施氏滋肾通痹汤调控Wnt/β-catenin信号通路治疗膝骨关节炎的作用机制OA

Objective:To investigate the therapeutic mechanism of Shishi Zishen Tongbi decoction in treating knee osteoar-thritis(KOA)through the β-catenin signaling pathway.Methods:Thirty male SPF-grade Sprague-Dawley rats were ran-domly divided into three groups:the sham-operated group,the model group,and the Shishi Zishen Tongbi decoction group(n=10 each).A modified Hulth method was used to establish KOA models.After two months of modeling,the Shishi Zishen Tongbi decoction group received intragastric administration of the decoction for an additional two months.Upon completion,knee joints were collected for evaluation.X-ray and Micro-CT were used to assess structural changes in sub-chondral bone.Cartilage morphology was observed using ABH/OG and Safranin O/Fast Green staining.Immunohisto-chemistry was performed to detect the expression of β-catenin,Col X,MMP-13,and Col Ⅱ.RT-PCR and Western Blot were used to measure the mRNA and protein levels of Wnt3a,MMP-9,MMP-13,Col X,alkaline phosphatase(ALP),osteocalcin(OC)and Axin2.Results:X-ray and Micro-CT analyses demonstrated a significant increase in osteophyte for-mation around the knee joint in the model group compared with the sham-operated group.However,osteophyte formation was markedly reduced in the Shishi Zishen Tongbi decoction group compared to the model group.Histological evaluations using ABH/OG and Safranin O/Fast Green staining showed a smooth articular cartilage surface and clearly defined four-layer structures(superficial,transitional,radial,and calcified zones)in the sham-operated group,with chondrocytes arranged in regular columnar patterns and no evidence of chondrocyte clustering.In contrast,the model group exhibited re-duced cell numbers in the superficial zone,prominent chondrocyte clustering in the transitional and radial zones,forward displacement of the tidemark,and double tidemark formation.The cartilage morphology in the Shishi Zishen Tongbi decoction group was significantly improved relative to that in the model group.Immunohistochemical analysis revealed sig-nificantly elevated expression levels of β-catenin,Col X,and MMP-13 in the cartilage tissue of the model group compared to the sham-operated group(P<0.05),while these protein levels were significantly reduced in the Shishi Zishen Tongbi decoction group compared to the model group(P<0.05).Real-time PCR results further confirmed that the expression levels of Wnt3a,MMP9,MMP13,Col X,ALP,OC and Axin2 genes were significantly higher in the model group than in the sham-operated group(P<0.05),whereas these gene expression levels were significantly downregulated in the Shishi Zishen Tongbi decoction group compared to the model group(P<0.05).Conclusion:Shishi Zishen Tongbi decoction exerts therapeutic effects on KOA by modulating the β-catenin signaling pathway,likely through downregulation of key genes and pro-teins involved in its activation,thus slowing disease progression.