安宫牛黄丸（ANP）是临床多年用于治疗脑出血的中药配方。它由牛黄、麝香等成分组成。古籍记载，ANP的多种成分具有清热、解毒、镇静等功效，可有效治疗昏迷、中风等病症。然而，ANP 潜在作用的潜在机制仍在研究中。

ANP是一种广泛用于治疗脑出血（ICH）的中药。然而，治疗效果背后的确切机制仍然很大程度上难以捉摸。本研究旨在采用综合网络药理学方法和实验验证，揭示 ANP 在对抗 ICH 中的作用和药理学分子机制。

ANP和ICH的分子靶标是从各种数据库中获得的，然后使用STRING数据库构建蛋白质-蛋白质相互作用（PPI）网络。此外，分别使用 Metascape 数据库和 Cytoscape 进行基因本体（GO）富集和京都基因和基因组百科全书（KEGG）分析。最后进行分子对接。我们进行了一系列行为测试、免疫组织化学染色、TUNEL染色和Western Blot来验证ANP的效果。

IL-6、JUN、MMP9、IL-1β、VEGFA是主要候选靶点，与流体剪切应力、动脉粥样硬化、TNF信号通路等相关。提示ANP抗ICH的潜在机制可能主要与焦亡、炎症。体内验证表明，ANP治疗显着减少了TUNEL阳性细胞的数量，并且ANP抑制了Iba-1阳性神经元的激活，并抑制了炎症因子和细胞焦亡指标的表达。此外，ANP还提高了ICH小鼠的认知水平和运动能力。

该研究结果结合虚拟筛选和实验验证表明，ANP通过调节炎症和焦亡途径，在保护大脑免受神经元损伤方面做出了重要贡献，为未来的研究奠定了基础和创新思路。

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Ethnopharmacological relevance

Angong Niuhuang Pill (ANP) is a traditional Chinese medicine formula that has been used clinically for many years in the treatment of cerebral hemorrhage. It is composed of ingredients such as calculus bovis, moschus, and others. Ancient texts have documented that ANP's multiple components possess properties such as heat-clearing, detoxification, and sedation, which can be effective in treating conditions such as coma and stroke. However, the underlying mechanisms of ANP's potential actions are still under investigation.

ANP is a Chinese medicine widely utilized for the treatment of intracerebral hemorrhage (ICH). However, the precise mechanism underlying the therapeutic effects remains largely elusive. The present study aims to unravel the effects and pharmacological molecular mechanisms of ANP in combatting ICH, employing a comprehensive network pharmacology approach and experimental validation.

The molecular targets of ANP and ICH were obtained from various databases, followed by the construction of protein-protein interaction (PPI) networks using the STRING database. Further, gene ontology (GO) enrichment and Kyoto encyclopedia of genes and genomes (KEGG) analyses were conducted using the Metascape database and Cytoscape, respectively. Finally, molecular docking was performed. We performed a series of behavioral tests, immunohistochemical staining, TUNEL staining, and Western Blot to verify the effects of ANP.

IL-6, JUN, MMP9, IL-1β, VEGFA were the main candidate targets and were associated with fluid shear stress and atherosclerosis, TNF signaling pathway, etc. It is suggested that the potential mechanism of ANP against ICH may be mainly related to pyroptosis, inflammation. In vivo validation showed that ANP treatment significantly reduced the number of TUNEL-positive cells and ANP inhibited the activation of Iba-1 positive neurons, and suppressed the expression of inflammatory factors and pyroptosis indicators. In addition, ANP improved the cognitive level and motor ability of ICH mice.

The results of the study combined with virtual screening and experimental validation showed that ANP has an important contribution in protecting the brain from neuronal damage by regulating the pathways of inflammation and pyroptosis, laying the foundation and innovative ideas for future studies.