痛风清消方通过OPG/RANKL/NF-κB信号通路调控湿热证痛风性关节炎大鼠骨代谢的机制研究OA

Objective To investigate the mechanism by which Tongfeng Qingxiao Formula(痛风清消方,TFQXF)regulates bone metabolism via the OPG/RANKL/NF-κB signaling pathway in rats with damp-heat gouty arthritis(GA).Methods Sixty SPF-grade male SD rats were randomly divided into six groups(n=10 per group):normal control group,model group,diclofenac sodium group,and low/medium/high-dose TFQXF groups.Except for the normal control group,all other groups were subjected to the induction of a damp-heat syndrome GA model,with corresponding drug interventions administered during the modeling period.Ankle joint swelling and gait scores were assessed in each group.HE,Safranin O-Fast Green(SO-FG),and tartrate-resistant acid phosphatase(TRAP)staining were performed to observe pathological changes in ankle joint tissues and quantify osteoclasts(OCs).Serum levels of inflammation-related markers(IL-1β,IL-10,NF-κB)and bone metabolism-related markers(OPG,RANKL,CTX-I)were measured by enzyme-linked immunosorbent assay(ELISA).The protein expression levels of OPG,RANK,and RANKL in ankle cartilage tissue were detected by Western blot(WB),while NF-κB p65 expression in synovial tissue was examined using immunohistochemistry(IHC).Results Compared with the normal group,rats in the model group exhibited significantly increased ankle joint swelling and gait scores(P<0.01),along with marked synovial inflammatory cell infiltration,cartilage structural damage,and a significant elevation in OC numbers(all P<0.01).Serum levels of IL-1β,NF-κB,RANKL,and CTX-I were significantly elevated(P<0.01),whereas IL-10 and OPG levels were markedly decreased(P<0.01).In addition,protein expression of RANK and RANKL in cartilage tissue and NF-κB p65 in synovial tissue were significantly upregulated(P<0.01),while OPG expression in cartilage tissue was reduced(P<0.01).Compared with the model group,high/medium-dose TFQXF significantly ameliorated ankle swelling and gait scores(P<0.05,P<0.01),attenuated synovial inflammation and cartilage destruction,and reduced OC numbers(P<0.01).Moreover,it downregulated serum IL-1β,NF-κB,RANKL,and CTX-I levels(P<0.05,P<0.01),upregulated IL-10 and OPG levels(P<0.01),and signifi-cantly restored the protein expression of OPG,RANKL,and RANK in cartilage tissue as well as NF-κB p65 in synovial tissue(P<0.01).The effects were comparable to those in the diclofenac sodium group.Conclusion TFQXF may mitigate joint inflammation and bone destruction in rats with damp-heat GA by regulating the OPG/RANKL/NF-κB signaling pathway,thereby inhibiting inflammatory responses,mitigating OC formation and differentiation,and modulating bone metabolic balance.