基于VEGF/VEGFR2信号通路探究苗药铁筷子醇提物抗类风湿关节炎大鼠滑膜血管新生作用
DOI:
CSTR:
作者:
作者单位:

贵州中医药大学第二临床医学院

作者简介:

通讯作者:

中图分类号:

基金项目:

国家自然科学基金 (82260874 ),贵州中医药大学2021年国家自然科学基金 后补助资金科研创新探索专项基金 (2018YFC170810524),贵中医大创合字(2023 ) 91号。


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
Author:
Affiliation:

The Second Clinical Medical College of Guizhou University of Traditional Chinese Medicine

Fund Project:

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.

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    【摘要】目的 本项目拟以“苗药铁筷子”为研究对象,研究其对 CIA大鼠抗炎活性及 VEGF/VEGFR2/P38MAPK通路的调控机制。方法 将60只雌性wistar大鼠,随机分为正常组、模型组、阳性药物组、铁筷子低、中、高剂量组,每组10只。将牛II型胶原蛋白溶液注入大鼠尾部,复制大鼠模型,阳性药物组灌胃给予 MTX2.0 mg/(kg·d),每隔一天1次;铁筷子低、中、高三个剂量组以铁筷子乙醇提取物灌胃0.25g/(kg·d)、0.5g/(kg·d)、1g/(kg·d),一天1次;正常组、模型组均采用等量Nacl溶液灌胃;连续灌胃28 d。治疗过程中,观察各组大鼠一般状况,记录体重变化,测定大鼠足底厚度;治疗结束后去大鼠后肢,经microCT检测骨质破坏程度、苏木精-伊红(HE)染色法对关节滑膜的病理学研究,免疫组织化学染色观察滑膜新生血管数量,反转录聚合酶链反应法(qRT-PCR)在大鼠滑膜组织中检测VEGF-A、VEGFR2、TNFα的mRNA水平,蛋白免疫印迹法(Western blot)检测VEGF、VEGFR2、p-p38、AKT的表达,以探索苗药铁筷子治疗类风湿性关节炎的潜在作用机制。结果 与正常组比较,模型组大鼠体质量降低显著(P<0.01),足肿胀度明显增加(P<0.01),病理切片可见踝关节及后肢其他小关节滑膜组织明显增生、大量炎性细胞浸润,骨组织表面可见啃噬样病变,滑膜新生血管数量明显增多,micro-CT显示明显骨质破坏、骨量百分比、骨小梁厚度、骨密度均显著下降,关节滑膜组织中VEGF-A、VEGFR2、TNFα mRNA水平显著升高(P<0.01),VEGF-A、VEGFR2、p-P38、p-AKT蛋白表达量均显著升高(P<0.01)。与模型组比较,铁筷子醇提物呈剂量依赖性改善上述情况,以高剂量组效果最优,大鼠的体质量显著升高(P<0.05),足肿胀度明显下降(P<0.05),滑膜组织病理学损伤及骨组织啃噬样病变情况明显好转,滑膜组织新生血管明显减少,VEGF-A、VEGFR2、TNFα mRNA水平明显降低(P<0.01),VEGF-A、VEGFR2、p-P38、p-AKT蛋白表达量显著降低(P<0.01)。结论 苗药铁筷子可能通过调节VEGF/VEGFR2信号通路减轻 CIA大鼠关节炎性损伤,进而发挥治疗 RA的作用。

    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.

    参考文献
    相似文献
    引证文献
引用本文
分享
相关视频

文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2023-12-04
  • 最后修改日期:2024-04-25
  • 录用日期:2024-10-16
  • 在线发布日期:
  • 出版日期:
文章二维码
防诈骗提示!请勿点击不明链接或添加个人微信。编辑部所有邮箱后缀均为@cnilas.org
关闭