OBJECTIVE:To investigate the effect of acupotomy,on mitophagy and the Pink1-Parkin pathway in chondrocytes from rabbits with knee osteoarthritis(KOA).METHODS:A KOA model was established via the modified Videman method...OBJECTIVE:To investigate the effect of acupotomy,on mitophagy and the Pink1-Parkin pathway in chondrocytes from rabbits with knee osteoarthritis(KOA).METHODS:A KOA model was established via the modified Videman method.Rabbits were randomly divided into a control group(CON),KOA group and KOA+acupotomy group(Acu).Rabbits in the acupotomy group were subjected to acupotomy for 4 weeks after model establishment.The behavior of the rabbits before and after intervention was recorded.Cartilage degeneration was evaluated by optical microscopy and fluorescence microscopy.The level of mitophagy was evaluated by transmission electron microscopy,immunofluorescence and enzyme-linked immunosorbent assay(ELISA).The expression of phosphatase and tensin homolog(PTEN)-induced kinase 1(Pink1)-Parkin mitophagy pathway components was evaluated by immunofluorescence,Western blotting and real-time polymerase chain reaction.RESULTS:In rabbits with KOA,joint pain,mobility disorders and cartilage degeneration were observed,the Mankin score was increased,collagen typeⅡ(Col-Ⅱ)expression was significantly decreased,mitophagy was inhibited,mitochondrial function was impaired,and factors associated with the Pink1-Parkin pathway were inhibited.Acupotomy regulated the expression of Pink1-Parkin pathway-related proteins,the mitophagy-related protein microtubule-associated protein-1 light chain-3,the translocase of the outer membrane,and the inner mitochondrial membrane 23;increased the colocalization of mitochondria and autophagosomes;promoted the removal of damaged mitochondria;restored mitochondrial adenosine-triphosphate(ATP)production;and alleviated cartilage degeneration in rabbits with KOA.CONCLUSIONS:Acupotomy played a role in alleviating KOA in rabbits by activating mitophagy in chondrocytes via the regulation of proteins that are related to the Pink1-Parkin pathway.展开更多
基金Key Project of Beijing University of Chinese Medicine:to Investigate the Effect of Acupotomy Mechanics on Oxidative Injury of Rabbit Chondrocytes with Knee Osteoarthritis Based on Mitophagy Mediated by Pink1/Parkin Signaling Pathway(2020-JYB-ZDGG-059)National Natural Science Foundation of China:to Investigate the Mechanism of Acupotomy"Regulating Tendon and Treating Bone"in the Treatment of Knee Osteoarthritis based on the Vascularization of Hypoxic Cartilage Mediated by Hypoxia-Inducible Factor 1alpha/Vascular Endothelial Growth Factor Pathway(82074523)。
文摘OBJECTIVE:To investigate the effect of acupotomy,on mitophagy and the Pink1-Parkin pathway in chondrocytes from rabbits with knee osteoarthritis(KOA).METHODS:A KOA model was established via the modified Videman method.Rabbits were randomly divided into a control group(CON),KOA group and KOA+acupotomy group(Acu).Rabbits in the acupotomy group were subjected to acupotomy for 4 weeks after model establishment.The behavior of the rabbits before and after intervention was recorded.Cartilage degeneration was evaluated by optical microscopy and fluorescence microscopy.The level of mitophagy was evaluated by transmission electron microscopy,immunofluorescence and enzyme-linked immunosorbent assay(ELISA).The expression of phosphatase and tensin homolog(PTEN)-induced kinase 1(Pink1)-Parkin mitophagy pathway components was evaluated by immunofluorescence,Western blotting and real-time polymerase chain reaction.RESULTS:In rabbits with KOA,joint pain,mobility disorders and cartilage degeneration were observed,the Mankin score was increased,collagen typeⅡ(Col-Ⅱ)expression was significantly decreased,mitophagy was inhibited,mitochondrial function was impaired,and factors associated with the Pink1-Parkin pathway were inhibited.Acupotomy regulated the expression of Pink1-Parkin pathway-related proteins,the mitophagy-related protein microtubule-associated protein-1 light chain-3,the translocase of the outer membrane,and the inner mitochondrial membrane 23;increased the colocalization of mitochondria and autophagosomes;promoted the removal of damaged mitochondria;restored mitochondrial adenosine-triphosphate(ATP)production;and alleviated cartilage degeneration in rabbits with KOA.CONCLUSIONS:Acupotomy played a role in alleviating KOA in rabbits by activating mitophagy in chondrocytes via the regulation of proteins that are related to the Pink1-Parkin pathway.