肿瘤坏死因子对圆锥角膜成纤维细胞基质金属蛋白酶及其抑制剂表达的影响

Effects of Tumor Necrosis Factor Alpha on the Expression of Matrix Metalloproteinases and Tissue Inhibitors of Matrix Metalloproteinases in Keratoconus Fibroblasts

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 article is to study the effect of tumor necrosis factor alpha （TNF-α） on the expression of matrix metalloproteinases （MMPs） and tissue inhibitors of matrix metalloproteinases （TIMPs） in keratoconus fibroblasts in vitro. Normal cornea and keratoconus fibroblasts were extracted using enzyme digestion method and were cultured in the medium containing TNF-α （0, 10 and 100 ng/mL）. The expression of MMPs proteins in the supernatant of corneal fibroblasts and the expression of TIMPs in the normal cornea and keratoconus fibroblasts were detected by Western blot and real-time quantitative polymerase chain reaction respectively. The active form of MMP1 could be detected in the supernatant of keratoconus fibroblasts and upregulated by TNF-α. TNF-α could increase the protein expression of MMP2, MMP3, MMP9 in the supernatant of keratoconus fibroblasts and decrease the gene expression of TIMP1, TIMP2 in keratoconus fibroblasts. The increased MMPs and the decreased TIMPs can increase the degradation of the extracellular matrix. TNF-α may play an important role in the occurrence and development of keratoconus by regulating the expression of MMPs/TIMPs.