Liu Yuwen, Liu Jiping, Hu Naping, Li Zhengrong, Liu Anqi, Luo Ruyue, Du Siyu, Guo Dongyan, Li Jiankang, Duan Jialin
Xi'an Key Laboratory of Stem Cell and Regenerative Medicine, Institute of Medical Research, Northwestern Polytechnical University, 127 West Youyi Road, Xi'an, Shaanxi, 710072, China.
The Key Laboratory of Basic and New Drug Research of Traditional Chinese Medicine, Shaanxi University of Chinese Medicine, Xi'an, 712046, Shaanxi, China.
J Ethnopharmacol. 2025 Jan 31;340:119300. doi: 10.1016/j.jep.2024.119300. Epub 2024 Dec 28.
The Daqinjiao decoction (DQJT), a classical prescription, has been utilized for millennia in stroke management, yet its underlying mechanisms remained obscure.
The aim of this study was to elucidate the mechanisms through which DQJT mitigates cerebral ischemia/reperfusion injury (CI/RI).
The quantification of DQJT's primary components were performed by HPLC. Pharmacological assessments were then conducted to ascertain DQJT's efficacy in a Middle Cerebral Artery Occlusion/Reperfusion (MCAO/R) model. Following this, untargeted metabolomics, lipidomics and network pharmacology analyses were undertaken to unveil potential mechanisms, which were subsequently validated. UPLC-Q-TOF/MS was utilized to detect DQJT-derived chemicals in brain tissue, and molecular docking techniques were employed to investigate the bioactive compounds.
DQJT treatment reduced brain damage induced by MCAO/R, as evidenced by decreased infarct sizes, enhanced behavioral function scores, and diminished neuronal damages. Untargeted metabolomics and lipidomics revealed that DQJT improved metabolism of unsaturated fatty acids. According to network pharmacology, lipid metabolism, cAMP signaling pathway and toll-like receptor signaling pathway pathways were notably affected, with HSP90AA1, TLR4, and PKA identified as potential targets of DQJT. Immunofluorescence and Western blot analyses further demonstrated that DQJT counteracted necroptosis and ferroptosis by inhibiting the HSP90AA1 and TLR4 pathways and enhancing the PKA pathway. Molecular docking results supported that the possible pharmacodynamic substances of DQJT in protecting against CI/RI.
This research established that DQJT attenuates brain injury induced by MCAO/R through the modulation of necroptosis and ferroptosis via pathways including HSP90AA1, TLR4, and PKA. It shed light on the potential mechanisms and effective constituents of DQJT in stroke treatment, paving the way for further exploration of this ancient formula.
大秦艽汤（DQJT）作为一个经典方剂，在中风治疗中已应用了数千年，但其潜在机制仍不清楚。
本研究旨在阐明大秦艽汤减轻脑缺血/再灌注损伤（CI/RI）的机制。
采用高效液相色谱法对大秦艽汤的主要成分进行定量分析。然后进行药理学评估，以确定大秦艽汤在大脑中动脉闭塞/再灌注（MCAO/R）模型中的疗效。在此之后，进行非靶向代谢组学、脂质组学和网络药理学分析，以揭示潜在机制，随后进行验证。采用超高效液相色谱-四极杆飞行时间质谱联用仪（UPLC-Q-TOF/MS）检测脑组织中源自大秦艽汤的化学物质，并采用分子对接技术研究生物活性化合物。
大秦艽汤治疗可减轻MCAO/R诱导的脑损伤，表现为梗死面积减小、行为功能评分提高和神经元损伤减轻。非靶向代谢组学和脂质组学研究表明，大秦艽汤可改善不饱和脂肪酸代谢。根据网络药理学分析，脂质代谢、环磷酸腺苷（cAMP）信号通路和Toll样受体信号通路受到显著影响，热休克蛋白90α家族成员1（HSP90AA1）、Toll样受体4（TLR4）和蛋白激酶A（PKA）被确定为大秦艽汤的潜在靶点。免疫荧光和蛋白质印迹分析进一步表明，大秦艽汤通过抑制HSP90AA1和TLR4信号通路以及增强PKA信号通路来对抗坏死性凋亡和铁死亡。分子对接结果支持大秦艽汤中可能的药效物质对CI/RI具有保护作用。
本研究证实，大秦艽汤通过调节包括HSP90AA1、TLR4和PKA在内的信号通路的坏死性凋亡和铁死亡，减轻MCAO/R诱导的脑损伤。本研究揭示了大秦艽汤在中风治疗中的潜在机制和有效成分，为进一步探索这一古老方剂铺平了道路。
