Jiawei Yanghe Decoction Ameliorates Cartilage Degradation In OA Rats Via Wnt/?-catenin Signaling Pathway

Objective:To investigate the effect of Jiawei Yanghe decoction(JWYHD)on the OA rat model and to observe its mechanism based on Wnt/ ?-catenin signaling pathway.Method:IL-1 ? was used to induce rat articular chondrocytes cultured in vitro and treated with JWYHD at gradient concentration.MTT and DAPI staining were used to detect the effect of JWYHD on the proliferation rate of cultured rat articular chondrocytes.Western Blot and qRT-PCR were used to detect the effect of JWYHD on the expression of ?-catenin,CyclinD1,MMP-3,MMP-13,Caspase-3,Caspase-9 and Col2a1.The effect of JWYHD on extracellular matrix of chondrocytes was detected by Alcian blue staining.Alternatively,DKK-1 and Licl were used to observe the effect of JWYHD on Wnt/ ?-catenin signaling pathway.50 SD rats were randomly divided into blank,model,low,middle and high dose groups.The OA rat model was induced by injection of Monoiodoacetic acid.After 8 weeks of oral administration of JWYHD.Western Blot was used to determine the effect of JWYHD on the protein and mRNA expression of ?-catenin,CyclinD1,MMP-3,MMP-13,Caspase-3,Caspase-9 and Col2a1.The protective effects of JWYHD on cartilage were observed by pathological observation with hematoxylin-eosin and Safranning O staining.The serum and synovium pro-inflammatory factors of IL-1 ?,IL-6 and TNF-?of rats were detected by Elisa.Results: The chondroprotective and anti-inflammatory effect of JWYHD on chondrocytes in vitro and on MIA-induced OA rat model in vivo were investigated.In In vitro,JWYHD increased the chondrocyte viability against interleukin(IL)-1?-induced chondrocytes apoptosis and preserved glycosaminoglycans in the extracellular matrix.JWYHD promoted chondrocyte viability against apoptosis,decreased MMP-3,MMP-13,Caspase-3,Caspase-9 via Wnt/?-catenin signaling pathway in both IL-1?-induced and Licl-induced chondrocytes.The qRT-PCR and western blot results showed that mRNA and protein expressions of Wnt signaling pathway related genes ?-catenin and CyclinD1,apoptosis related genes Casapase-3 and Caspase-9,collagen degradation related genes Metalloproteinase(MMP)-3 and MMP-13 were up-regulated,and Col2a1 was down-regulated on IL-1?-induced chondrocytes.Treatment with JWYHD reversed these effects in a dose-dependent manner.Licl was used as Wnt/?-catenin signaling pathway activator in chondrocytes to determine the molecular mechanisms.Activation of Wnt signaling pathway by Licl up-regulated ?-catenin,CyclinD1,Caspase-3,Caspase-9,MMP-3,MMP-13 and IL-1?.These effects were blocked by JWYHD treatment.In vivo,JWYHD decreased the MIA-induced up-regulation of ?-catenin,CyclinD1,Caspase-3,Caspase-9,MMP-3 and MMP-13 protein expressions in cartilage.It was also demonstrated that JWYHD decreased serum and synovium pro-inflammatory cytokines,IL-1?,IL-6 and TNF-? in MIA-induced OA rats and ameliorated the cartilage degradation.Conclusion: JWYHD possess multiple capabilities including preventing chondrocyte apoptosis,preserving integrity of extracellular matrix and anti-inflammatory effect in the treatment of OA both in vitro and in vivo.