目的 基于网络药理学及实验验证探讨芪苈强心胶囊治疗射血分数保留型心衰（heart failure with preserved ejection fraction，HFpEF）的作用机制。方法 将44个明确鉴定的芪苈强心胶囊体内代谢物检识有效成分，利用Swisstargetprediction、pharmmapper平台预测化合物靶点，OMIM、DisGenet、GenCard数据库中检索HFpEF疾病靶点；取交集靶点，利用String数据库和Cytoscape 3.7.2软件进行蛋白质-蛋白质相互作用（protein-protein interaction，PPI）和拓扑分析；Metescape平台对共同靶点进行基因本体（gene ontology，GO）功能富集分析及京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）通路富集分析。构建醋酸脱氧皮质酮（deoxycorticosterone acetate，DOCA）盐敏感型HFpEF大鼠模型，给予沙库巴曲缬沙坦或芪苈强心胶囊进行干预，给药8周后，利用小动物超声成像系统、ELISA、免疫荧光、Western blotting、qRT-PCR方法进行药效学评估和环磷酸鸟苷（cyclic guanosinc monophosphate，cGMP）-蛋白激酶G1（protein kinase G1，PKG1）信号通路相关靶点验证。结果 芪苈强心胶囊有效成分和HFpEF分别筛选出38个相同作用靶点；GO功能及KEGG通路富集分析得出其在血液循环、循环系统、心脏收缩等生物过程，膜筏、轴突、细胞质区等细胞成分，α-葡萄糖苷酶活性、内肽酶活性、水解酶活性等分子功能上起作用，涉及cGMP-PKG信号通路、肾素分泌、钙信号通路等。体内实验证实芪苈强心能够提高HFpEF大鼠射血分数，降低室壁肥厚程度，缩短等容舒张时间（isovolumic relaxation time，IVRT），提升左室舒张期充盈速度比值（P＜0.05）；提高血清一氧化氮（nitric oxide，NO）、cGMP水平（P＜0.05）；增强心肌组织内皮型一氧化氮合成酶（endothelial nitric oxide synthase，eNOS）荧光表达（P＜0.05、0.01）；增加eNOS、PKG1、心肌肌浆网Ca²⁺-ATP酶（sarco/endoplasmic reticulum Ca²⁺-ATPase，SERCA）、可溶性鸟苷酸环化酶α（soluble guanylate cyclase α，sGCα）蛋白表达水平（P＜0.05、0.01），降低cGMP特异性磷酸二酯酶5A（cGMP-specific phosphodiesterase 5A，PDE5A）蛋白表达水平（P＜0.05）；减少心肌组织B型脑钠肽（B-type brain natriuretic peptide，BNP）、β-肌球蛋白重链（myosin heavy chain β，β-MHC）的mRNA表达（P＜0.05）。结论 芪苈强心胶囊可通过成分-多靶点-多环节发挥HFpEF的治疗作用，其机制与调节cGMP-PKG信号通路有关。

Objective To explore the mechanism of Qiliqiangxin Capsule (芪苈强心胶囊, QLQX) in treatment of heart failure with preserved ejection fraction (HFpEF) based on network pharmacology and experimental verification. Methods Forty-four clearly identified exogenous substances in QLQX were identified as active ingredients, and targets of compounds were predicted by Swisstarget prediction and Pharmmapper platforms, HFpEF disease targets were retrieved from OMIM, Disgenet and Gencard databases. Intersection targets were collected, String database and Cytoscape 3.7.2 software were uses to perform protein-protein interaction network analysis (PPI) and topology analysis. Metescape platform were used to perform gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis for common targets. A salt-sensitive HFpEF rat model with deoxycorticosterone acetate (DOCA) was constructed and given sacubitril-valsartan or QLQX for intervention. After eight weeks of administration, small animal ultrasound imaging system, ELISA, immunofluorescence, Western blotting, qRT-PCR methods were used for pharmacodynamic evaluation and cyclic guanosinc monophosphate (cGMP)-protein kinase G1 (PKG1) signaling pathway related target validation. Results Thirty-eight identical targets were screened out from active ingredients of QLQX and HFpEF; GO function and KEGG pathway enrichment analysis showed that they were involved in biological process (such as blood circulation, circulatory system, cardiac contraction), cellular components (such as membrane rafts, axons, cytoplasmic region) and molecular function (such as α-glucosidase activity, endopeptidase activity, hydrolase activity), involving cGMP-PKG signaling pathway, renin secretion, calcium signaling pathway. In vivo experiments confirmed that QLQX increased ejection fraction, reduced degree of ventricular wall hypertrophy, shortened isovolumic relaxation time (IVRT), and increased ratio of left ventricular diastolic filling velocity in HFpEF rats (P < 0.05), increased nitric oxide (NO) and cGMP levels in serum (P < 0.05), enhanced myocardial endothelial nitric oxide synthase (eNOS) fluorescence expression (P < 0.05, 0.01), increased eNOS, PKG1, cardiac sarcoplasmic reticulum Ca²⁺-ATPase (SERCA), soluble guanylate cyclase α (sGCα) protein expressions (P < 0.05, 0.01), decreased cGMP-specific phosphodiesterase 5A (PDE5A) protein expression level (P < 0.05), decreased B-type brain natriuretic peptide (BNP) and β-myosin heavy chain (β-MHC) mRNA expressions in myocardial tissue (P < 0.05). Conclusion QLQX can exert the therapeutic effect of HFpEF through component-multi-target-multi-link, and its mechanism is related to the regulation of cGMP-PKG signaling pathway.

国家重点研发计划"中医药现代化研究"重点专项资助项目(2017YFC1700501);河北省自然科学基金资助项目(H2019106059);河北省中医药管理局科研计划项目(Z2022020)