NF-κB通路在TNF-α调控软骨细胞MMPs/TIMPs比例中的作用研究

The Effects of NF-kappa B Signalling on the TNF-alpha-induced Ratios of MMPs to TIMPs in Chondrocytes

中文关键词:  软骨细胞 肿瘤坏死因子-α 核因子-κB 基质金属蛋白酶家族 基质金属蛋白酶组织抑制剂家族

英文关键词:Chondrocyte TNF-α NF-κB MMPs TIMPs

中文摘要:

      目的 探索核因子-κB（NF-κB）对肿瘤坏死因子-α（TNF-α）诱导下软骨细胞中基质金属蛋白酶家族（MMPs）与基质金属蛋白酶组织抑制剂家族（TIMPs）比值的影响。方法 采用胰酶消化法从1日龄小鼠的膝关节处提取软骨细胞。HE染色以显示软骨细胞形态；半定量PCR用于分析TNF-α诱导下bay11-7082对软骨细胞中MMPs和TIMPs基因表达的影响，酶谱用于鉴定TNF-α和bay11-7082作用后明胶酶活性的改变。结果 TNF-α能使MMPs和TIMPs的基因表达上调（P <0.05）。MMPs （除外MMP-1）与TIMPs 的比值均出现上调（P <0.05）。Bay11-7082能抑制TNF-α诱导下MMPs和TIMPs的表达上调，并能使MMPs/TIMPs 上调的比值下降到接近于正常软骨细胞的水平。酶谱检测明胶酶的变化，也有一致的发现。结论 TNF-α诱导下MMPs与TIMPs比值的上升可在一定程度上解释骨关节炎（OA）中软骨基质的过度降解。用bay11-7082阻断NF-κB途径可能为OA的治疗提供方案。

英文摘要:

      Objective To explore the influence of the NF-κB inhibitor (bay11-7082) on tumor necrosis factor-alpha (TNF-α)-induced different ratios of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in chondrocytes. Methods Chondrocytes were isolated from the knee joint of a 1-day old mouse by trypsin digestion method. Hematoxylin-Eosin (HE) stain was used to show the morphology of isolated chondrocytes; Semi-quantitative PCR was applied to analyze the influence of bay11-7082 on gene expressions of TNF-α-induced MMPsand TIMPsin chondrocytes; Zymography was used to elucidate activities of the gelatinases induced by TNF-α and/or bay11-7082. Results TNF-α up-regulated gene expressiosn of the MMPsand TIMPs(P<0.05). The ratios of MMPs/TIMPswere mostly increased except the part of MMP-1. Bay11-7082 could reduce TNF-α-induced MMPsand TIMPsgene expressions, and could make the increased ratio of MMPs/TIMPsdropped to the normal level of chondrocytes. Similar results were observed at the protein level of the gelatinases by zymography. Conclusion TNF-α-induced high ratios of MMPs/TIMPs could partiallyexplain over-degradation of cartilage extracellular matrix in osteoarthritis (OA). Blockage of NF-κB with bay11-7082 might provide a possible therapeutic strategy for the OA deterioration.