Effect of Bushen Huoxue formula regulating NF-κB signaling pathway on inflammation and apoptosis of cartilage tissue in mice with knee osteoarthritis
Objective To determine the mechanism of Bushen Huoxue formula(BSHXF)for treatment of knee osteoarthritis(KOA)and to observe the effect of BSHXF on the apoptosis and inflammation of KOA in mice.Methods Totally 24 male BALB/c mice were stratified by body weight and randomly divided into a sham group,a KOA group and a KOA+BSHXF group(8 mice in each group).After the intervention of model building and intragastric administration,the serum was collected and the levels of IL-6,iNOS and COX2 were detected by ELISA.The cartilage tissues of the mice were collected,and the histopathology of the knee cartilage was detected by HE staining and saffranine O-solid green staining.TUNEL staining was used to detect the apoptosis of the cartilage tissues.The protein expression of Cleaved-Caspase-9,Bcl-2,Bax and p-IκBα,IκBα,p-p65 and p65,namely the key proteins in NF-κB signaling pathway,were detected by Western blot,and the mRNA expressions of Collagen Ⅱ,Aggrecan and ADAMTS-5 were detected by qRT-PCR.Results Compared with the sham group,the pathological changes in the knee cartilage cells in the KOA group were obvious.The serum levels of inflammatory cytokines IL-6,iNOS and COX2 were increased.The apoptosis rate in the cartilage tissue,Cleaved-Caspase-9 and Bax protein expression levels were also increased.Bcl-2,Collagen Ⅱ and Aggrecan mRNA levels were decreased,while ADAMTS-5 mRNA,the ratios of p-IκBα/IκBα and p-p65/p65 were increased.Compared with the KOA group,injurying the knee joint cartilage and pathological changes were improved the levels of inflammatory cytokines IL-6,iNOS and COX2,apoptosis rate in the cartilage tissue,the protein expression levels of Cleaved-Caspase-9 and Bax were all decreased,but Bcl-2,the mRNA levels of Collagen Ⅱ and Aggrecan were increased,while ADAMTS-5 mRNA and the ratios of p-IκBα/IκBα and p-p65/p65 were decreased in the KOA+BSHXF group.Conclusion BSHXF may inhibit NF-κB signaling pathway,reduce the release of inflammatory factors,inhibit chondrocyte apoptosis and extracellular matrix degradation,to alleviate the progression of KOA.
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