Objective:To explore the effect of acupotomy intervention on autophagy of chondrocytes in rabbits with knee osteoarthritis(KOA),and to determine the possible mechanisms of acupotomy to alleviate cartilage degeneration...Objective:To explore the effect of acupotomy intervention on autophagy of chondrocytes in rabbits with knee osteoarthritis(KOA),and to determine the possible mechanisms of acupotomy to alleviate cartilage degeneration.Methods:The modified Videman method was used to construct a KOA rabbit model.After modeling,40 rabbits were randomly divided into 4 groups by a random number table:control;KOA(model);KOA+acupotomy(acupotomy),and KOA+sham acupotomy(sham),10 in each group.After a 3-week treatment course,the knee joint activity was determined by the modified Lequesne MG index.Hematoxylin-eosin staining staining was used to examine the morphological changes of chondrocytes.Autophagy of chondrocytes was observed by transmission electron microscopy.The surface morphology of cartilage tissue was observed by scanning electron microscope.The mRNA and protein levels of AMP kinase/mammalian target of rapamycin/Unc-51(AMPK/mTOR/ULK1)signal pathway key proteins,autophagy-related factor Beclin-1 and microtubule-associated protein 1A/1B light chain 3(LC3)in rabbit knee cartilage were assessed by real-time fluorescence quantitative polymerase chain reaction and Western blot,respectively.Results:The modified Lequesne MG score of acupotomy group was significantly lower than that of model group(P<0.05).Pathological results showed that chondrocyte autophagy decreased and cartilage surface was rough in the model group,which recovered after acupotomy treatment.The mRNA expressions of AMPK,ULK1,Beclin-1 and the protein levels of p-AMPK,p-ULK1,Beclin-1,and LC3Ⅱ/LC3Ⅰwere decreased in the model group,while the mRNA and protein expressions of mTOR were increased(P<0.01).However,acupotomy treatment reversed these abnormal changes(P<0.05).Conclusions:Acupotomy could effectively up-regulate the expressions of AMPK,ULK1 and Beclin1,reduce the expression of mTOR,promote autophagy,and alleviate joint degeneration.Acupotomy is a promising complementary and alternative therapy for KOA.展开更多
基金Supported by the National Natural Science Foundation of China(No.81774436)the Science and Technology Innovation Fund Project of Anhui University of Traditional Chinese Medicine(No.2021ZC02)the“14th Five-Year Plan”Provincial Traditional Chinese Medicine Advantage Characteristic Specialty Project[No.(2021]71)]。
文摘Objective:To explore the effect of acupotomy intervention on autophagy of chondrocytes in rabbits with knee osteoarthritis(KOA),and to determine the possible mechanisms of acupotomy to alleviate cartilage degeneration.Methods:The modified Videman method was used to construct a KOA rabbit model.After modeling,40 rabbits were randomly divided into 4 groups by a random number table:control;KOA(model);KOA+acupotomy(acupotomy),and KOA+sham acupotomy(sham),10 in each group.After a 3-week treatment course,the knee joint activity was determined by the modified Lequesne MG index.Hematoxylin-eosin staining staining was used to examine the morphological changes of chondrocytes.Autophagy of chondrocytes was observed by transmission electron microscopy.The surface morphology of cartilage tissue was observed by scanning electron microscope.The mRNA and protein levels of AMP kinase/mammalian target of rapamycin/Unc-51(AMPK/mTOR/ULK1)signal pathway key proteins,autophagy-related factor Beclin-1 and microtubule-associated protein 1A/1B light chain 3(LC3)in rabbit knee cartilage were assessed by real-time fluorescence quantitative polymerase chain reaction and Western blot,respectively.Results:The modified Lequesne MG score of acupotomy group was significantly lower than that of model group(P<0.05).Pathological results showed that chondrocyte autophagy decreased and cartilage surface was rough in the model group,which recovered after acupotomy treatment.The mRNA expressions of AMPK,ULK1,Beclin-1 and the protein levels of p-AMPK,p-ULK1,Beclin-1,and LC3Ⅱ/LC3Ⅰwere decreased in the model group,while the mRNA and protein expressions of mTOR were increased(P<0.01).However,acupotomy treatment reversed these abnormal changes(P<0.05).Conclusions:Acupotomy could effectively up-regulate the expressions of AMPK,ULK1 and Beclin1,reduce the expression of mTOR,promote autophagy,and alleviate joint degeneration.Acupotomy is a promising complementary and alternative therapy for KOA.
