Buyang Huanwu Decoction Alleviates Ischemic Stroke Injury by Inhibiting Ferroptosis via the Nrf2/GPX4 Pathway.

IF 4.7 2区医学 Q1 CHEMISTRY, MEDICINAL

Drug Design, Development and Therapy Pub Date : 2025-03-27 eCollection Date: 2025-01-01 DOI:10.2147/DDDT.S503424

Hao Huang, Sijie Liu, Jing Wu, Jiayi Zhu, Jiaxiang Xu, Shuhong Yu, Lingna Bei, Biao Zhang, Yi Luo

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引用次数: 0

Abstract

Purpose: Acute ischemic stroke poses major challenges due to high disability and mortality rates. Ferroptosis, a form of regulated cell death triggered by iron-induced oxidative stress, plays a key role in stroke injury. Despite its long history in stroke treatment, the mechanism of Buyang Huanwu Decoction (BHD) in ferroptosis remains unclear.

Methods: Network pharmacology predicted BHD's active components and pathways, while Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS/MS) confirmed its main ingredients. Middle Cerebral Artery Occlusion (MCAO) was induced in C57 mice, with neurological deficits and infarct size assessed by Longa scoring and TTC staining. Histopathological and ultrastructural changes were assessed by staining and electron microscopy, and biochemical markers (MDA, GSH, SOD, Fe²⁺) measured by kits. Western blotting and qPCR analyzed ferroptosis-related proteins, Nrf2 localization, and gene expression. In vitro, HT22 cells viability and ROS levels were assessed under Oxygen-Glucose Deprivation/Reoxygenation (OGD/R) conditions. Protein expression, Nrf2 interactions, and nuclear translocation were also investigated.

Results: Network pharmacology showed BHD targets key pathways in cerebral infarction, including ferroptosis and antioxidant pathways. BHD improved neurological function and reduced the infarct size in MCAO mice by 10% - 50%, and also significantly decreased the levels of oxidative stress markers (MDA, Fe²⁺) while increasing the activities of antioxidants (GSH, SOD). Histopathological and ultrastructural analyses demonstrated reduced neuronal damage and improved mitochondrial structure. Western blot and qPCR indicated upregulation of GPX4 and Nrf2, downregulation of Keap1, and Nrf2 nuclear translocation. In vitro, BHD enhanced HT22 cell viability and reduced ROS under stress. Protein analysis confirmed increased Nrf2, GPX4, and HO-1, with decreased Keap1 and enhanced Nrf2 nuclear translocation. Nrf2 inhibitor experiments confirmed BHD's effects are Nrf2-mediated.

Conclusion: In pre-clinical studies, BHD exerts neuroprotective effects in ischemic stroke by inhibiting ferroptosis through the Nrf2/GPX4 pathway.

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补阳还五汤通过Nrf2/GPX4通路抑制铁下沉减轻缺血性脑卒中损伤

目的：急性缺血性脑卒中因其高致残率和死亡率而面临重大挑战。铁凋亡是由铁诱导的氧化应激引发的一种受调控的细胞死亡形式，在中风损伤中起关键作用。补阳还五汤治疗脑卒中的历史悠久，但其治疗铁下垂的机制尚不清楚。方法：网络药理学预测BHD的有效成分和作用途径，超高效液相色谱-串联质谱法（UPLC-MS/MS）确定其主要成分。C57小鼠诱导大脑中动脉闭塞（MCAO），通过Longa评分和TTC染色评估神经功能缺损和梗死面积。通过染色和电镜观察组织病理学和超微结构变化，并用试剂盒检测生化指标（MDA、GSH、SOD、Fe²+）。Western blotting和qPCR分析凋亡相关蛋白、Nrf2定位和基因表达。体外，在氧-葡萄糖剥夺/再氧化（OGD/R）条件下，评估HT22细胞活力和ROS水平。蛋白表达、Nrf2相互作用和核易位也被研究。结果：网络药理学显示BHD作用于脑梗死的关键通路，包括铁下垂和抗氧化通路。BHD能改善MCAO小鼠的神经功能，使梗死面积减少10% ~ 50%，显著降低氧化应激标志物（MDA、Fe2+）水平，提高抗氧化剂（GSH、SOD）活性。组织病理学和超微结构分析显示神经元损伤减轻，线粒体结构改善。Western blot和qPCR结果显示GPX4和Nrf2上调，Keap1下调，Nrf2核易位。体外应激条件下，BHD可提高HT22细胞活力，降低ROS。蛋白分析证实Nrf2、GPX4和HO-1增加，Keap1减少，Nrf2核易位增强。Nrf2抑制剂实验证实BHD的作用是Nrf2介导的。结论：在临床前研究中，BHD通过Nrf2/GPX4通路抑制铁下沉，在缺血性卒中中发挥神经保护作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Drug Design, Development and Therapy CHEMISTRY, MEDICINAL-PHARMACOLOGY & PHARMACY

CiteScore

9.00

自引率

0.00%

发文量

382

审稿时长

>12 weeks

期刊介绍： Drug Design, Development and Therapy is an international, peer-reviewed, open access journal that spans the spectrum of drug design, discovery and development through to clinical applications. The journal is characterized by the rapid reporting of high-quality original research, reviews, expert opinions, commentary and clinical studies in all therapeutic areas. Specific topics covered by the journal include: Drug target identification and validation Phenotypic screening and target deconvolution Biochemical analyses of drug targets and their pathways New methods or relevant applications in molecular/drug design and computer-aided drug discovery* Design, synthesis, and biological evaluation of novel biologically active compounds (including diagnostics or chemical probes) Structural or molecular biological studies elucidating molecular recognition processes Fragment-based drug discovery Pharmaceutical/red biotechnology Isolation, structural characterization, (bio)synthesis, bioengineering and pharmacological evaluation of natural products** Distribution, pharmacokinetics and metabolic transformations of drugs or biologically active compounds in drug development Drug delivery and formulation (design and characterization of dosage forms, release mechanisms and in vivo testing) Preclinical development studies Translational animal models Mechanisms of action and signalling pathways Toxicology Gene therapy, cell therapy and immunotherapy Personalized medicine and pharmacogenomics Clinical drug evaluation Patient safety and sustained use of medicines.