Effects of Catalpol on the Proliferation of Rheumatoid Arthritis Fibroblast Synovial Cells and YAP/TAZ Signaling Pathway
This article explores the effects of catalpol on proliferation of FLS(fibroblast-like synoviocytes)and YAP(yes-associated protein)/TAZ(transcriptional co-activator with PDZ-binding motif)signaling pathway in RA(rheumatoid arthritis).Human arthritis fibroblast like synovial cells HFLS-RA were cultured in vitro and induced with 20 ng/mL TNF-α(tumor necrosis factor-α)to establish a RA cell model,SD rats were selected to con-struct an RA animal model using type Ⅱ collagen,and then randomly grouped into model group,catalpol group,empty group,and catalpol+YAP overexpression group,10 per group,another 10 SD rats and normal cultured HFLS-RA cells were selected as the control group,after grouping and treatment with catalpol,empty plasmid,and YAP overexpression plasmid,CCK-8 method and TUNEL staining,flow cytometry were applied to detect the pro-liferation and apoptosis of HFLS-RA cells in each group,respectively;the foot volume of rats in each group was measured by the toe measuring instrument and the arthritis score was performed according to the 5-level scoring method;TUNEL staining was applied to detect apoptosis in the synovial tissue of rats in each group;ELISA(en-zyme-linked immunosorbent assay)was applied to detect the levels of pro-inflammatory factors IL-1β(interleukin-1β),MCP-1(monocyte chemotactic protein-1),and IL-6 in serum of HFLS-RA cells and RA rats in each group;immunoblotting experiments were applied to detect the expression of proliferation and YAP/TAZ signaling related proteins in HFLS-RA cells and synovial tissue of RA rats in each group.The results showed that,compared with the control group,the survival rate of HFLS-RA cells,rat foot volume and arthritis score,the levels of IL-6,MCP-1,IL-1β in HFLS-RA cells and in serum of RA rats,and the expression of Cyclin D1,YAP and TAZ proteins in HFLS-RA cells and in synovial tissue of RA rats in the model group increased(P＜0.05),while the apoptosis rate of HFLS-RA cells and the apoptosis ratio of synovial tissue cells in rat decreased(P＜0.05).Compared with the model group,the survival rate of HFLS-RA cells,rat foot volume and arthritis score,the levels of IL-6,MCP-1,IL-1β in HFLS-RA cells and in serum of RA rats,and the expression of Cyclin D1,YAP and TAZ proteins in HFLS-RA cells and in synovial tissue of RA rats in the catalpol group decreased(P＜0.05),the apoptosis rate of HFLS-RA cells and the apoptosis ratio of synovial tissue cells in rat increased(P＜0.05);there was no significant change in all indicators in the empty load group(P＞0.05).Compared with the catalpol group,the survival rate of HFLS-RA cells,rat foot volume and arthritis score,the levels of IL-6,MCP-1,IL-1β in HFLS-RA cells and in serum of RA rats,and the expression of Cyclin D1,YAP and TAZ proteins in HFLS-RA cells and in synovial tissue of RA rats in the catalpol+YAP overexpression group increased(P＜0.05),while the apoptosis rate of HFLS-RA cells and the apop-tosis ratio of synovial tissue cells in rat decreased(P＜0.05).In short,catalpol can reduce inflammation levels in RA rats and cell models by weakening YAP/TAZ signaling activity,thereby inhibiting FLS proliferation and alleviating arthritis symptoms in RA rats.
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