Exploring the Therapeutic Mechanism of Xinbao Pill in Brain Injury After Cardiopulmonary Resuscitation Based on Network Pharmacology, Metabolomics, and Experimental Verification

IF 4.8 1区医学 Q1 NEUROSCIENCES

CNS Neuroscience & Therapeutics Pub Date : 2025-03-04 DOI:10.1111/cns.70297

Dongli Li, Qihui Wu, Zunjiang Li, Baijian Chen, Xing Sun, Qiqi Wu, Zhenzhu Ding, Linling Liu, Jiansong Fang, Ruifeng Zeng, Yong Gu, Banghan Ding

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引用次数: 0

Abstract

Background

Post-cardiopulmonary resuscitation brain injury (PBI) is essentially the cerebral ischemia reperfusion (CIR) injury, which is the main cause of death and long-term disability in patients with cardiac arrest. So far, there is no treatment for PBI; thus, it is urgent to develop new drugs or therapies for the prevention and treatment of brain injury after cardiopulmonary resuscitation. Although multiple constituent herbs or active ingredients of Xinbao Pill (XBP) have shown neuroprotective effects, whether XBP could play a therapeutic role on PBI is still unknown. This study aimed to illustrate the neuroprotective effect of XBP on PBI and probe the underlying mechanisms.

Method

We first performed the cell and animal experiments to validate the protective effect of XBP on neurological function. We next identified the potential differential metabolites via metabolomics analysis. We further conducted a comprehensive network pharmacology analysis including overlap gene analysis, protein–protein interaction network, and gene–biological process–module function network to preliminarily investigate the specific mechanism of action (MOA) of XBP against PBI. Finally, PCR, MTT, ELISA assay, as well as Western blotting experiments were made to validate our proposed molecular mechanisms.

Result

The in vitro experiment showed that XBP could increase cell viability and ameliorate cell morphological damage in PC12 cells exposed to oxygen–glucose deprivation and reoxygenation (OGD/RO) conditions. The in vivo experiment demonstrated that XBP improved the Neurologic Deficit Score (NDS), lowered the Neuron-Specific Enolase (NSE) level as well as reversed the typical neuropathological changes in PBI rats, indicating its neuroprotective effect on PBI. Further metabolomics analysis identified 94 differential metabolites after XBP treatment, and multiple metabolites were highly related to CIR. Moreover, network pharmacology results revealed that the therapeutic effect of XBP on PBI may be relevant to mitochondrial quality control (MQC). Mechanistically, XBP could not only promote the expressions of marker proteins including PGC1α, NRF1, TFAM, OPA1, MFN1 as well as MFN2 in mitochondrial biogenesis and mitochondrial fusion but also inhibit those proteins containing DRP1, MFF, FIS1, p62, PINK1, Parkin as well as LC3 in mitochondrial fission and mitophagy. Finally, AMP-activated protein kinase (AMPK) inhibitor was demonstrated to play a crucial role in regulating MQC.

Conclusions

Our study first determined that XBP might be an underlying anti-PBI formula, which also deciphered the potential MOAs of XBP against PBI by a network pharmacology approach combined with in vivo and in vitro experimental validation.

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基于网络药理学、代谢组学和实验验证探讨心保丸对心肺复苏后脑损伤的治疗机制

背景心肺复苏后脑损伤（PBI）本质上是脑缺血再灌注（CIR）损伤，是导致心脏骤停患者死亡和长期残疾的主要原因。到目前为止，还没有治疗PBI的方法；因此，迫切需要开发新的药物或治疗方法来预防和治疗心肺复苏后的脑损伤。虽然心保丸的多成分草药或有效成分已显示出神经保护作用，但心保丸是否对PBI有治疗作用尚不清楚。本研究旨在阐明XBP对PBI的神经保护作用并探讨其机制。方法首先通过细胞和动物实验验证XBP对神经功能的保护作用。接下来，我们通过代谢组学分析确定了潜在的差异代谢物。我们进一步通过重叠基因分析、蛋白-蛋白相互作用网络、基因-生物过程-模块功能网络等综合网络药理学分析，初步探讨XBP对PBI的特异性作用机制（MOA）。最后，通过PCR、MTT、ELISA和Western blotting实验验证了我们提出的分子机制。结果体外实验表明，XBP能提高缺氧-葡萄糖剥夺和再氧化（OGD/RO）条件下PC12细胞的细胞活力，改善细胞形态学损伤。体内实验表明，XBP可改善PBI大鼠神经功能缺损评分（NDS），降低神经元特异性烯醇化酶（NSE）水平，逆转PBI典型神经病理改变，提示XBP对PBI具有神经保护作用。进一步的代谢组学分析发现XBP治疗后存在94种差异代谢物，多种代谢物与CIR高度相关，网络药理学结果显示XBP对PBI的治疗作用可能与线粒体质量控制（MQC）有关。在机制上，XBP不仅能促进PGC1α、NRF1、TFAM、OPA1、MFN1、MFN2等标记蛋白在线粒体生物发生和线粒体融合中的表达，还能抑制DRP1、MFF、FIS1、p62、PINK1、Parkin、LC3等标记蛋白在线粒体分裂和线粒体自噬中的表达。最后，amp活化蛋白激酶（AMPK）抑制剂在调控MQC中发挥了至关重要的作用。我们的研究首先确定了XBP可能是一种潜在的抗PBI配方，并通过网络药理学方法结合体内和体外实验验证，破译了XBP抗PBI的潜在MOAs。

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

CNS Neuroscience & Therapeutics 医学-神经科学

CiteScore

7.30

自引率

12.70%

发文量

240

审稿时长

2 months

期刊介绍： CNS Neuroscience & Therapeutics provides a medium for rapid publication of original clinical, experimental, and translational research papers, timely reviews and reports of novel findings of therapeutic relevance to the central nervous system, as well as papers related to clinical pharmacology, drug development and novel methodologies for drug evaluation. The journal focuses on neurological and psychiatric diseases such as stroke, Parkinson’s disease, Alzheimer’s disease, depression, schizophrenia, epilepsy, and drug abuse.