Fortier Lisa A, Schnabel Lauren V, Mohammed Hussni O, Mayr Karen G
Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
Am J Vet Res. 2007 Apr;68(4):379-84. doi: 10.2460/ajvr.68.4.379.
To determine the effects of matrix metalloproteinase (MMP)-13, compared with interleukin (IL)-1alpha, on cartilage matrix molecule gene expression in a coculture system of equine cartilage explants and synoviocytes.
Articular cartilage and synovium specimens harvested from femoropatellar joints of 4 horses, aged 3 to 5 years.
Synoviocytes were isolated and cocultured with cartilage explants. Cultures were treated with human recombinant MMP-13 (1, 25, or 100 ng/mL) or IL-1alpha (0.01, 0.1, 1.0, or 10 ng/mL) for 96 hours, with medium exchange at 48 hours. Cartilage extracts and media were analyzed for glycosaminoglycan (GAG) content, and results were adjusted to cartilage DNA content. Quantitative PCR was performed on mRNA from cartilage (MMP-3, MMP-13, aggrecan, and collagen type IIB [COL2A1]) and synoviocytes (MMP-3 and MMP-13), and results were adjusted to 18S ribosomal subunit mRNA expression. Treatments were performed in triplicate, and the experiment was repeated 4 times.
Cultures treated with MMP-13 or IL-1alpha had increased media GAG concentration at 48 and 96 hours. Aggrecan and COL2A1 mRNA expression were increased by application of MMP-13 or IL-1alpha. Gene expression of the catabolic mediator, MMP-3, in cartilage and synoviocytes was increased in cultures treated with MMP-13 or IL-1alpha. Expression of MMP-13 mRNA in cartilage was increased by IL-1alpha, but decreased in synoviocytes by MMP-13 treatment.
Results support the use of recombinant MMP-13 in a coculture system of synoviocytes and cartilage explants for the study of osteoarthritis.
在马软骨外植体与滑膜细胞的共培养系统中，确定基质金属蛋白酶（MMP）-13 与白细胞介素（IL）-1α相比，对软骨基质分子基因表达的影响。
从 4 匹 3 至 5 岁马的股髌关节采集的关节软骨和滑膜标本。
分离滑膜细胞并与软骨外植体共培养。培养物用重组人 MMP-13（1、25 或 100 ng/mL）或 IL-1α（0.01、0.1、1.0 或 10 ng/mL）处理 96 小时，48 小时时更换培养基。分析软骨提取物和培养基中的糖胺聚糖（GAG）含量，并将结果调整为软骨 DNA 含量。对软骨（MMP-3、MMP-13、聚集蛋白聚糖和 IIB 型胶原[COL2A1]）和滑膜细胞（MMP-3 和 MMP-13）的 mRNA 进行定量 PCR，结果调整为 18S 核糖体亚基 mRNA 表达水平。处理重复 3 次，实验重复 4 次。
用 MMP-13 或 IL-1α处理的培养物在 48 和 96 小时时培养基中 GAG 浓度升高。应用 MMP-13 或 IL-1α可使聚集蛋白聚糖和 COL2A1 mRNA 表达增加。在用 MMP-13 或 IL-1α处理的培养物中，分解代谢介质 MMP-3 在软骨和滑膜细胞中的基因表达增加。IL-1α可使软骨中 MMP-13 mRNA 表达增加，但 MMP-13 处理可使滑膜细胞中 MMP-13 mRNA 表达降低。
结果支持在滑膜细胞与软骨外植体的共培养系统中使用重组 MMP-13 来研究骨关节炎。
