Table_1_Interpreting the Pharmacological Mechanisms of Huachansu Capsules on Hepatocellular Carcinoma Through Combining Network Pharmacology and Experimental Evaluation.xlsx

Hepatocellular carcinoma (HCC) is one of the most fatal cancers across the world. Chinese medicine has been used as adjunctive or complementary therapy for the management of HCC. Huachansu belongs to a class of toxic steroids isolated from toad venom that has important anti-cancer property. This study was aimed to identify the bioactive constituents and molecular targets of Huachansu capsules (HCSCs) for treating HCC using network pharmacology analysis and experimental assays. The major bioactive components of HCSCs were determined using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). A series of network pharmacology methods including target prediction, pathway identification, and network establishment were applied to identify the modes of action of HCSCs against HCC. Furthermore, a series of experiments, including MTT, clonogenic assay, 3-D transwell, wound healing assay, as well as flow cytometry, were conducted to verify the inhibitory ability of HCSCs on HCC in vitro. The results showed that 11 chemical components were identified from HCSCs. The network pharmacological analysis showed that there were 82 related anti-HCC targets and 14 potential pathways for these 11 components. Moreover, experimental assays confirmed the inhibitory effects of HCSCs against HCC in vitro. Taken together, our study revealed the synergistic effects of HCSCs on a systematic level, and suggested that HCSCs exhibited anti-HCC effects in a multi-component, multi-target, and multi-pathway manner.

肝细胞癌（hepatocellular carcinoma, HCC）是全球致死率最高的恶性肿瘤之一。中医药（Chinese medicine）已被用作肝细胞癌诊疗的辅助或补充治疗手段。华蟾素是从蟾蜍毒液中分离得到的一类有毒甾体类化合物，具有显著的抗癌活性。本研究旨在通过网络药理学分析与实验验证，明确华蟾素胶囊（Huachansu capsules, HCSCs）治疗肝细胞癌的活性成分与分子靶点。研究采用超高效液相色谱-串联质谱（UPLC-MS/MS）技术鉴定了华蟾素胶囊的主要活性成分。随后通过靶点预测、通路富集、网络构建等一系列网络药理学方法，解析华蟾素胶囊抗肝细胞癌的作用机制。此外，本研究还开展了MTT实验、克隆形成实验、三维Transwell实验、划痕愈合实验以及流式细胞术等体外实验，验证华蟾素胶囊对肝细胞癌的体外抑制活性。实验结果表明，共从华蟾素胶囊中鉴定出11种化学成分。网络药理学分析显示，这11种成分共涉及82个抗肝细胞癌相关靶点以及14条潜在调控通路。体外实验进一步验证了华蟾素胶囊对肝细胞癌的抑制作用。综上，本研究从系统层面揭示了华蟾素胶囊的协同作用，并证实其通过多成分、多靶点、多通路的方式发挥抗肝细胞癌活性。