Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, P.R. China.
Int J Mol Med. 2013 May;31(5):1222-8. doi: 10.3892/ijmm.2013.1318. Epub 2013 Mar 26.
The etiology of osteoarthritis (OA) is complex and multifaceted. Osteoarthritis is a chronic and progressive disease of the joints that is characterized by the degradation of articular cartilage. A disintegrin and metalloproteinase with thrombospondin motifs-5 (ADAMTS-5) is the major aggrecanase in cartilage. The aim of this study was to evaluate the effect of ADAMTS-5 knockdown on cartilage degradation. Rat articular chondrocytes were transfected with lentivirus‑mediated ADAMTS-5 small interfering RNA (siRNA) or with empty vector control plasmid DNA (as the control). The suppression efficiency was measured using real-time polymerase chain reaction (RT-PCR) and western blot analysis. We then selected the most effective siRNA (siRNA1) and constructed the lentivirus-mediated siRNA targeting ADAMTS-5 for stable transfection. An animal model of OA was created using male Sprague-Dawley rats. OA was induced by performing anterior cruciate ligament transection (ACL-T) and partial medial meniscectomy (PM). The animals (n=80, weight 250‑300 g) received an intra‑articular injection of the empty vector control plasmid DNA or lentivirus‑mediated ADAMTS-5 siRNA1 (20 µl, 1x108 TU/ml). The progression of OA was analyzed using Osteoarthritis Research Society International (OARSI) scores. Compared with the control, ADAMTS-5 gene expression was decreased by approximately 80% by siRNA1 in a monolayer culture of chondrocytes. The intra‑articular injection of lentivirus‑mediated ADAMTS-5 siRNA1 in vivo resulted in the downregulation of ADAMTS-5 protein expression and improved OARSI scores (p<0.05). A single injection of lentivirus‑mediated ADAMTS‑5 siRNA prevented the degradation of articular cartilage. This method may provide a novel therapeutic strategy for the treatment of human OA.
骨关节炎（OA）的病因复杂且多方面。OA 是一种慢性进行性关节疾病，其特征是关节软骨降解。解整合素和金属蛋白酶与凝血酶反应蛋白 5（ADAMTS-5）是软骨中主要的聚集素酶。本研究旨在评估 ADAMTS-5 敲低对软骨降解的影响。用慢病毒介导的 ADAMTS-5 小干扰 RNA（siRNA）或空载体对照质粒 DNA（作为对照）转染大鼠关节软骨细胞。通过实时聚合酶链反应（RT-PCR）和蛋白质印迹分析测量抑制效率。然后选择最有效的 siRNA（siRNA1），并构建靶向 ADAMTS-5 的慢病毒介导 siRNA 进行稳定转染。使用雄性 Sprague-Dawley 大鼠建立 OA 动物模型。通过前交叉韧带切断术（ACL-T）和部分内侧半月板切除术（PM）诱导 OA。动物（n=80，体重 250-300g）接受空载体对照质粒 DNA 或慢病毒介导的 ADAMTS-5 siRNA1（20µl，1x108TU/ml）关节内注射。使用骨关节炎研究协会国际（OARSI）评分分析 OA 的进展。与对照组相比，siRNA1 在软骨细胞单层培养中使 ADAMTS-5 基因表达降低约 80%。体内注射慢病毒介导的 ADAMTS-5 siRNA1 导致 ADAMTS-5 蛋白表达下调和 OARSI 评分改善（p<0.05）。单次注射慢病毒介导的 ADAMTS-5 siRNA 可防止关节软骨降解。这种方法可能为治疗人类 OA 提供一种新的治疗策略。
