Li Jia, Zhang Xiaofei, Guo Dongyan, Shi Yajun, Zhang Shihao, Yang Ruiying, Cheng Jiangxue
Department of Pharmaceutics, The Key Laboratory of Basic and New Drug Research of Traditional Chinese Medicine, Shaanxi University of Chinese Medicine, Xianyang, China.
Department of Pharmaceutics, The Key Laboratory of Modern Preparation of Traditional Chinese Medicine, Ministry of Education, Jiangxi University of Chinese Medicine, Nanchang, China.
Front Pharmacol. 2023 Mar 13;14:1113810. doi: 10.3389/fphar.2023.1113810. eCollection 2023.
To investigate the effects and mechanisms of Paeoniae radix rubra-Angelicae sinensis radix (P-A) drug pair in the treatment of rheumatoid arthritis (RA). Mass spectrometry was employed to accurately characterize the main components of the P-A drug pair. Network pharmacology was used to analyze the main components and pathways of the P-A drug pair in the treatment of RA, and Discovery Studio software was used to molecularly dock the key proteins on the pathway with their corresponding compounds. The levels of serum TNF-a, IL-1β, and IL-6 were measured by enzyme linked immunosorbent assay (ELISA). The histopathology of the ankle joint was observed by hematoxylin-eosin (HE) staining, and the positive expression of p-PI3K, p-IKK, p-NF-κB, and p-AKT in the synovial tissue of the ankle joint was detected by immunohistochemical analysis. Finally, the expression of PI3K, IKK, and AKT and their phosphorylation levels were determined by western blot in each group of rats. Network pharmacology combined with molecular docking analysis revealed that the pharmacodynamic mechanism of the P-A drug pair for the treatment of RA may be related to the contents of caffeic acid, quercetin, paeoniflorin, and baicalein in the regulation of the expression of the PI3K/AKT/NF-κB signaling pathway and the targets of PIK3CA, PIK3R1, AKT1, HSP90AA1 and IKBKB in the pathway. Compared with the model group, the P-A drug pair significantly improved the pathological changes of the synovial tissue and reduced feet swelling in RA model rats. Moreover, it regulated the levels of TNF-α, IL-1β, and IL-6 in serum ( < 0.05). The results of the immunohistochemical analysis and western blot showed that the expression of PI3K, IKK, NF-κB, and AKT decreased after phosphorylation in the synovial tissue ( < 0.05). The P-A drug pair exhibited an inhibitory effect on the hyperactivation of the PI3K/AKT/NF-κB signaling pathway in the synovial membrane of RA rats. The mechanism may be related to the downregulation of the phosphorylation levels PI3K, IKK, NF-κB, and AKT, which in turn decreased inflammatory cell infiltration and synovial membrane proliferation.
探讨赤芍 - 当归药对治疗类风湿关节炎（RA）的作用及机制。采用质谱法准确表征赤芍 - 当归药对的主要成分。运用网络药理学分析赤芍 - 当归药对治疗RA的主要成分及通路，并使用Discovery Studio软件对通路中的关键蛋白与其相应化合物进行分子对接。采用酶联免疫吸附测定（ELISA）法检测血清TNF -α、IL -1β和IL -6水平。通过苏木精 - 伊红（HE）染色观察踝关节组织病理学变化，采用免疫组织化学分析法检测踝关节滑膜组织中p - PI3K、p - IKK、p - NF -κB和p - AKT的阳性表达。最后，通过蛋白质免疫印迹法检测每组大鼠中PI3K、IKK和AKT的表达及其磷酸化水平。网络药理学结合分子对接分析表明，赤芍 - 当归药对治疗RA的药效机制可能与咖啡酸、槲皮素、芍药苷和黄芩苷调节PI3K/AKT/NF -κB信号通路表达的含量以及该通路中PIK3CA、PIK3R1、AKT1、HSP90AA1和IKBKB的靶点有关。与模型组相比，赤芍 - 当归药对显著改善了RA模型大鼠滑膜组织的病理变化，减轻了足肿胀。此外还调节了血清中TNF -α、IL -1β和IL -6的水平（P<0.05）。免疫组织化学分析和蛋白质免疫印迹结果显示，滑膜组织中PI3K、IKK、NF -κB和AKT磷酸化后表达降低（P<0.05）。赤芍 - 当归药对在RA大鼠滑膜中对PI3K/AKT/NF -κB信号通路的过度激活具有抑制作用。其机制可能与PI3K、IKK、NF -κB和AKT磷酸化水平下调有关，进而减少炎症细胞浸润和滑膜增殖。
