针刺研究2025，Vol.50Issue(3)：310-318,9.DOI:10.13702/j.1000-0607.20240617
电针调控miR-335-5p表达改善膝关节骨关节炎大鼠软骨组织损伤的机制研究
Mechanism of electroacupuncture in improving cartilage tissue injury in rats with knee osteoarthritis by regulating miR-335-5p expression
摘要
Abstract
Objective To observe the protective effect of electroacupuncture(EA)on the knee joint cartilage tissue of rats with knee osteoarthritis(KOA),so as to explore the potential mechanism of miR-335-5p involved in the treatment of EA for KOA.Methods A total of 42 adult male Sprague-Dawley(SD)rats were randomly divided into blank,model and EA groups,with 14 rats in each group.The model was established by injecting a sodium glutamate iodoacetate solution(80 μg/μL)into the right joint cavity.On the 15th day after the model establishment,rats in the EA group received EA intervention at"Yanglingquan"(GB34)and"Xuehai"(SP10)on the right side,and the needles were retained for 15 minutes.The intervention was carried out every other day for a total of two weeks.After the two-week intervention,the paw withdrawal mechanical threshold(PWMT)and paw withdrawal thermal latency(PWTL)of the rats were detected;a gait analyzer was used to detect the gait adaptability changes induced by pain;the pathological changes of the knee joint cartilage tissue were observed after hematoxylin-eosin(HE)staining;qPCR and Western blot were used to detect the expression levels of miR-335-5p,Dickkopf related protein 1(DKK-1),β-catenin,and matrix metalloproteinase 13(MMP-13)mRNAs and proteins in the cartilage tissue respectively;immunofluorescence staining was used to observe the positive fluorescence expression of β-catenin in the right knee joint cartilage tissue.Results In the model group,HE staining showed a large area of chondrocyte necrosis,with the dissolution and disappearance of cell nuclei,accompanied by a macrophages-predominant inflammatory cell infiltration;cartilage surface defects in local areas,necrosis of trabecular bone and proliferation of fibrous tissue in local areas,and new capillary formation in the proliferative areas were also observed.In the EA group,pathological manifestations such as inflammatory cell infiltration and fibrous tissue proliferation were significantly alleviated.Compared with the blank group,the PWMT,PWTL,the support duration,footprint area,and average ground pressure of the right hind paw,and the mRNA and protein expressions of DKK-1 in the right knee joint cartilage tissue were decreased(P<0.001)in the model group,while the expressions of miR-335-5p,mRNA and protein expressions of β-catenin and MMP-13,and the fluorescence intensity of β-catenin in the right knee joint cartilage tissue were increased(P<0.001,P<0.01).All the above indicators in the EA group were all reversed(P<0.001,P<0.05,P<0.01)in comparison with the model group.Conclusion EA can significantly improve the pain symptoms of KOA rats,promote the recovery of motor function,and improve the pathological morphology of the knee joint cartilage tissue.This may be related to regulating the expression of miR-355-5p and Wnt/β-catenin pathway,as well as inhibiting the release of cartilage extracellular matrix-related degrading enzymes.关键词
电针/膝关节骨关节炎/miR-335-5p/β-连环蛋白Key words
Electroacupuncture/Knee osteoarthritis/miR-335-5p/β-catenin引用本文复制引用
朱浩,颜雪华,陈帅,张改月,张豪斌,杨金生,王莹莹..电针调控miR-335-5p表达改善膝关节骨关节炎大鼠软骨组织损伤的机制研究[J].针刺研究,2025,50(3):310-318,9.基金项目
国家自然科学基金项目(No.82074563) （No.82074563）
国家中医药管理局中医药传承与创新"百千万"人才工程2022年青年岐黄学者培养项目(No.国中医药人教函[2022]256号) （No.国中医药人教函[2022]256号）
