Sun Jianfang, Fan Yujie, Li Xiaoya, Qiu Yingxin, Lu Yuying, Shen Zhuo, Zhang Jinghai, Zhao Mingyi, Xu Yijia
School of Life Sciences and Biopharmaceutical Science, Shenyang Pharmaceutical University, Shenyang, Liaoning, 110016, China.
School of Medical Devices, Shenyang Pharmaceutical University, Shenyang, Liaoning, 110016, China.
Chem Biol Interact. 2025 Sep 5;418:111627. doi: 10.1016/j.cbi.2025.111627. Epub 2025 Jul 1.
Tripterygium wilfordii Hook. f. has been widely used in clinical practice due to its good anti-inflammatory and analgesic activities. However, the application is limited by some potential toxicity and side effects. Therefore, this study aimed to explore the potential heart risk components and potential mechanism of Tripterygium wilfordii. Traditional Chinese Medicine Database and Analysis Platform (TCMSP), Swiss Target Prediction, GeneCards and Open Target Platform databases were used to obtain the potential targets of Tripterygium wilfordii monomers and arrhythmia. GO pathway enrichment analysis was performed by Sangerbox. The potential interaction between monomers and hNav1.5 and hERG (two subtypes of ion channel protein) were predicted by AutoDock and verified by using whole cell patch clamp recordings. Intracellular calcium concentration of H9c2 myocardial cells were tested Fura2-AM fluorescence probe. Acute toxicity tests in mice were used to verify the potential cardiac risk in vivo through heart rate and representative cardiac enzyme profile. The results showed that 38 kinds of Tripterygium wilfordii components were screened by TCMSP, among them, 17 terpenoid monomer structures were acquired through PubChem database. 119 genes associated with disease and monomers were also obtained through various databases, and GO function analysis suggested that ion channels are probably target types of cardiac risk. The molecular docking results showed that 17 components could bind with hNav1.5 and hERG with different binding energy. Patch clamp results showed that mairin and wilforlide A could significantly inhibit the peak current of both hNav1.5and hERG and affect the dynamic property of both channels. Furthermore, mairin and wilforlide A could inhibit cell viability and increase intracellular calcium concentration of H9c2 myocardial cells, and mairin inhibited the heart rate ratio and increased the level of CK-MB. In conclusion, ion channel might be the potential cardiac risk target of Tripterygium wilfordii terpenoid, and mairin and wilforlide A might be main components of Tripterygium wilfordii causing cardiac risk.
雷公藤因其良好的抗炎和镇痛活性已在临床实践中广泛应用。然而，其应用受到一些潜在毒性和副作用的限制。因此，本研究旨在探索雷公藤潜在的心脏风险成分及潜在机制。利用中药数据库与分析平台（TCMSP）、瑞士靶点预测、基因卡片和开放靶点平台数据库获取雷公藤单体和心律失常的潜在靶点。通过Sangerbox进行GO通路富集分析。利用自动对接预测单体与hNav1.5和hERG（离子通道蛋白的两种亚型）之间的潜在相互作用，并通过全细胞膜片钳记录进行验证。使用Fura2-AM荧光探针检测H9c2心肌细胞的细胞内钙浓度。通过小鼠急性毒性试验，通过心率和代表性心肌酶谱验证体内潜在的心脏风险。结果表明，通过TCMSP筛选出38种雷公藤成分，其中通过PubChem数据库获得17种萜类单体结构。通过各种数据库还获得了119个与疾病和单体相关的基因，GO功能分析表明离子通道可能是心脏风险的靶标类型。分子对接结果表明，17种成分可与hNav1.5和hERG以不同结合能结合。膜片钳结果表明，美登素和雷公藤内酯醇A可显著抑制hNav1.5和hERG的峰值电流，并影响两种通道的动力学特性。此外，美登素和雷公藤内酯醇A可抑制H9c2心肌细胞的活力并增加细胞内钙浓度，美登素可抑制心率比值并增加CK-MB水平。总之，离子通道可能是雷公藤萜类潜在的心脏风险靶点，美登素和雷公藤内酯醇A可能是雷公藤导致心脏风险的主要成分。
