ABSTRACT
To study the effect of pulchinenoside (PULC) on the Frizzled (FZD) expression of adjuvant arthritis ( AA) rats. AA rats were prepared through the toe injection with complete Freund's adjuvant to culture fibroblast-like synoviocytes (FLS). The effect of the oral administration with PULC on the FZD8 expression was detected by the real time qPCR. The effect of FZD8 knockout on the expressions of IL-1, IL-6, IL-8 were detected by MTT and ELISA. The role of miR-375 in the abnomal expression of FZD8 was detected by the real time qPCR. The results showed signfiicant decrease in the FZD8 expression among AA rats, FLS proliferation ater FZD8 knockout and IL-1, IL-6, IL-8 expressions and notable increase in miR-375 expression after the oral administration with PULC. The up-regulated miR-375 expression can inhibit the FZD8 expression. PULC may inhibit the FZD8 expression by up-regulating the miR-375 expression.
Subject(s)
Arthritis, Experimental/drug therapy , Drugs, Chinese Herbal/administration & dosage , Receptors, Cell Surface/genetics , Saponins/administration & dosage , Animals , Arthritis, Experimental/genetics , Arthritis, Experimental/metabolism , Disease Models, Animal , Humans , Male , Rats , Rats, Sprague-Dawley , Receptors, Cell Surface/metabolismABSTRACT
The role of pulchinenoside (PULC) in the regulation of MeCP2 expression was investigated in RA model rats. Adjuvant arthritis rats were used as RA model rats, and fibroblast-like synoviocytes (FLS) from the RA model rats were cultured. The effect of 100 mg x kg(-1) PULC gavage treatment on the MeCP2 expression and the effect of MeCP2 siRNA on the expression of SFRP2 and ß-catenin were detected by real time qPCR and Western blotting. The role of PULC in the FLS proliferation was detected by MTT. The results showed that the MeCP2 expression was down-regulated, the SFRP2 expression was up-regulated and the FLS proliferation was inhibited in FLS after therapy. MeCP2 siRNA significantly inhibited the MeCP2 expression, up-regulated the SFRP2 expression and inhibited the ß-catenin expression in FLS from RA model rats. PULC may increase the SFRP2 expression, inhibit the Wnt signaling and inhibit the FLS proliferation in FLS from the RA model rats by inhibiting the MeCP2 expression.
Subject(s)
Arthritis, Rheumatoid/drug therapy , Arthritis, Rheumatoid/genetics , Drugs, Chinese Herbal/administration & dosage , Fibroblasts/metabolism , Methyl-CpG-Binding Protein 2/genetics , Animals , Arthritis, Rheumatoid/metabolism , Disease Models, Animal , Fibroblasts/drug effects , Gene Expression Regulation/drug effects , Humans , Male , Methyl-CpG-Binding Protein 2/metabolism , Rats , Rats, Sprague-Dawley , Synovial Membrane/cytology , Synovial Membrane/drug effects , Synovial Membrane/metabolism , Wnt Signaling Pathway/drug effects , beta Catenin/genetics , beta Catenin/metabolismABSTRACT
OBJECTIVE: To study the effect of pulchinenoside (PULC) in modulating SFRP2 expression in fibroblast-like synoviocytes (FLS) of rheumatoid arthritis (RA) model rats. METHOD: The effect of PULC in treating RA rats was evaluated by rat arthritis score and paw swelling score. The inhibitory effect of PULC on FLS proliferation was detected by MTT reagent. The effects of PULC gavage treatment in modulating gene expression of FLS SFRP2, critical gene beta-catenin of Wnt pathway and downstream effector genes C-myc of of Wnt pathway were detected by RT-PCR and Western blotting. RESULT: PULC had a significant effect in treating RA rats and that SFRP2 expression was down-regulated in FLS. After PULC gavage treatment, FLS SFRP2 expression was obviously up-regulated, whereas beta-catenin and C-myc gene expressions were significantly down-regulated. CONCLUSION: PULC can inhibit abnormal proliferation of synovial membrane by modulating Wnt pathway of RA rats.
