Effects of culture conditions and exposure to catabolic stimulators (IL-1 and retinoic acid) on the expression of matrix metalloproteinases (MMPs) and disintegrin metalloproteinases (ADAMs) by articular cartilage chondrocytes
Effects of culture conditions and exposure to catabolic stimulators (IL-1 and retinoic acid) on the expression of matrix metalloproteinases (MMPs) and disintegrin metalloproteinases (ADAMs) by articular cartilage chondrocytes
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DOI：
10.1016/s0945-053x(99)00024-4
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发表时间：
1999-06-01
期刊：
影响因子：
6.9
通讯作者：
Hughes, CE
中科院分区：
文献类型：
--
作者：
Flannery, CR;Little, CB;Hughes, CE
关节软骨的软骨细胞合成许多蛋白酶，这些蛋白酶能够降解这种特化细胞外基质的组分分子。类特异性蛋白酶抑制剂的使用表明，负责蛋白聚糖（聚集蛋白聚糖）和胶原蛋白的catalysts的主要活动可归因于锌依赖性金属蛋白酶。在这项研究中，我们比较了两种基质金属蛋白酶（MMP-3和MMP-13）和五种去整合素-金属蛋白酶（ADAM-10，ADAM-9，ADAM-15，TNF-α-转化酶和decysin）的软骨细胞（人，猪和牛）从新鲜软骨和软骨外植体培养和单层或琼脂糖凝胶中培养的分离细胞的mRNA表达谱。在存在或不存在白细胞介素-1（IL-1）或全反式视黄酸（两种促进体外软骨基质降解的试剂）的情况下维持此类培养物。而转录的所有金属蛋白酶检查中检测到的软骨细胞从人骨关节炎软骨在单层培养物中，ADAM-15和decysin的mRNA不存在于新鲜的骨关节炎的人软骨或外植体培养物。同样，猪和牛的金属蛋白酶mRNA的表达随不同的培养条件而变化。获得的猪和牛MMP-3和MMP-13、猪ADAM-10、猪和牛ADAM-9和猪TACE的新cDNA序列证实了关节软骨细胞表达这些分子的mRNA。定量RT-PCR分析用于确定IL-1和视黄酸对单层培养的人软骨细胞和琼脂糖培养的猪软骨细胞中金属蛋白酶mRNA水平的影响。对于MMP，IL-1处理导致人和猪MMP-3和MMP-13 mRNA增加约2 - 3倍，而视黄酸处理导致人MMP-3 mRNA水平统计学显著增加，但猪MMP-3和人或猪MMP-13的转录水平无显著变化。在暴露于IL-1或视黄酸的人类单层软骨细胞中，ADAM-15的mRNA水平升高，而TNF-α转化酶的转录物水平响应于视黄酸而增加。与此相反，ADAM-9的mRNA水平在人单层软骨细胞暴露于IL-1或视黄酸降低。结果表明，软骨细胞金属蛋白酶的表达可以在原位和体外依赖于细胞环境，并提供新的信息软骨细胞MMP和ADAM基因表达后细胞因子（IL-1）或类维生素A刺激。(C)1999年Elsevier Science B.V./国际矩阵生物学学会。All rights reserved.
The chondrocytes of articular cartilage synthesize a number of proteinases which are capable of degrading the component molecules of this specialized extracellular matrix. The use of class-specific proteinase inhibitors indicates that major activities responsible for catabolism of proteoglycan (aggrecan) and collagen are attributable to zinc-dependent metalloproteinases. In this study, we have compared the mRNA expression profiles of two matrix metalloproteinases (MMP-3 and MMP-13) and five disintegrin-metalloproteinases (ADAM-10, ADAM-9, ADAM-15, TNF-a-converting enzyme and decysin) by chondrocytes (human, porcine and bovine) from fresh cartilage and in cartilage explant cultures and isolated cells cultured in monolayer or in agarose gels. Such cultures were maintained in the presence or absence of interleukin-1 (IL-1) or all-trans-retinoic acid, two agents which promote cartilage matrix degradation in vitro. Whereas transcripts for all metalloproteinases examined were detected in chondrocytes from human osteoarthritic cartilage in monolayer cultures, mRNAs for ADAM-15 and decysin were not present in fresh osteoarthritic human cartilage or explant cultures. Similarly, expression of porcine and bovine metalloproteinase mRNAs varied with different culture conditions. Novel cDNA sequences obtained for porcine and bovine MMP-3 and MMP-13, porcine ADAM-10, porcine and bovine ADAM-9 and porcine TACE confirmed expression of mRNAs for these molecules by articular chondrocytes, Quantitative RT-PCR analysis was used to determine the effects of IL-1 and retinoic acid on metalloproteinase mRNA levels in human chondrocytes cultured in monolayer and in porcine chondrocytes cultured in agarose. For the MMPs, IL-1 treatment resulted in an approximately two to threefold increase in human and porcine MMP-3 and MMP-13 mRNAs, while retinoic acid treatment caused a statistically significant increase in human MMP-3 mRNA levels, but no significant change in transcript levels for porcine MMP-3 nor human or porcine MMP-13. The mRNA levels for ADAM-15 were elevated in hum;ln monolayer chondrocytes exposed to IL-1 or retinoic acid, while transcripts levels for TNF-alpha converting enzyme were increased in response to retinoic acid. In contrast, ADAM-9 mRNA levels were decreased in human monolayer chondrocytes exposed to IL-1 or retinoic acid. The results demonstrate that chondrocyte metalloproteinase expression can vary dependent on cell environment in situ and in vitro, and provide new information on chondrocyte MMP and ADAM gene expression following cytokine (IL-1) or retinoid stimulation. (C) 1999 Elsevier Science B.V./International Society of Matrix Biology. All rights reserved.