WTD Attenuating Rheumatoid Arthritis via Suppressing Angiogenesis and Modulating the PI3K/AKT/mTOR/HIF-1α Pathway

Abstract

Background: Wutou Decoction (WTD), as a classic prescription, has been generally used to treat rheumatoid arthritis (RA) for two thousand years in China. However, the potential protective effects of WTD on rheumatoid arthritis and its possible mechanism have rarely been reported.Purpose: The aim of this study was to explore the possible mechanism of WTD against RA and a promising alternative candidate for RA therapy.Methods: A model of collagen-induced arthritis (CIA) was constructed in rats to assess the therapeutic effects of WTD. Histopathological staining, immunofluorescence, and western blotting of synovial sections were conducted to detect the antiangiogenic effects of WTD. Then, cell viability assays, flow cytometry, scratch healing assays, and invasion assays were conducted to explore the effects of WTD on MH7A human fibroblast-like synoviocyte (FLS) cell proliferation, apoptosis, migration, and invasion in vitro. The ability of WTD to induce blood vessel formation after MH7A cell and human umbilical vein endothelial cell line (HUVEC) coculture with WTD intervention was detected by a tube formation assay. The mechanisms of WTD were screened by network pharmacology and confirmed by in vivo and in vitro experiments.Results: WTD ameliorated the symptoms and synovial pannus hyperplasia of CIA rats. Treatment with WTD inhibited MH7A cell proliferation, migration, and invasion and promoted MH7A apoptosis. WTD could inhibit MH7A cell expression of proangiogenic factors, including VEGF and ANGI, to induce HUVEC tube formation. Furthermore, the PI3K-AKT-mTOR-HIF-1α pathway was enriched as a potential target of WTD for the treatment of RA through network pharmacology enrichment analysis. Finally, it was confirmed in vitro and in vivo that WTD inhibits angiogenesis in RA by interrupting the PI3K-AKT-mTOR-HIF-1α pathway.Conclusion: WTD can inhibit synovial hyperplasia and angiogenesis, presumably by inhibiting the migration and invasion of MH7A cells and blocking the production of proangiogenic effectors in MH7A cells. The possible underlying mechanism by which WTD ameliorates angiogenesis in RA is the PI3K-AKT-mTOR-HIF-1α pathway.

Gancao Nourishing-Yin decoction combined with methotrexate in treatment of aging CIA mice: a study based on DIA proteomic analysis

“…The inhibition of various inflammation factors is also the most significant, such as Stat1, Hspb6 and Tgfb. Pi3k-Akt signaling pathway has been profoudly investigated within RA by promoting angiogenesis and activation of fibroblast-like synoviocytes, and testified as reliable therapeutic target [ 19 , 31 , 32 ]. Besides, it participates in aging and ARDs by crosstalk with oxidative stress, DNA damage, apoptosis and inflammation [ 33 , 34 ].…”

Signaling pathways in rheumatoid arthritis: implications for targeted therapy

“…225 Abnormal PI3K/ AKT pathway activation will also stimulate the expression of VEGF and HIF-1α to promote angiogenesis, which not only isolates bones from getting nutrients through the synovium but also gets involved in the release of diverse inflammatory mediators, aggravating the condition of RA. [226][227][228] There is evidence indicating that the PI3K/AKT/mTOR pathway participates in the process of RA, the mammalian target of rapamycin (mTOR) inhibits autophagy in FLS, promotes continuous abnormal proliferation of synovial cells, and is also critical for the survival and differentiation of osteoclasts, aggravating RA and mTOR might be a target for RA or other autoimmune diseases. 227,[229][230][231] SYK signaling pathway in RA SYK (spleen tyrosine kinase) is a central molecule of B-cell receptor signaling.…”

Biology of interleukin‑37 and its role in autoimmune diseases (Review)

“…Downregulation of hypoxia-inducible factor-1α (HIF-1α) significantly reduces angiogenesis in VEGF-induced rheumatoid arthritis fibroblast-like synoviocytes (RAFLS) in macrophages of the synovial lining ( 60 ). Multiple studies have shown that blocking angiogenic pathways may reduce inflammatory cell infiltration and damage to joints ( 61 , 62 ). In animal models of RA, prophylactic treatment with anti-VEGF antibodies delays the onset, joint swelling, and vascularization in collagen-induced arthritis (CIA) ( 63 ).…”