Ding Xin, Xu Min, Zhang Ya, Long Cuiping, Su Xuemei, Zhang Yang, Qiao Yan, Zhang Xingxing, Zhou Qian, Tan Guangguo, Ma Jing
Department of traditional Chinese medicine, Xijing Hospital, Air Force Medical University, Xi'an 710032, China; School of Pharmacy, Air Force Medical University, Xi'an 710032, China.
School of Pharmacy, Air Force Medical University, Xi'an 710032, China; The Third Stationed Outpatient Department, General Hospital of Central Theater Command, Wuhan 430070, China.
J Pharm Biomed Anal. 2025 Jan 15;253:116526. doi: 10.1016/j.jpba.2024.116526. Epub 2024 Oct 23.
Sini Decoction (SND) has been proven to be an effective formula to alleviate cardiac injury of myocardial infarction (MI). However, the potential mechanism of SND remains unclear. In this study, the MI rat model was established by ligating the left anterior descending coronary artery. A total of 17 SND-distributed components in heart were identified by using ultra-high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UHPLC-Q-TOFMS). The combination of four-dimensional (4D) proteomics and network pharmacology was employed to find the potential targets for therapeutic intervention, and molecular docking and cellular thermal shift assay (CETSA) were used to reveal the interactions between the potential targets and the potential active components distributed in heart of SND. 33 SND-effected proteins were identified by 4D proteomics, which was involved in carbon metabolism, fatty acid metabolism, valine, leucine and isoleucine degradation, tricarboxylic acid (TCA) cycle and PPAR signaling pathway. 17 potential SND-targeted direct proteins were screened by comparing SND-effected proteins generated from 4D proteomics with the MI-related proteins obtained from disease database. The potential relationships between 17 components and 17 potential SND-targeted direct proteins were established by molecular docking analysis, in which songorine, benzoylhypaconine, hypaconine, formononetin, and liquiritigenin could be bound to the surrounding amino acid residues in the binding pocket of Mtor, Parp1, Acadm, Crat, and Aldh2. Then, CETSA analysis further confirmed that songorine and benzoylhypaconine could increase the heat stability of Mtor and Parp1 in cardiac tissue lysate, respectively, which suggested that there existed direct interactions between songorine and Mtor, and benzoylhypaconine and Parp1. In summary, this work concluded that SND produced cardioprotective effects mainly through preserving energy metabolism, also demonstrated that the combination of 4D proteomics and network pharmacology was a promising tool for uncovering the molecular mechanisms of multi-components multi-targets effects of TCM.
四逆汤（SND）已被证明是减轻心肌梗死（MI）心脏损伤的有效方剂。然而，SND的潜在机制仍不清楚。在本研究中，通过结扎左冠状动脉前降支建立MI大鼠模型。使用超高效液相色谱-四极杆-飞行时间质谱（UHPLC-Q-TOFMS）鉴定了心脏中总共17种SND分布成分。采用四维（4D）蛋白质组学和网络药理学相结合的方法寻找治疗干预的潜在靶点，并利用分子对接和细胞热位移分析（CETSA）揭示潜在靶点与SND心脏分布的潜在活性成分之间的相互作用。通过4D蛋白质组学鉴定了33种受SND影响的蛋白质，这些蛋白质参与碳代谢、脂肪酸代谢、缬氨酸、亮氨酸和异亮氨酸降解、三羧酸（TCA）循环和PPAR信号通路。通过比较4D蛋白质组学产生的受SND影响的蛋白质与从疾病数据库获得的MI相关蛋白质，筛选出17种潜在的SND靶向直接蛋白质。通过分子对接分析建立了17种成分与17种潜在的SND靶向直接蛋白质之间的潜在关系，其中松雀碱、苯甲酰次乌头碱、次乌头碱、芒柄花素和甘草素可与Mtor、Parp1、Acadm、Crat和Aldh2结合口袋中的周围氨基酸残基结合。然后，CETSA分析进一步证实，松雀碱和苯甲酰次乌头碱可分别提高心脏组织裂解物中Mtor和Parp1的热稳定性，这表明松雀碱与Mtor、苯甲酰次乌头碱与Parp1之间存在直接相互作用。总之，本研究得出结论，SND主要通过维持能量代谢产生心脏保护作用，同时也证明了4D蛋白质组学和网络药理学相结合是揭示中药多成分多靶点作用分子机制的一种有前途的工具。
