Abstract
Objective
To investigate the effect of isorhamnetin on the pathology of rheumatoid arthritis (RA).
Methods
Tumor necrosis factor (TNF)- α -induced fibroblast-like synoviocytes (FLS) was exposed to additional isorhamnetin (10, 20 and 40 µ mol/L). Overexpression vectors for matrix metalloproteinase-2 (MMP2) or MMP9 or SRC were transfected to explore their roles in isorhamnetin-mediated RA-FLS function. RA-FLS viability, migration, and invasion were evaluated. Moreover, a collagen-induced arthritis (CIA) rat model was established. Rats were randomly divided to sham, CIA, low-, medium-, and high-dosage groups using a random number table (n=5 in each group) and administed with normal saline or additional isorhamnetin [2, 10, and 20 mg/(kg·day)] for 4 weeks, respectively. Arthritis index was calculated and synovial tissue inflammation was determined in CIA rats. The levels of MMP2, MMP9, TNF-α, interleukin-6 (IL-6), and IL-1 β, as well as the phosphorylation levels of SRC, extracellular regulated kinase (ERK), and cyclic adenosine monophosphate response element-binding (CREB), were detected in RA-FLS and synovial tissue. Molecular docking was also used to analyze the binding of isorhamnetin to SRC.
Results
In in vitro studies, isorhamnetin inhibited RA-FLS viability, migration and invasion (P<0.05). Isorhamnetin downregulated the levels of MMP2, MMP9, TNF-α, IL-6, and IL-1 β in RA-FLS (P<0.05). The overexpression of either MMP2 or MMP9 reversed isorhamnetin-inhibited RA-FLS migration and invasion, as well as the levels of TNF-α, IL-6, and IL-1 β (P<0.05). Furthermore, isorhamnetin bound to SRC and reduced the phosphorylation of SRC, ERK, and CREB (P<0.05). SRC overexpression reversed the inhibitory effect of isorhamnetin on RA-FLS viability, migration and invasion, as well as the negative regulation of MMP2 and MMP9 (P<0.05). In in vivo studies, isorhamnetin decreased arthritis index scores (P<0.05) and alleviated synovial inflammation. Isorhamnetin reduced the levels of MMP2, MMP9, TNF-α, IL-6, and IL-1 β, as well as the phosphorylation of SRC, ERK, and CREB in synovial tissue (P<0.05). Notably, the inhibitory effect of isorhamnetin was more pronounced at higher concentrations (P<0.05).
Conclusion
Isorhamnetin exhibited anti-RA effects through modulating SRC/ERK/CREB and MMP2/MMP9 signaling pathways, suggesting that isorhamnetin may be a potential therapeutic agent for RA.
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Availability of Data and Material
The datasets used and analyzed during the current study available from the corresponding author on reasonable request.
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Li XR contributions to conceptualization, data curation, investigation, methodology, funding acquisition, and writing of the original draft. Li SF and Mei WY contributed to conceptualization, formal analysis, validation, investigation, software, and validation. Liu XD and Zhou RB contributed to conceptualization, project administration, supervision, and review. All authors read and approved the final manuscript.
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Supported by the Natural Science Foundation of Hunan Province (Nos. 2022JJ80086 and 2023JJ60342), the Project of Hunan Provincial Health and Health Commission (No. D202302078705), the Project of Hunan Provincial Student Innovation and Entrepreneurship Training Program (No. 2022–5313), the Hunan Provincial Administration of Traditional Chinese Medicine Scientific Research Program (No. 2021161), the Hunan University of Traditional Chinese Medicine Primary Discipline Open Fund Project in Chinese Medicine (No. 2020ZYX01), the Key Discipline Project on Chinese Pharmacology of Hunan University of Chinese Medicine (No. 202302), and the Scientific Research Project of Hunan Provincial Administration of Traditional Chinese Medicine (No. B2023150)
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Liu, Xr., Li, Sf., Mei, Wy. et al. Isorhamnetin Downregulates MMP2 and MMP9 to Inhibit Development of Rheumatoid Arthritis through SRC/ERK/CREB Pathway. Chin. J. Integr. Med. 30, 299–310 (2024). https://doi.org/10.1007/s11655-023-3753-6
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DOI: https://doi.org/10.1007/s11655-023-3753-6