Vincenti M P, Coon C I, Brinckerhoff C E
Department of Medicine, Dartmouth Medical School, Hanover, New Hampshire 03755, USA.
Arthritis Rheum. 1998 Nov;41(11):1987-94. doi: 10.1002/1529-0131(199811)41:11<1987::AID-ART14>3.0.CO;2-8.
To determine how interleukin-1 (IL-1), through activation of collagenase 1 (matrix metalloproteinase 1 [MMP-1]) transcription in synovial fibroblasts, contributes to cartilage degradation in rheumatoid arthritis.
Primary rabbit synovial fibroblasts were transiently transfected with MMP-1 promoter/ luciferase constructs, and promoter activity in response to IL-1 was assessed. A minimal IL-1-response element was defined and used to evaluate DNA binding proteins by electrophoretic mobility shift assay and in situ ultraviolet crosslinking assay.
Transcriptional activation of the MMP-1 gene by IL-1 in rabbit synovial fibroblasts required a dorsal-like element, which was located at nucleotide (nt) -3,029, as well as an activator protein 1 site at nt -77. Importantly, an IL-1-induced DNA binding activity that was specific for the dorsal-like element contained the p50 subunit of nuclear factor kappaB (NF-kappaB).
These studies demonstrate, for the first time, a role for NF-kappaB in the induction of MMP-1, and suggest a mechanism of NF-kappaB-mediated cartilage degradation in rheumatoid arthritis.
确定白细胞介素-1（IL-1）如何通过激活滑膜成纤维细胞中的胶原酶1（基质金属蛋白酶1 [MMP-1]）转录，促进类风湿关节炎中的软骨降解。
用MMP-1启动子/荧光素酶构建体瞬时转染原代兔滑膜成纤维细胞，并评估其对IL-1的启动子活性。确定一个最小的IL-1反应元件，并通过电泳迁移率变动分析和原位紫外线交联分析来评估DNA结合蛋白。
IL-1在兔滑膜成纤维细胞中对MMP-1基因的转录激活需要一个位于核苷酸（nt）-3,029的背侧样元件，以及一个位于nt -77的激活蛋白1位点。重要的是，一种对背侧样元件具有特异性的IL-1诱导的DNA结合活性包含核因子κB（NF-κB）的p50亚基。
这些研究首次证明了NF-κB在MMP-1诱导中的作用，并提示了NF-κB介导类风湿关节炎中软骨降解的机制。
