基于体外药效学结合分子对接技术研究穿山龙抗心肌缺血的作用机制

目的 通过液质联用技术、体外药效学及分子对接技术探究穿山龙抗心肌缺血的物质基础和作用机制。方法 利用UPLC-Q TOF-MS技术确定穿山龙醇提物中的主要成分，比较不同质量浓度（40、10、4、0.4、0.004 mg/mL）的穿山龙醇提物对H9c2心肌细胞缺氧/复氧（H/R）损伤后细胞活力的影响。应用AutoDock软件对潜在药效物质和关键靶点进行分子对接，相关结果采用Cytoscape 3.6.1进行构图和网络拓扑结构分析。结果 从穿山龙的醇提物中鉴定出7个化学成分，并证明穿山龙醇提物对H/R损伤H9c2心肌细胞具有一定的保护作用。筛选出5种（薯蓣皂苷元、薯蓣皂苷、对羟基苄基酒石酸、原薯蓣皂苷、薯蓣次皂苷A）与抗心肌缺血相关的活性成分，5种活性成分的分子对接结果显示潜在的药效物质与关键靶点内皮一氧化氮合酶（NOS）2、NOS3、环氧化酶/细胞色素C氧化酶多肽II（COX2）、热休克蛋白（HSP70）对接结果能量低于-8 kcal/mol。结论 穿山龙中的甾体皂苷类成分可能是其治疗心肌缺血的物质基础，穿山龙治疗心肌缺血是多成分与多靶点相互作用的结果。

Mechanism of Dioscoreae Nipponicae Rhizoma treating myocardial ischemia based on pharmacodynamics in vitro combined with molecular docking

Objective To study the material basis and mechanism of anti-myocardial ischemia of Dioscoreae Nipponicae Rhizoma through LC-MS technology, pharmacodynamics in vitro, and molecular docking technology. Methods The UPLC-Q TOF-MS technology was used to identify the main compounds of Dioscoreae Nipponicae Rhizoma. The effects of methanol extracts of Dioscoreae Nipponicae Rhizoma with different concentrations (40, 10, 4, 0.4, and 0.004 mg/mL) on the viability of H9c2 myocardial cells after hypoxia/reoxygenation injury were observed. Autodock vina software was used to perform molecular docking between potential pharmacodynamic substances and key targets. The related results were mapped and analyzed by Cytoscape 3.6.1. Results Seven chemical components were identified from the methanol extract of Dioscoreae Nipponicae Rhizoma, and the methanol extract of Dioscoreae Nipponicae Rhizoma was proved to have a protective effect on H9c2 myocardial cells injury by hypoxia/reoxygenation. Furthermore, the components of DNR were screened and five active components (diosgenin, dioscin, piscidic acid, protodioscin, and prosapogenin A) related to anti-myocardial ischemia were identified. The results of molecular docking showed that the energy of potential pharmacodynamic substances with key targets NSO2, NSO3, COX2, and HSP70 was lower than -8 kcal/mol. Conclusion The compounds of Dioscoreae Nipponicae Rhizoma such as steroid saponins might be the material basis for the treatment of myocardial ischemia. Dioscoreae Nipponicae Rhizoma treatment of myocardial ischemia is the result of the interaction between multi-component and multi-target.