Combination of Components from Tripterygii Radix et Rhizoma-Chuanxiong Rhizoma Affects RA-FLSs by Regulating NF-κB,Nrf2/HO-1 Signaling Pathways and Bcl-2/Caspase-3 Expression
Objective:To investigate the effects of the combination of components from Tripterygii Radix et Rhizoma and Chuanxiong Rhizoma on rheumatoid arthritis fibroblast-like synoviocytes(RA-FLSs)and the underlying mechanism.Methods:RA-FLSs were grouped as follows:blank control,positive control(methotrexate),Tripterygii Radix et Rhizoma components,Chuanxiong Rhizoma components,and components from Tripterygii Radix et Rhizoma+Chuanxiong Rhizoma.The cell-counting kit-8(CCK-8)assay was employed to the cell proliferation,invasion,and apoptosis.The levels of tumor necrosis factor(TNF)-α,interleukin(IL)-6,reactive oxygen species(ROS),and malondiadehyde(MDA)in cells were measured.Western blot was employed to determine the protein levels of nuclear factor erythroid 2-related factor 2(Nrf2),heme oxygenase-1(HO-1),nuclear factor-kappa B(NF-κB)p65,phosphorylated inhibitory subunit of NF-κBα(p-IKBα),cysteinyl aspartate-specific protease-3(Caspase-3),and B-cell lymphoma 2(Bcl-2).Real-time PCR was employed to determine the mRNA levels of Nrf2,HO-1,and NF-κB p65.Results:The cells in the groups of positive control,Tripterygii Radix et Rhizoma components,Chuanxiong Rhizoma components,and components from Tripterygii Radix et Rhizoma+Chuanxiong Rhizoma were treated with 2.50 mg·L-1 methotrexate,0.20 mg·L-1 triptolide+0.20 mg·L-1 celastrol,5.00 mg·L1 ferulic acid+20.00 mg·L-1 ligustrazine,0.20 mg·L-1 triptolide+0.20 mg·L-1 celastrol+5.00 mg·L-1 ferulic acid+20.00 mg·L-1 ligustrazine,respectively.Compared with the blank control group,drug administration reduced the proliferation and invasion and increased the apoptosis of cells(P＜0.01),lowered the levels of TNF-α,IL-6,ROS,and MDA(P＜0.01),up-regulated the mRNA and protein levels of Caspase-3,Nrf2,and HO-1(P＜0.01),and down-regulated the mRNA and protein levels of Bcl-2,NF-κB p65,and p-IKBα(P＜0.01).Compared with the Tripterygii Radix et Rhizoma components group,the combination of components from Tripterygii Radix et Rhizoma+Chuanxiong Rhizoma inhibited the proliferation and invasion(P＜0.05)and promoted the apoptosis of RA-FLSs,up-regulated the mRNA levels of Nrf2 and HO-1 and protein levels of Nrf2 and Caspase-3(P＜0.05),and down-regulated the protein levels of NF-κB p65 and p-IκBα(P＜0.05).Conclusion:The combination of components from Chuanxiong Rhizoma and Tripterygii Radix et Rhizoma can inhibit the proliferation and invasion and promote the apoptosis of RA-FLSs and alleviate oxidative stress and inflammation by inhibiting the NF-κB signaling pathway,activating the Nrf2/HO-1 pathway,and regulating the expression of Bcl-2/Caspase-3.
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