Tsuchiya K, Maloney W J, Vu T, Hoffman A R, Huie P, Sibley R, Schurman D J, Smith R L
Orthopaedic Research Laboratory, Stanford University School of Medicine, California, USA.
J Orthop Res. 1997 Jan;15(1):94-100. doi: 10.1002/jor.1100150114.
Expression of matrix metalloproteinase-9 mRNA in osteoarthritic and normal cartilage was analyzed using reverse transcription-polymerase chain reaction and in situ hybridization. Fifty-four osteoarthritic cartilage samples were obtained from 24 patients undergoing total knee arthroplasty. Sixteen normal cartilage samples were obtained from non-osteoarthritic knees of four autopsy cases. With normal cartilage, reverse transcription-polymerase chain reaction analysis for matrix metalloproteinase-9 mRNA showed that chondrocytes exhibited only a trace signal. In analysis of osteoarthritic cartilage, chondrocytes of moderately and severely fibrillated cartilage exhibited a 73-fold and 110-fold increase in matrix metalloproteinase-9 mRNA signal, respectively, relative to normal cartilage. Chondrocytes of nonfibrillated osteoarthritic cartilage exhibited a 6-fold increase (p < 0.02) in matrix metalloproteinase-9 mRNA signal relative to normal cartilage. Analysis of matrix metalloproteinase-9 mRNA expression in fresh-frozen sections of normal and osteoarthritic cartilage by in situ hybridization confirmed these results. This study showed that reverse transcription-polymerase chain reaction provides a sensitive index of mRNA levels in normal and osteoarthritic cartilage samples and suggests that increased expression of matrix metalloproteinase-9 precedes fibrillation of cartilage in the development of osteoarthritis.
采用逆转录-聚合酶链反应和原位杂交技术分析骨关节炎软骨和正常软骨中基质金属蛋白酶-9 mRNA的表达。从24例行全膝关节置换术的患者中获取54份骨关节炎软骨样本。从4例尸检病例的非骨关节炎膝关节中获取16份正常软骨样本。对于正常软骨，基质金属蛋白酶-9 mRNA的逆转录-聚合酶链反应分析显示软骨细胞仅呈现微量信号。在骨关节炎软骨分析中，中度和重度纤维化软骨的软骨细胞中基质金属蛋白酶-9 mRNA信号相对于正常软骨分别增加了73倍和110倍。未纤维化的骨关节炎软骨的软骨细胞中基质金属蛋白酶-9 mRNA信号相对于正常软骨增加了6倍（p < 0.02）。通过原位杂交对正常和骨关节炎软骨新鲜冰冻切片中基质金属蛋白酶-9 mRNA表达的分析证实了这些结果。本研究表明，逆转录-聚合酶链反应为正常和骨关节炎软骨样本中的mRNA水平提供了一个敏感指标，并提示在骨关节炎发展过程中，基质金属蛋白酶-9表达增加先于软骨纤维化。
