药学学报, 2022, 57(4): 1044-1053
引用本文:
李聪翀, 张彦琼, 李玮婕, 毛霞, 刘毓东, 马兆臣, 林雅, 林娜. 从VEGF/VEGFR2/PI3K/AKT信号轴所介导的血管新生调控网络探索白虎加桂枝汤干预类风湿关节炎热证的作用及其机制[J]. 药学学报, 2022, 57(4): 1044-1053.
LI Cong-chong, ZHANG Yan-qiong, LI Wei-jie, MAO Xia, LIU Yu-dong, MA Zhao-chen, LIN Ya, LIN Na. Exploring the effect and mechanism of Baihu-Guizhi Decoction on rheumatoid arthritis with hot syndrome from the angiogenesis regulatory network mediated by VEGF/VEGFR2/PI3K/AKT signaling pathway[J]. Acta Pharmaceutica Sinica, 2022, 57(4): 1044-1053.

从VEGF/VEGFR2/PI3K/AKT信号轴所介导的血管新生调控网络探索白虎加桂枝汤干预类风湿关节炎热证的作用及其机制
李聪翀1,2, 张彦琼2, 李玮婕2, 毛霞2, 刘毓东1,2, 马兆臣2, 林雅1*, 林娜2*
1. 福建中医药大学药学院, 福建 福州 350122;
2. 中国中医科学院中药研究所, 北京 100700
摘要:
出自《金匮要略》的热痹经方白虎加桂枝汤(BHGZD)临床治疗类风湿关节炎(RA)的疗效确切。但针对该方的作用机制研究多围绕调节机体炎症反应和免疫功能等方面,对其抑制滑膜血管新生的药效与机制尚未见报道。本研究采用转录组学数据挖掘、生物网络分析与“动物-细胞”实验验证相整合的研究策略,探讨BHGZD干预RA热证滑膜血管过度新生病理环节的潜能和分子机制。动物福利和实验过程均遵循中国中医科学院实验动物伦理委员会的规定。网络分析结果表明,BHGZD干预RA热证的候选网络靶标显著参与多条血管新生调节相关通路。其中,血管内皮生长因子A-血管内皮生长因子受体2(VEGFA-VEGFR2)信号通路包含多个BHGZD候选网络靶标,如VEGF、磷脂酰肌醇3-激酶(PI3K)、丝-苏氨酸激酶(AKT)等。进一步的实验验证结果表明,BHGZD可有效降低佐剂诱导性关节炎热证大鼠膝关节滑膜中血小板-内皮细胞黏附分子(CD31)的表达,血清中VEGF的水平、内皮性一氧化氮合酶(eNOS)的活性,关节组织中磷酸化血管内皮生长因子受体2(p-VEGFR2)、p-PI3K以及p-AKT蛋白的表达水平,升高血清中内皮抑素(endostatin)的水平,并显著减少HUVEC、MH7A细胞的迁移和侵袭活性,及HUVEC细胞的管腔形成活性。综上所述,BHGZD具有缓解RA热证滑膜血管过度新生的潜能,其作用机制可能与干预VEGF/VEGFR2/PI3K/AKT信号通路相关。
关键词:    白虎加桂枝汤      类风湿关节炎      热证      滑膜血管新生      转录组学      网络药理学     
Exploring the effect and mechanism of Baihu-Guizhi Decoction on rheumatoid arthritis with hot syndrome from the angiogenesis regulatory network mediated by VEGF/VEGFR2/PI3K/AKT signaling pathway
LI Cong-chong1,2, ZHANG Yan-qiong2, LI Wei-jie2, MAO Xia2, LIU Yu-dong1,2, MA Zhao-chen2, LIN Ya1*, LIN Na2*
1. College of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China;
2. Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China
Abstract:
Baihu-Guizhi Decoction (BHGZD), a prescription from ''Synopsis of the Golden Chamber'', has a definite clinical effect in the treatment of rheumatoid arthritis (RA). However, the research on the mechanism of this prescription mainly focuses on the regulation of inflammatory response and immune function, and its efficacy and mechanism of inhibiting synovial angiogenesis have not been reported. In the current study, transcriptomics data mining, biological network analysis and ''in vivo-in vitro'' experimental verification integrated research strategy to explore the potential and molecular mechanism of BHGZD in RA synovial angiogenesis with hot syndrome. Animal welfare and experimental procedures follow the regulations of the Animal Ethics Committee of China Academy of Chinese Medical Sciences. The results of network analysis showed that the candidate network targets of BHGZD intervention in RA with hot syndrome were significantly involved in multiple angiogenesis regulation related pathways. Among them, vascular endothelial growth factor A-vascular endothelial growth factor receptor 2 (VEGFA-VEGFR2) signaling pathway contains multiple BHGZD candidate network targets, such as VEGF, phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), etc. Further experimental results showed that BHGZD could effectively reduce the expression of CD31 in knee synovium, the expression level of VEGF in serum, the activity of endothelial nitric oxide synthase (eNOS), phosphorylated VEGFR2 (p-VEGFR2), p-PI3K and p-AKT in joint tissue of adjuvant-induced arthritis rats with hot syndromes, the migration and invasion activity of HUVEC and MH7A cells, and the lumen formation activity of HUVEC cells and improve the expression level of endostatin in serum. In conclusion, BHGZD has the potential to alleviate excessive synovial angiogenesis in RA with hot syndrome, and its mechanism may be related to the intervention of VEGF/VEGFR2/PI3K/AKT signaling pathway.
Key words:    Baihu-Guizhi Decoction    rheumatoid arthritis    hot syndrome    excessive synovial angiogenesis    transcriptomics    network analysis   
收稿日期: 2021-11-05
DOI: 10.16438/j.0513-4870.2021-1585
基金项目: 国家自然科学基金资助项目(81630107);中国中医科学院科技创新工程资助项目(CI2021A01508).
通讯作者: 林雅,Tel:86-10-64014411-2869,E-mail:80979984@qq.com;林娜,E-mail:linna888@163.com
Email: 80979984@qq.com;linna888@163.com
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