detect the expression of MMPs and TIMPs gene expression in cells. The results showed that the active form of MMP1 existed in the keratoconus fibroblast supernatant, and the expression of MMP1 was up-regulated by TNF-α. TNF-α could promote the expression of MMP2, MMP3 and MMP9 in keratoconus fibroblasts, Decreased expression. TNF-α may play an important role in the development of keratoconus by regulating the expression of MMPs and TIMPs.