针刺研究2025，Vol.50Issue(10)：1114-1123,10.DOI:10.13702/j.1000-0607.20240990
电针对膝关节骨关节炎兔肠道菌群及关节软骨TLR4/MyD88/NF-κB信号通路的影响
Effects of electroacupuncture on intestinal flora and TLR4/MyD88/NF-κB signaling pathway of articular cartilage in rabbits with knee osteoarthritis
摘要
Abstract
Objective To observe the effect of electroacupuncture(EA)on knee function and symptoms,intestinal flora and TLR4/MyD88/NF-κB signaling pathway of articular cartilage in rabbits with knee osteoarthritis(KOA),so as to explore its potential mechanism underlying improvement of knee function by regulating the gut-joint axis response of KOA.Methods Thirty-two Japanese white rabbits were randomly divided into blank,model,glucosamine and EA groups,with 8 rabbits in each group.The KOA model was reproduced by using modified Hulth method.The rabbits in the EA group were given EA(2 Hz/100 Hz,0.5-1.5 mA)at bilateral"Xuehai"(SP10),"Yinlingquan"(SP9)and"Dubi"(ST35)for 15 min,once daily,5 times a week for 4 weeks.The rabbits of the glucosamine group received glucosamine solution once daily,5 times a week for 4 weeks.The knee function was evaluated by Lequesne MG index of severity for osteoarthritis(total scores of pain,swelling,range of motion,and gait)after modeling and treatment.H.E.staining was used to observe the histopathological changes of the knee cartilage tissue.Immunofluorescence histochemical method was used to detect the expressions of MyD88 and NF-κB p65,and Wes automated protein blot quantitative analysis technique was employed to detect the protein expressions of TLR4,MyD88 and NF-κB p65 in the knee cartilage tissue.Analysis of the species structure changes of intestinal flora was detected by 16S rDNA high-throughput sequencing.Results After modeling,compared with the blank group,the Lequesne MG score,immunofluorescence intensity of MyD88 and NF-κB p65,and the protein expression levels of TLR4,MyD88 and NF-κB p65 were significantly increased in the model group(P<0.05).H.E.staining showed thin and rough surface of the knee cartilage,clusters and disordered arrangement of chondrocytes,a few cellular nuclei being dark blue-purple in staining,with some nuclei being shrank,ruptured,and disappeared in the model group.In comparison with the model group,the Lequesne MG score,fluorescence intensity of MyD88 and NF-κB p65,and the protein expression levels of TLR4,MyD88 and NF-κB p65 were significantly decreased in both EA and glucosamine groups(P<0.05).The effects of EA were significantly superior to glucosamine in down-regulating the protein expression levels of TLR4,MyD88 and NF-κB(P<0.05).H.E.staining showed relatively complete surface of the knee cartilage,slightly hypertrophic chondrocytes with relatively clear nucleus and relatively complete tide line in both EA and glucosamine groups.Results of 16S rDNA sequencing showed that compared with the blank group,the Shannon index was obviously increased(P<0.05),the relative abundance of Proteobacteria,Actinobacteria,Escherichia coli-Shigella and Clostridium UCG-014 was elevated,and the relative abundance of Firmicutes,Verrucomicrobia,Comamonas and Acmea bacteria was decreased in the model group.In comparison with the model group,the Chao1 index and Shannon index were increased(P<0.05),the relative abundance of Proteobacteria,Actinobacteria and Escherichia coli-Shigella was decreased,while the relative abundance of Firmicutes,Comamonas and Nobilus was significantly increased in both EA and glucosamine groups.Conclusion EA can significantly improve the symptoms and function of knee joint in KOA rabbits,which may be associated with its functions in inhibiting cartilage injury and inflammation reaction by regulating TLR4/MyD88/NF-κB signaling pathway and intestinal flora diversity through the"gut-joint"axis.关键词
膝关节骨关节炎/电针/Toll样受体4(TLR4)/髓样分化因子88(MyD88)/核转录因子-κB(NF-κB)信号通路/炎性反应/肠道菌群Key words
Knee osteoarthritis/Electroacupuncture/TLR4/MyD88/NF-κB signaling pathway/Inflammation/Intestinal flora引用本文复制引用
胡哲,马俊杰,马铁明,王列,于嘉祥,于冬冬,王树东..电针对膝关节骨关节炎兔肠道菌群及关节软骨TLR4/MyD88/NF-κB信号通路的影响[J].针刺研究,2025,50(10):1114-1123,10.基金项目
辽宁省科技厅面上项目(No.2023-MS-229) （No.2023-MS-229）
辽宁省教育厅课题(No.LJKZ0878) （No.LJKZ0878）
