RESUMO
Sinomenine (SIN) has been shown to protect against IL-1ß-induced chondrocyte apoptosis in vitro. However, the role of SIN in the anterior cruciate ligament transection (ACLT)-induced osteoarthritis (OA) mouse model and its underlying molecular mechanisms remain unclear. In the present study, the protective effect of SIN on ACLT-induced articular cartilage degeneration and IL-1ß-induced chondrocyte apoptosis miR-223-3p/NLRP3 signaling regulation was investigated. Safranin O staining was performed to evaluate the pathological changes of articular cartilage. Chondrocyte apoptosis was measured with Annexin V-fluorescein isothiocyanate/polyimide (annexin V-FITC/PI) staining using flow cytometry. Gene and protein expression were detected by RT-qPCR and Western blotting, respectively. SIN administration markedly improved articular cartilage degradation in mice undergoing ACLT surgery. In addition, SIN treatment downregulated the levels of inflammatory cytokines and the protein expression of NLRP3 inflammasome components and upregulated the expression of miR-223-3p in OA mice and IL-1ß-stimulated chondrocytes. In vitro, we found that NLRP3 was a direct target of miR-223-3p, and overexpression of miR-223-3p blocked IL-1ß-induced apoptosis and the inflammatory response in chondrocytes. These findings indicate that miR-223-3p/NLRP3 signaling could be used as a potential target of SIN for the treatment of OA.
Assuntos
Doenças das Cartilagens/prevenção & controle , Cartilagem Articular/patologia , Inflamassomos/química , MicroRNAs/metabolismo , Morfinanos/farmacologia , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Animais , Antirreumáticos , Regulação da Expressão Gênica , Camundongos , Morfinanos/uso terapêutico , Substâncias Protetoras , Transdução de SinaisRESUMO
Maggot extract (ME) accelerates rat skin wound healing, however its effect on cell maintenance in wound tissues remains unclear. Bcell lymphoma (Bcl) 2associated athanogene (BAG)3 inhibits apoptosis and promotes autophagy by associating with Bcl2 or Beclin 1. Bcl2, the downstream effector of signal transducer and activator of transcription 3 signaling, is enhanced in MEtreated wound tissues, which may reinforce the Bcl2 antiapoptotic activity and/or cooperate with Beclin 1 to regulate autophagy during wound healing. The present study investigated expression levels of BAG3, Bcl2, Beclin 1 and light chain (LC)3 levels in rat skin wound tissues in the presence and absence of ME treatment. The results revealed frequent TUNELnegative cell death in the wound tissues in the early three days following injury, irrespective to ME treatment. TUNELpositive cells appeared in the wound tissues following 4 days of injury and 150 µg/ml ME efficiently reduced apoptotic rate and enhanced BAG3 and Bcl2 expression. Elevated Beclin 1 and LC3 levels and an increased LC3 II ratio were revealed in the MEtreated tissues during the wound healing. The results of the present study demonstrate the antiapoptotic effects of BAG3 and Bcl2 in MEpromoted wound healing. Beclin 1/LC3 mediated autophagy may be favorable in maintaining cell survival in the damaged tissues and MEupregulated BAG3 may enhance its activity.