杜仲-牛膝配伍抗类风湿关节炎机制的网络药理学和实验验证研究OA

Objective To explore the mechanism of the Cortex Eucommiae(CE)-Radix Achyranthis Bidentatae(RAB)medicinal cou-plet against rheumatoid arthritis(RA).Methods A collagen-induced arthritis(CIA)rat model was established.Paw swelling volume and arthritis scores were recorded after drug administration.HE staining and safranin O/fast green staining were used to observe ankle joint pathology.Serum levels of bone metabolism markers and inflammatory cytokines were detected.Scratch wound healing and Tran-swell assays were conducted to evaluate the migration and invasion capabilities of HFLS-RA cells.Tartrate-resistant acid phosphatase(TRAP)staining was employed to assess osteoclast differentiation of RAW264.7 cells.The mRNA levels of related factors were deter-mined by reverse transcription-quantitative polymerase chain reaction(RT-qPCR).Results Both individual herbs(CE and RAB)and their medicinal couplet(CE-RAB)reduced the paw swelling volume and arthritis scores in CIA rats.CE and CE-RAB reduced serum levels of β-cross-linked C-telopeptide of type I collagen(β-CTX),pyridinoline cross-linked carboxy-terminal telopeptide of type I col-lagen(ICTP),nitric oxide(NO),IL-1β,and TNF-α,while increasing the levels of procollagen type I N-terminal propeptide(PINP)and bone gla protein(BGP).In spleen tissue,CE,RAB,and CE-RAB downregulated the mRNA expression of IL-6,IL-1β,and TNF-α.Additionally,RAB and CE-RAB upregulated the mRNA expression of IL-10.CE,RAB,and CE-RAB inhibited the migration of HFLS-RA cells and downregulated the mRNA expression of IL-6,IL-1β,and TNF-α in these cells.Furthermore,CE and CE-RAB also inhibited cell invasion.CE,RAB,and CE-RAB suppressed the differentiation of RAW264.7 cells into osteoclasts.Moreover,CE and CE-RAB reduced the mRNA expression of IL-6,IL-1β,and TNF-α in RAW264.7 cells.CE,RAB,and CE-RAB downregulated IL-17A mRNA levels in the ankle joints of CIA rats and in HFLS-RA cells.Conclusion The CE-RAB alleviated arthritis in CIA rats,mitigated bone metabolism disorder and pathological damage,suppressed the migration,invasion,and inflammatory factor expression in HFLS-RA cells,and inhibited osteoclast differentiation.The synergistic mechanism likely involves the TNF and IL-17 signaling path-ways,with CE appearing to play a predominant role.