Probing the anti-rheumatoid arthritis synovial neovascularization of alcoholic extracts of the Miao drug helleborus thibetanus franchon in rats based on the VEGF/VEGFR2 signaling pathway
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The Second Clinical Medical College of Guizhou University of Traditional Chinese Medicine

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National Natural Science Foundation of China (82260874 ), Guizhou University of Traditional Chinese Medicine 2021 National Natural Science Foundation of China Post-subsidy Funds for Scientific Research and Innovation Exploration Special Fund (2018YFC170810524), Guizhou University of Traditional Chinese Medicine, Chuanghe Zi (2023 ) No. 91.

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    Abstract:

    【Abstract】Objective This project aims to study the "Miao Medicine helleborus thibetanus franchon " as the research subject, investigating its anti-inflammatory activity in CIA rats and the regulatory mechanism on the VEGF/VEGFR2/P38MAPK pathway. Methods Sixty female Wistar rats were randomly divided into six groups: normal, model, positive control, and low, medium, high dose groups of helleborus thibetanus franchon , with ten rats in each group. Bovine type II collagen solution was injected into the tail of rats to replicate the rat model, and the positive drug group was given MTX2.0 mg/(kg-d) by gavage once every other day; the three dose groups of Iron Chopsticks, low, medium, and high, were gavaged with helleborus thibetanus franchon ethanol extract at 0.25 g/(kg-d), 0.5 g/(kg-d), and 1 g/(kg-d) once a day. The normal and model groups were given an equivalent volume of NaCl solution; continuous administration lasted for 28 days. During treatment, general conditions of the rats were observed, body weight changes recorded, and foot thickness measured. After treatment, the rats' hind limbs were taken for microCT to detect bone destruction, H&E staining for pathological study of the synovial membrane, immunohistochemistry to observe the number of neovascularization in the synovium, qRT-PCR to detect mRNA levels of VEGF-A, VEGFR2, TNFα in rat synovial tissue, and Western blot to detect expressions of VEGF, VEGFR2, p-p38, AKT, exploring the potential mechanism of Miao Medicine helleborus thibetanus franchon in treating rheumatoid arthritis. Results Compared to the normal group, the model group showed significant weight loss (P<0.01), increased foot swelling (P<0.01), visible proliferative synovial tissue with inflammatory cell infiltration, erosive lesions on bone surfaces, increased neovascularization in the synovium, and significant bone destruction in micro-CT, with reduced bone percentage, trabecular thickness, and bone density. Levels of VEGF-A, VEGFR2, TNFα mRNA, and proteins VEGF-A, VEGFR2, p-P38, p-AKT were significantly elevated (P<0.01). Compared to the model group, helleborus thibetanus franchon ethanol extract improved these conditions in a dose-dependent manner, with the high-dose group showing the best effect. There was a significant increase in rats' body weight (P<0.05), reduced foot swelling (P<0.05), ameliorated synovial and erosive bone lesions, reduced neovascularization in the synovium, and significantly lowered levels of VEGF-A, VEGFR2, TNFα mRNA, and proteins VEGF-A, VEGFR2, p-P38, p-AKT (P<0.01). Conclusion: Miao Medicine helleborus thibetanus franchon may alleviate joint inflammatory damage in CIA rats by modulating the VEGF/VEGFR2 signaling pathway, thereby exerting therapeutic effects on RA.

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History
  • Received:December 04,2023
  • Revised:April 25,2024
  • Adopted:October 16,2024
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