Sun Yize, Zhao Haibin, Wang Zheyi
Department of Traditional Chinese Medicine, Qilu Hospital (Qingdao), Shandong University, Qingdao, Shandong, China.
Department of Cardiology, Third Affiliate Hospital, Beijing University of Chinese Medicine, Beijing, China.
Front Pharmacol. 2025 Jan 28;16:1499418. doi: 10.3389/fphar.2025.1499418. eCollection 2025.
Patients with myocardial infarction (MI) have a high incidence of depression, which deteriorates the cardiac function and increases the risk of cardiovascular events. Shuangxinfang (Psycho-cardiology Formula, PCF) was proved to possess antidepressant and cardioprotective effects post MI. However, the compounds of PCF remain unidentified, and the pertinent mechanism is still not systematic. The purpose of this study is to determine the ingredients of PCF, further to probe the underlying mechanism for MI with depression.
The compounds of PCF were qualitatively identified by LC-MS/MS. The optimal dosage for lavage with the PCF solution in rats was determined to be 1 mL/100 g/day for a duration of 5 days. We also detected the PCF components migrating to blood in the control and model rats. Then the targets of PCF compounds were searched on Swiss target database, and the targets of depression and MI were predicted on TTD, OMIM, GeneCards, DrugBank and PharmGkb database. All the targets were intersected to construct the Protein-Protein Interaction (PPI) network on Metascape platform and the herb-compound-target (HCT) network on Cytoscape, to identify the hub targets. GO and KEGG pathway enrichment analysis were conducted on DAVID platform. Molecular docking was modeled on AutoDock Vina software.
There were 142 bioactive compounds from PCF acting on 270 targets in a synergistic way. And a total of seven components migrating to blood were identified, including Miltionone I, Neocryptotanshinone, Danshenxinkun A, Ferulic acid, Valerophenone, Vanillic acid and Senkyunolide D. Then SRC and MAPK3 were obtained as the hub proteins by degree value in PPI network, and P2RY12 was picked out as seed proteins ranked by scores from MCODES. Further analysis of biological process and signaling pathways also revealed the significance of ERK/MAPK. Statistical analyses (e.g., GO and KEGG pathway enrichment, PPI network analysis) demonstrated the significance of the identified targets and pathways ( < 0.05). Molecular docking results showed that the binding energies were all less than -5 kcal/mol. The stability of Neocryptotanshinone possessed the lowest binding energy to MAPK3.
We identified PCF's bioactive compounds and predicted its therapeutic mechanism for MI with depression using LC-MS/MS and bioinformatics. Key targets SRC, MAPK3, and seed protein P2RY12 were crucial for PCF's cardio-neuroprotective effects. Neocryptotanshinone showed the strongest binding to MAPK3, suggesting it as a pivotal active ingredient. These findings offer new insights and targets for future research on PCF.
心肌梗死（MI）患者抑郁症发病率较高，这会使心脏功能恶化并增加心血管事件风险。双心方（Psycho-cardiology Formula，PCF）已被证明在心肌梗死后具有抗抑郁和心脏保护作用。然而，PCF的化合物成分尚未明确，相关机制也仍不系统。本研究旨在确定PCF的成分，并进一步探究其对心肌梗死伴抑郁症的潜在作用机制。
采用液相色谱-串联质谱（LC-MS/MS）对PCF的化合物进行定性鉴定。确定大鼠灌胃PCF溶液的最佳剂量为1 mL/100 g/天，持续5天。我们还检测了对照组和模型组大鼠中迁移至血液中的PCF成分。然后在瑞士靶点数据库中搜索PCF化合物的靶点，并在TTD、OMIM、GeneCards、DrugBank和PharmGkb数据库中预测抑郁症和心肌梗死的靶点。将所有靶点进行交集分析，在Metascape平台上构建蛋白质-蛋白质相互作用（PPI）网络，在Cytoscape上构建草药-化合物-靶点（HCT）网络，以识别核心靶点。在DAVID平台上进行基因本体（GO）和京都基因与基因组百科全书（KEGG）通路富集分析。使用AutoDock Vina软件进行分子对接建模。
PCF中有142种生物活性化合物以协同方式作用于270个靶点。共鉴定出7种迁移至血液中的成分,包括丹参酮Ⅰ、隐丹参酮、丹参新醌甲、阿魏酸、戊苯酮、香草酸和党参炔苷D。然后，通过PPI网络中的度数值获得SRC和MAPK3作为核心蛋白，并通过MCODE评分选出P2RY12作为种子蛋白。对生物过程和信号通路的进一步分析也揭示了细胞外信号调节激酶/丝裂原活化蛋白激酶（ERK/MAPK）的重要性。统计分析（如GO和KEGG通路富集、PPI网络分析）证明了所鉴定靶点和通路的重要性（P<0.05）。分子对接结果表明，结合能均小于-5 kcal/mol。隐丹参酮对MAPK3的结合能最低，稳定性最好。
我们使用LC-MS/MS和生物信息学方法鉴定了PCF的生物活性化合物，并预测了其对心肌梗死伴抑郁症的治疗机制。关键靶点SRC、MAPK3和种子蛋白P2RY12对PCF的心脏神经保护作用至关重要。隐丹参酮与MAPK3的结合最强，表明它是关键活性成分。这些发现为PCF的未来研究提供了新的见解和靶点。
