摘要：目的：探讨蒙药古日古木－13中的有效成分对HepG2肝癌细胞的增殖迁移及其作用机制。方法：20只Wistar大鼠按照空白、高、中、低剂量药物分组每日一次进行灌胃，一周后心尖取血，制备含药血清完全培养基处理HepG2肝癌细胞，利用IncuCyteZoom动态细胞成像系统筛选含药血清作用最适浓度，选择最适浓度的含药血清作用于划痕处理的细胞，观察其迁移情况。通过网络药理学确定药物成分和疾病的共同蛋白，确定关键作用通路。蛋白免疫印迹检测Akt、p-Akt、mTOR、P-mTOR蛋白表达。结果：IncuCyte分析显示中浓度的含药血清对HepG2增值抑制最明显。IncuCyte显示细胞划痕后，中浓度含药血清处理的肝癌细胞迁移率下降（P＜0.05），抑制增殖。利用数据库筛选药物成分和疾病的共同靶点，Cytoscape平台分析出关键交叉蛋白是Akt，最后通过KEGG富集分析获得关联度较高的Akt相关信号通路。Western blot结果显示中浓度含药血清作用HepG2肝癌细胞48h后可以下调Akt、p-Akt、mTOR、P-mTOR蛋白表达量（P＜0.05)。结论：蒙药古日古木－13有效成分对HepG2肝癌细胞的增殖和迁移都有抑制作用，作用时间在48h开始明显，Akt是最关键的蛋白靶点。

Abstract:Objective: To investigate the active ingredients of Mongolian medicine Gurigumu-13 on the proliferation and migration of HepG2 hepatoma cells and its mechanism. METHODS: Twenty Wistar rats were divided into blank, high, medium and low doses of drugs and were given intragastric administration once a day. One week later, blood was collected from the apex of the heart. HepG2 liver cancer cells were prepared in complete medium containing serum, and screened by the IncuCyteZoom dynamic cell imaging system. The optimal concentration of drug-containing serum was used, and the optimal concentration of drug-containing serum was selected to act on the scratched cells to observe their migration. Through network pharmacology, the common proteins of drug components and diseases are identified, and the key action pathways are identified. The protein expressions of Akt, p-Akt, mTOR and P-mTOR were detected by western blot. Results: IncuCyte analysis showed that the medium concentration of drug-containing serum inhibited the proliferation of HepG2 the most. IncuCyte showed that after the cells were scratched, the migration rate of liver cancer cells treated with medium-concentration drug-containing serum decreased (P<0.05), and the proliferation was inhibited. Using the database to screen the common targets of drug components and diseases, the Cytoscape platform analyzes that the key crossover protein is Akt, and finally obtains the Akt-related signaling pathway with high correlation through KEGG enrichment analysis. Western blot results showed that the expression of Akt, p-Akt, mTOR and P-mTOR proteins could be down-regulated after the treatment of HepG2 liver cancer cells with medium concentration of drug-containing serum for 48 hours (P<0.05). Conclusion: The active ingredients of Mongolian medicine Gurigumu-13 have inhibitory effects on the proliferation and migration of HepG2 liver cancer cells, and the effect time is obvious at 48h. Akt is the most critical protein target.

内蒙古自然科学基金，（2020MS08108）2022年度自治区卫生健康科技计划项目（202201236）