title: Protective effect of lentivirus-mediated siRNA targeting ADAMTS-5 on cartilage degradation in a rat model of osteoarthritis.
abstract: 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.
authors.affiliation: Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, P.R. China.
authors.forename: Xiangyu
authors.initials: X
authors.lastname: Chu
authors.forename: Hongbo
authors.initials: H
authors.lastname: You
authors.forename: Xuefeng
authors.initials: X
authors.lastname: Yuan
authors.forename: Wenbin
authors.initials: W
authors.lastname: Zhao
authors.forename: Wenkai
authors.initials: W
authors.lastname: Li
authors.forename: Xin
authors.initials: X
authors.lastname: Guo
mesh_terms: D051722:ADAM Proteins / Q000235:genetics / Q000378:metabolism*; D000071116:ADAMTS5 Protein; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002358:Cartilage, Articular / Q000378:metabolism* / Q000473:pathology*; D048430:Cell Shape; D019902:Chondrocytes / Q000378:metabolism / Q000473:pathology; D024043:Collagen Type II / Q000378:metabolism; D004195:Disease Models, Animal; D005786:Gene Expression Regulation; D006801:Humans; D007596:Joints / Q000378:metabolism / Q000473:pathology; D016086:Lentivirus / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D010003:Osteoarthritis / Q000378:metabolism* / Q000473:pathology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D014162:Transfection
chemical_list: D024043:Collagen Type II; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051722:ADAM Proteins; D000071116:ADAMTS5 Protein; C518696:Adamts5 protein, rat