| 引用本文: |
郑清华, 刘一卓, 秦子镒, 邱璐, 郑相颖, 王显.基于网络药理学和细胞实验探讨络风宁2号方治疗心力衰竭的作用机制[J].湖南中医药大学学报,2025,45(12):2296-2306[点击复制] |
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| 基于网络药理学和细胞实验探讨络风宁2号方治疗心力衰竭的作用机制 |
| 郑清华,刘一卓,秦子镒,邱璐,郑相颖,王显 |
| (北京中医药大学东直门医院, 北京 100700;北京中医药大学东直门医院心内科, 北京 100700;北京中医药大学东直门医院心血管中心, 北京 100700;北京中医药大学心血管病研究院, 北京 100700) |
| 摘要: |
| 目的 探究络风宁2号方（LFN2）治疗心力衰竭的作用机制。方法 通过TCMSP和SwissTargetPrediction数据库搜索LFN2的活性成分及作用靶点，利用GeneCards和OMIM数据库收集心力衰竭相关疾病靶点；通过Venny 2.1.0在线平台确定LFN2与心力衰竭的交集靶点后导入STRING数据库构建蛋白质-蛋白质相互作用（PPI）网络，借助Cytoscape 3.9.1软件绘制“药物-成分-靶点”网络图，利用R语言，进行GO功能富集与KEGG通路富集分析。采用分子对接验证LFN2活性成分与核心靶点的结合能力。通过细胞实验对筛选cAMP/Drp1及细胞凋亡通路进行验证：将细胞分为对照组、模型组、LFN2组、Db-cAMP 组、LFN2+Db-cAMP组，分别检测各组ANP/BNPmRNA及cAMP、Drp1及细胞凋亡相关蛋白cleaved Caspase-3、Bcl-2、Bax的表达情况。结果 网络药理学筛选出157个活性成分及553个作用靶点。KEGG通路富集分析主要与Wnt信号通路、Rap1信号通路、MAPK信号通路、PPAR信号通路、cAMP信号通路、胰高血糖素信号通路、肾素分泌通路密切相关。PPI网络显示，AKT1、ALB、TNF等为核心靶点，分子对接结果显示，主要成分Ginsenoside Rh4、Biatractylolide、Danshenxinkun D和Tumulosic acid与核心靶点GAPDH、TNF、IL6、AKT1、ALB的结合能均＜-5.10 kJ/mol。细胞实验表明，与对照组相比，模型组ANP/BNP mRNA表达升高（P<0.01)，cAMP含量下降（P<0.05），Drp1荧光强度及蛋白增多（P<0.05），促凋亡蛋白cleaved Caspase-3、Bax显著升高（P<0.01），抗凋亡蛋白Bcl-2下降（P<0.05）；与模型组相比，LFN2组、Db-cAMP组、LFN2+cAMP组ANP/BNP mRNA含量下降（P<0.05），cAMP含量升高（P<0.05），Drp1荧光及蛋白降低（P<0.05），抗凋亡蛋白Bcl-2增多（P<0.05），且LFN2+Db-cAMP组改善程度优于LFN2组。结论 LFN2可能通过调控cAMP/Drp1信号通路，抑制心肌细胞凋亡，发挥抗心力衰竭的作用。 |
| 关键词: 心力衰竭 络风宁2号方 网络药理学 cAMP Drp1 细胞凋亡 |
| DOI：10.3969/j.issn.1674-070X.2025.12.008 |
| 投稿时间：2025-09-24 |
| 基金项目:北京市科研基地建设项目（2025-JYB-KYPT-05）；国家自然科学基金项目（82074263）。 |
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| Mechanism of action of Luofengning Formula No.2 in treating heart failure based on network pharmacology and cellular experiments |
| ZHENG Qinghua, LIU Yizhuo, QIN Ziyi, QIU Lu, ZHENG Xiangying, WANG Xian |
| (Dongzhimen Hospital, Beijing University of Chinese Medicine, Dongcheng, Beijing 100700, China;Department of Cardiology, Dongzhimen Hospital, Beijing University of Chinese Medicine, Dongcheng, Beijing 100700, China;Cardiovascular Center, Dongzhimen Hospital Affiliated to Beijing University of Chinese Medicine, Beijing 100700, China;Institute of Cardiovascular Diseases, Beijing University of Chinese Medicine, Beijing 100700, China) |
| Abstract: |
| Objective To explore the mechanism of action of Luofengning No.2 Formula (LFN2) in treating heart failure. Methods The active components and potential targets of LFN2 were identified using TCMSP and SwissTargetPrediction databases, while heart failure-related disease targets were collected from GeneCards and OMIM databases. Intersecting targets between LFN2 and heart failure were determined using Venny 2.1.0 online platform, then imported into STRING database to construct a protein-protein interaction (PPI) network. Cytoscape 3.9.1 software was used to visualize the "drug-component-target" network. R language was employed for GO functional enrichment analysis and KEGG pathway enrichment analysis. Molecular docking was performed to validate the binding affinity between the active components of LFN2 and the core targets. Cellular experiments were conducted to validate the cAMP/Drp1 and apoptosis pathways. Cells were divided into control, model, LFN2, Db-cAMP, and LFN2+Db-cAMP groups. The expression levels of ANP/BNP mRNA, cAMP, Drp1, and apoptosis-related proteins (cleaved Caspase-3, Bcl-2, and Bax) were measured in each group. Results Network pharmacology identified 157 active components and 553 target proteins for LFN2. KEGG pathway enrichment analysis revealed significant associations with the Wnt signaling pathway, Rap1 signaling pathway, MAPK signaling pathway, PPAR signaling pathway, cAMP signaling pathway, glucagon signaling pathway, and renin secretion pathway. PPI network analysis identified AKT1, ALB, and TNF as core targets. Molecular docking results showed that the main active components (Ginsenoside Rh4, Biatractylolide, Danshenxinkun D, and Tumulosic acid) exhibited strong binding affinities (binding energy<-5.10 kJ/mol) with the core targets (GAPDH, TNF, IL6, AKT1, and ALB). Cellular experiments demonstrated that compared with the control group, the model group showed increased ANP/BNP mRNA content (P<0.01), decreased cAMP content (P<0.05), enhanced Drp1 fluorescence intensity and protein level (P<0.05), significantly elevated levels of pro-apoptotic proteins cleaved Caspase-3 and Bax (P<0.01), and decreased levels of anti-apoptotic protein Bcl-2 (P<0.05). Compared with the model group, the LFN2, Db-cAMP, and LFN2+Db-cAMP groups exhibited reduced ANP/BNP mRNA content (P<0.05), increased cAMP content (P<0.05), reduced Drp1 fluorescence intensity and protein level (P<0.05), and increased Bcl-2 protein level (P<0.05). The improvement was more pronounced in the LFN2+Db-cAMP group than in the LFN2 group. Conclusion LFN2 may exert anti-heart failure effects by regulating the cAMP/Drp1 signaling pathway and inhibiting cardiomyocyte apoptosis. |
| Key words: heart failure Luofengning Formula No.2 network pharmacology cAMP Drp1 apoptosis |
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