基于蛋白质组学技术探讨人参皂苷Rg3治疗肝细胞癌的作用机制

目的 应用蛋白质组学技术探索人参皂苷Rg3治疗肝细胞癌（HCC）的作用机制。方法 采用C57BL/6小鼠接种Hepa1-6细胞诱导皮下肝癌模型。当肿瘤体积达到100～150 mm3时,随机分为模型组、人参皂苷Rg3组、索拉非尼组,灌胃给药。连续给药14天后,处死小鼠并完整剥离瘤体,检测各组小鼠瘤体质量并计算抑瘤率;HE染色法观察各组小鼠肿瘤组织病理形态;蛋白质组测序分析各组的差异表达蛋白,并将显著差异表达蛋白对应基因进行基因功能注释（GO）、京都基因与基因组百科全书（KEGG）富集分析。结果 使用人参皂苷Rg3治疗的荷瘤鼠肿瘤质量明显低于模型组（P<0.05）。人参皂苷Rg3组的抑瘤率为65.3%,索拉菲尼组的抑瘤率为77.84%。病理结果显示,模型组癌细胞密度较大,生长旺盛,Rg3组癌细胞密度较小,存在区域坏死。索拉非尼组癌细胞密度小,大片区域坏死。蛋白质组学检测发现,人参皂苷Rg3组相较于模型组共有65个差异表达蛋白上调,24个差异表达蛋白下调。索拉非尼组相较于模型组共有59个差异表达蛋白上调,13个差异表达蛋白下调。GO分析发现,Rg3组对比模型组差异表达蛋白主要富集在生物过程中的补体和凝血级联反应、热生成、脂质代谢和细胞内信号转导的负调控。KEGG分析发现,Rg3组对比模型组差异表达蛋白主要富集在热生成、脂质代谢、肿瘤坏死因子产生的调节及肌肉的补体和凝血级联反应通路。结论 人参皂苷Rg3对HCC的抑制作用涉及产热、脂质代谢、肿瘤坏死因子产生的调节及肌肉的补体和凝血级联反应等过程。

Objective To explore the mechanism of ginsenoside Rg3 in treating hepatocellular carcinoma (HCC) using proteomics technology. Methods C57BL / 6 mice were inoculated with Hepa1-6 cells to induce subcutaneous HCC models. When tumor volume reached 100-150 mm3, the mice were randomly divided into model group, ginsenoside Rg3 group, and sorafenib group, and administered the corresponding drugs by gavage. After 14 days, the mice were euthanized, tumors were excised, and tumor weights were measured to calculate tumor inhibition rates. Histopathological morphology of tumor tissues was observed by HE staining. Proteomic sequencing was performed to identify differentially expressed proteins among groups, which were then annotated for GO and KEGG pathway enrichment analysis. Results Tumor weight in the ginsenoside Rg3-treated group was significantly lower than that in the model group (P<0. 05). The tumor inhibition rate was 65. 3% in the ginsenoside Rg3 group and 77. 84% in the sorafenib group. Pathological analysis showed higher cancer cell density and vigorous growth in the model group, while the ginsenoside Rg3 group had lower cancer cell density with areas of necrosis, and the sorafenib group had even lower cancer cell density with large areas of necrosis. Proteomic analysis revealed 65 upregulated and 24 downregulated differentially expressed proteins in the ginsenoside Rg3 group compared to the model group, and 59 upregulated and 13 downregulated proteins in the sorafenib group compared to the model group. GO analysis indicated that differentially expressed proteins in the ginsenoside Rg3 group were mainly enriched in biological processes such as complement and coagulation cascades, thermogenesis, lipid metabolism, and negative regulation of intracellular signal transduction. KEGG analysis showed that these proteins were primarily involves in thermogenesis, lipid metabolism, regulation of tumor necrosis factor production, and complement and coagulation cascades in muscle. Conclusion The inhibitory effect of ginsenoside Rg3 on HCC involves processes including thermogenesis, lipid metabolism, regulation of tumor necrosis factor production, and complement and coagulation cascade.