Integrating serum pharmacochemistry, network pharmacology, and metabolomics to explore the protective mechanism of Hua-Feng-Dan in ischemic stroke

IF 8.3 1区医学 Q1 CHEMISTRY, MEDICINAL

Phytomedicine Pub Date : 2025-09-10 DOI:10.1016/j.phymed.2025.157251

Xiaofeng Yuan , Xiaoxia He , Qilin Shu , Yayang Gao , Youli Chen , Jian Xu , Yongping Zhang , Guoqiong Cao

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Abstract

Background

The traditional Chinese medicine Hua-Feng-Dan (HFD) has shown efficacy against ischemic stroke, but how it works remains unclear.

Purpose

To elucidate the mechanisms of action of HFD against ischemic stroke.

Methods

The effects of HFD on ischemic stroke injury were explored in rats subjected to middle cerebral artery occlusion/reperfusion (MCAO/R). The components of HFD and in serum was determined after oral administration using ultra-high performance liquid chromatography followed by mass spectrometry. Potential therapeutic targets of the HFD components in serum were identified through the combination of network pharmacology and metabolomic analysis of serum, then validated through molecular docking and surface plasmon resonance(SPR), as well as immunohistochemistry and western blotting of brain tissue.

Results

HFD alleviated nerve function injury, damage to cortical and hippocampal neurons, and oxidative stress induced by MCAO/R, while also shrinking cerebral infarctions and dampening inflammatory responses and neuronal apoptosis. The main ingredients of HFD are flavonoids and alkaloids. Network pharmacology based on chemical components in serum, followed by molecular docking and SPR, identified several potential therapeutic targets, including AKT1 and PIK3CA. Western blotting of brain tissue confirmed that HFD activated PI3K/AKT/mTOR signaling. Metabolomic analysis of serum revealed that HFD helped to renormalize stroke-perturbed metabolic pathways involving glycerophospholipids, arachidonic acid, primary bile acids and tryptophan. It also revealed that HFD downregulated ACHE, PTGS2, CYP19A1 and ALOX5, which immunohistochemistry of brain tissue confirmed.

Conclusion

HFD exerts therapeutic effects in a rat model of ischemic stroke through multiple pathways and targets, including PI3K/AKT/mTOR signaling; metabolic pathways involving arachidonic and amino acids; and the enzymes encoded by ACHE, MAOA, PTGS2, CYP19A1, and ALOX5. These insights may guide further studies into the mechanisms of HFD, which may help optimize its formulation or lead to next-generation treatments based on individual components.

Abstract Image

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结合血清药物化学、网络药理学、代谢组学等方法探讨花风丹对缺血性脑卒中的保护作用机制。

背景：中药化风丹（HFD）对缺血性中风有一定的疗效，但其作用机理尚不清楚。目的：探讨HFD对缺血性脑卒中的作用机制。方法：探讨HFD对大脑中动脉闭塞/再灌注（MCAO/R）大鼠缺血性脑卒中损伤的影响。采用超高效液相色谱-质谱联用法测定口服给药后血清中HFD和HFD的成分。通过网络药理学和血清代谢组学分析相结合，确定血清中HFD成分的潜在治疗靶点，然后通过分子对接和表面等离子体共振（SPR），以及脑组织免疫组织化学和western blotting进行验证。结果：HFD可减轻MCAO/R诱导的神经功能损伤、皮质和海马神经元损伤以及氧化应激，同时可缩小脑梗死灶，抑制炎症反应和神经元凋亡。HFD的主要成分是类黄酮和生物碱。基于血清化学成分的网络药理学，然后是分子对接和SPR，确定了几个潜在的治疗靶点，包括AKT1和PIK3CA。脑组织Western blotting证实HFD激活了PI3K/AKT/mTOR信号通路。血清代谢组学分析显示，HFD有助于重新规范中风紊乱的代谢途径，包括甘油磷脂、花生四烯酸、原胆汁酸和色氨酸。脑组织免疫组化证实，HFD下调了ACHE、PTGS2、CYP19A1和ALOX5。结论：HFD在缺血性脑卒中大鼠模型中通过多种途径和靶点发挥治疗作用，包括PI3K/AKT/mTOR信号通路；涉及花生四烯和氨基酸的代谢途径；以及由ACHE、MAOA、PTGS2、CYP19A1和ALOX5编码的酶。这些见解可能指导对HFD机制的进一步研究，这可能有助于优化其配方或基于单个成分的下一代治疗方法。

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来源期刊

Phytomedicine 医学-药学

CiteScore

10.30

自引率

5.10%

发文量

670

审稿时长

91 days

期刊介绍： Phytomedicine is a therapy-oriented journal that publishes innovative studies on the efficacy, safety, quality, and mechanisms of action of specified plant extracts, phytopharmaceuticals, and their isolated constituents. This includes clinical, pharmacological, pharmacokinetic, and toxicological studies of herbal medicinal products, preparations, and purified compounds with defined and consistent quality, ensuring reproducible pharmacological activity. Founded in 1994, Phytomedicine aims to focus and stimulate research in this field and establish internationally accepted scientific standards for pharmacological studies, proof of clinical efficacy, and safety of phytomedicines.