中国中药杂志2025，Vol.50Issue(15)：4325-4332,8.DOI:10.19540/j.cnki.cjcmm.20250409.403
基于Agrin/LRP4/MuSK信号通路探讨健脾补肾益气方促进AChR聚集改善EAMG小鼠神经-肌肉接头功能的机制研究
Mechanism of Jianpi Bushen Yiqi Decoction in promoting AChR clustering and improving neuromuscular junction function in EAMG mice based on Agrin/LRP4/MuSK signaling pathway
摘要
Abstract
This study investigated the mechanism by which Jianpi Bushen Yiqi Decoction promotes acetylcholine receptor(AChR)clustering in myasthenia gravis through the Agrin/low-density lipoprotein receptor-related protein 4(LRP4)/muscle-specific receptor tyrosine kinases(MuSK)signaling pathway.A total of 114 female C57BL/6J mice were divided into the normal group,modeling group,and solvent control group.The normal group and the solvent control group were immunized with phosphate-buffered saline(PBS),while the modeling group was established as an experimental autoimmune myasthenia gravis(EAMG)model using the murine-derived AChR-α subunit R97-116 peptide fragment.After successful modeling,the mice were randomly assigned to the model group,the low-,medium-,and high-dose Jianpi Bushen Yiqi Decoction groups,and the prednisone group.After four weeks of continuous treatment,muscle strength was assessed using Lennon scores and grip strength tests.Immunofluorescence staining was conducted on differentiated C2C12 myotubes incubated with a drug-containing serum to observe the number of AChR clusters.The integrity of AChR on myofilaments in mouse gastrocnemius muscles was further assessed by immunofluorescence staining.Hematoxylin-Eosin(HE)staining was applied to examine pathological changes in the gastrocnemius muscles of EAMG mice treated with Jianpi Bushen Yiqi Decoction.Western blot was utilized to detect the expression of key proteins in the Agrin/LRP4/MuSK signaling pathway in both C2C12 myotubes and mouse gastrocnemius muscles.The results demonstrated that compared to the model group,the prednisone group exhibited a significant decrease in the body weights of mice,whereas no significant differences in the body weights of mice were observed among the low-,medium-,and high-dose Jianpi Bushen Yiqi Decoction groups.All treatment groups showed significantly improved grip strength and Lennon scores.Additionally,the formula promoted AChR clustering on myotubes and enhanced AChR integrity in gastrocnemius myofilaments and reduced inflammatory infiltration between muscle tissue and fibrous hyperplasia.Furthermore,Jianpi Bushen Yiqi Decoction upregulated the protein expression of AChRα1,Agrin,and p-MuSK in C2C12 myotubes and increased the protein expression of AChRα1,Agrin,MuSK,p-MuSK,LRP4,and docking protein 7(Dok-7)in gastrocnemius tissue.In conclusion,Jianpi Bushen Yiqi Decoction may promote AChR clustering by targeting key proteins in the Agrin/LRP4/MuSK signaling pathway,thereby improving neuromuscular junction function and enhancing muscle strength.关键词
重症肌无力/健脾补肾益气方/Agrin/LRP4/MuSK信号通路/AChR聚集/神经-肌肉接头Key words
myasthenia gravis/Jianpi Bushen Yiqi Decoction/Agrin/LRP4/MuSK signaling pathway/AChR cluster/neuromuscu-lar junction引用本文复制引用
王家慧,刘如歌,刘韩彬,卫佳豪,张杰,柳雪影,高峰,杨俊红..基于Agrin/LRP4/MuSK信号通路探讨健脾补肾益气方促进AChR聚集改善EAMG小鼠神经-肌肉接头功能的机制研究[J].中国中药杂志,2025,50(15):4325-4332,8.基金项目
国家重大疑难疾病中西医临床协作项目(157) （157）
河南省教育厅科技攻关项目(232102311196) （232102311196）
河南中医药大学"双一流"创建工程中医学学科项目(HSRP-DFCTCM-2023-1-27) （HSRP-DFCTCM-2023-1-27）
