title: Inhibition of microRNA-449a prevents IL-1β-induced cartilage destruction via SIRT1.
abstract: SIRT1 has anti-inflammatory as well as protective effects in chondrocytes. The object of this study was to investigate whether microRNA-449a regulates expression of SIRT1, which inhibits expression of catabolic genes in IL-1β-induced cartilage destruction.
  
  
  
  MicroRNA-449a expression was determined in OA chondrocytes and IL-1β-induced chondrocytes by real-time PCR. MicroRNA-449a binding sites on the 3'-UTR of SIRT1 mRNA and binding site conservation were examined using microRNA target prediction tools. SIRT1-overexpressing or knockdown chondrocytes were transfected with microRNA-449a or anti-microRNA-449a mimic and stimulated by IL-1β. Expression of catabolic and anabolic genes was examined by real-time PCR and western blotting. Finally, positive effects of anti-microRNA-449a on expression of these genes were confirmed by western analysis of OA chondrocytes.
  
  
  
  Expression of microRNA-449a was increased in OA chondrocytes and IL-1β-induced chondrocytes. MMP-13 expression was enhanced, whereas type II collagen and SIRT1 expression were decreased in IL-1β-induced chondrocytes. SIRT1 overexpression resulted in decreased expression of catabolic genes such as MMPs and ADAMTSs in response to IL-1β, but these effects were moderated by microRNA-449a. Suppression of microRNA-449a by anti-microRNA-449a inhibited expression of catabolic genes despite IL-1β stimulation, but these effects were abolished in SIRT1 knockdown chondrocytes. Furthermore, expression of catabolic genes was decreased and expression of type II collagen as well as SIRT1 was restored by anti-microRNA-449a in OA chondrocytes as well as in IL-1β-induced chondrocytes.
  
  
  
  Silencing of microRNA-449a had a protective effect, inhibiting catabolic gene expression and restoring anabolic gene expression, by targeting SIRT1 in IL-1β-induced cartilage destruction.
authors.affiliation: Department of Orthopedic Surgery, Severance Hospital, Yonsei University College of Medicine, Seoul, South Korea; Brain Korea 21 Project Plus for Medical Science, Yonsei University College of Medicine, Seoul, South Korea.
authors.forename: K W
authors.initials: KW
authors.lastname: Park
authors.affiliation: Department of Orthopedic Surgery, Severance Hospital, Yonsei University College of Medicine, Seoul, South Korea; Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, South Korea.
authors.forename: K-M
authors.initials: KM
authors.lastname: Lee
authors.affiliation: Department of Orthopedic Surgery, Severance Hospital, Yonsei University College of Medicine, Seoul, South Korea.
authors.forename: D S
authors.initials: DS
authors.lastname: Yoon
authors.affiliation: Department of Orthopaedic Surgery, CHA Bundang Medical Center, CHA University, Kyeonggi-do, South Korea.
authors.forename: K H
authors.initials: KH
authors.lastname: Park
authors.affiliation: Department of Orthopedic Surgery, Severance Hospital, Yonsei University College of Medicine, Seoul, South Korea.
authors.forename: W J
authors.initials: WJ
authors.lastname: Choi
authors.affiliation: Department of Orthopedic Surgery, Severance Hospital, Yonsei University College of Medicine, Seoul, South Korea; Brain Korea 21 Project Plus for Medical Science, Yonsei University College of Medicine, Seoul, South Korea; Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, South Korea.
authors.forename: J W
authors.initials: JW
authors.lastname: Lee
authors.affiliation: Department of Orthopedic Surgery, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, South Korea. Electronic address: orthohwan@gmail.com.
authors.forename: S-H
authors.initials: SH
authors.lastname: Kim
mesh_terms: D002356:Cartilage; D002478:Cells, Cultured; D019902:Chondrocytes; D006801:Humans; D053583:Interleukin-1beta; D035683:MicroRNAs
chemical_list: D053583:Interleukin-1beta; D035683:MicroRNAs
keywords: Chondrocytes; IL-1β; Osteoarthritis; SIRT1; microRNA-449a