基于网络药理学和体外实验探究枸杞子的抗结直肠癌活性

Activity of Lycii fructus against Colorectal Cancer Based on Network Pharmacology and in Vitro Experiments

摘要: 目的：利用网络药理学预测和体外实验验证，探讨枸杞子提取物的抗癌活性及作用机制。方法：利用中药系统药理学数据库与分析平台(TCMSP)获取枸杞子的活性成分以及对应的靶点；通过TTD数据库、Gene Card数据库和DisGeNET数据库筛选结直肠癌的治疗靶点；采用Venny2.1在线工具获取枸杞子与结直肠癌的共同靶点，并用STRING数据库进行PPI (Protein-Protein Interaction)网络分析；借助Metascape网站实现基因本体(GO)富集分析和KEGG通路富集分析；采用溶剂法提取，并采用真空冷冻法进行浓缩干燥；利用MTT实验检测枸杞子的抗癌活性。结果：获取枸杞子活性成分35个；潜在的抗癌靶点121个，其中基于PPI分析度值，最有潜力的5个靶点分别是TP53、HSP90AA1、AKT1、MAPK1、RELA；KEGG富集显示枸杞子活性成分主要通过PI3K-Akt信号通路、HIF-1信号通路、松弛素信号通路等发挥作用；MTT实验表明黑枸杞和红枸杞粗提物均具有抗结直肠癌的活性。结论：初步筛选并明确了枸杞子的活性成分及其抗癌活性，为枸杞子的抗癌作用机制研究提供一定的参考。

Abstract: Objective: To study the activity and the molecular mechanism of the active ingredients from Lycii fructus against colorectal cancer based on network pharmacology and in vitro experiments. Meth-ods: The active components and corresponding targets of Lycii fructus were predicted by TCMSP; screening colorectal cancer related targets by TTD database, Gene Card database, and DisGeNET database; the common targets of Lycii fructus and colorectal cancer were obtained using Venny2.1 online tool, and PPI (Protein-Protein Interaction) networks analysis was conducted using STRING database; Gene Ontology (GO) enrichment and KEGG pathway enrichment were analyzed by Metascape website; MTT assay was used to examine the anticancer activity of the extract from Lycii fructus against colorectal cancer. Results: 35 active ingredients of Lycii fructus were predicted, with 121 potential anti-cancer targets identified. The 5 most promising targets based on PPI analysis were TP53, HSP90AA1, AKT1, MAPK1, and RELA. KEGG enrichment showed that the active ingre-dients from Lycii fructus mainly functioned via PI3K-Akt signaling pathway, HIF-1 signaling path-way, and relaxin signaling pathway. MTT assay was used to examine the effect of the extract from Lycii fructus on the colorectal cancer cell viability and its antioxidant activity. Conclusion: Based on the network pharmacology and in vitro experiments, the possible antitumor targets of the active ingredients from Lycii fructus were screened and their antitumor activity was verified in vitro, providing a reference for the further study of the anticancer mechanism of Lycii fructus.