目的 运用网络药理学结合实验验证探讨雷公藤红素调控肠上皮细胞胆固醇代谢的潜在靶点及作用机制。方法 应用网络药理学筛选雷公藤红素调控肠上皮细胞胆固醇代谢的关键靶点和通路。运用Autodock Vina软件进行分子对接验证。通过构建体外肠上皮细胞高胆固醇模型，考察雷公藤红素对肠上皮细胞胆固醇代谢及其关键通路和靶点表达的影响。结果 共获得94个雷公藤红素靶点、15 415个肠道靶点、14 177个胆固醇代谢靶点。取交集得75个共有靶点。蛋白质-蛋白质相互作用（protein-protein interaction，PPI）网络筛选得到1个关键子网络。基因本体（gene ontology，GO）功能富集分析发现网络中的靶点与多种生物功能相关。基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）通路富集分析发现，雷公藤红素调控肠道胆固醇代谢涉及多条途径，其中肠道脂质代谢的关键通路（fat digestion and absorption）与PPI网络中的关键子网络之一密切相关。分子对接结果表明，雷公藤红素与关键网络中的核心蛋白肝X受体α（liver X receptor α，LXRα）具有良好的结合亲和力。体外实验表明，雷公藤红素显著降低高胆固醇环境下肠上皮细胞脂质堆积（P＜0.05、0.01、0.001），显著上调三磷酸腺苷结合盒转运蛋白G5和G8（adenosine triphosphate-binding cassette transporters G5 and G8，ABCG5/8）蛋白表达（P＜0.05、0.01），并下调NPC1样细胞内胆固醇转运蛋白1（NPC1 like intracellular cholesterol transporter 1，NPC1L1）表达（P＜0.05）。结论 雷公藤红素调控肠上皮细胞胆固醇代谢的作用机制可能与调节LXRα促进胆固醇流出、抑制胆固醇摄取密切相关。

Objective To explore the potential targets and mechanism of celastrol on regulating cholesterol metabolism in intestinal epithelial cells using network pharmacology combined with experimental validation. Methods Network pharmacology was employed to screen key targets and pathways of celastrol in regulating cholesterol metabolism in intestinal epithelial cells. Molecular docking was performed using Autodock Vina software for validation. An in vitro model of high cholesterol in intestinal epithelial cells was constructed to investigate the impact of celastrol on cholesterol metabolism, key pathways and target expression. Results A total of 94 targets for celastrol, 15 415 intestinal targets, and 14 177 cholesterol metabolism targets were obtained. The intersection resulted in 75 common targets. A key subnetwork was obtained through protein-protein interaction (PPI) network screening. Gene ontology (GO) function enrichment analysis revealed the involvement of the network targets in various biological functions. Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis results showed that celastrol regulated intestinal cholesterol metabolism through multiple pathways, with the key pathway of intestinal lipid metabolism (fat digestion and absorption) closely related to the key subnetwork in PPI network. Molecular docking results indicated a strong binding affinity between celastrol and the core protein liver X receptor α (LXRα) in the key network. In vitro experiments demonstrated that celastrol significantly reduced cholesterol accumulation in intestinal epithelial cells under high cholesterol conditions (P < 0.05, 0.01, 0.001), upregulated the protein expressions of adenosine triphosphate-binding cassette transporters G5 and G8 (ABCG5/8) (P < 0.05, 0.01), and downregulated NPC1 like intracellular cholesterol transporter 1 (NPC1L1) protein expression (P < 0.05). Conclusion The mechanism of celastrol in regulating cholesterol metabolism in intestinal epithelial cells may be closely related to the modulation of LXRα to promote cholesterol efflux and inhibit cholesterol uptake.

R285.5

湖南省自然科学基金资助项目（2024JJ5302）；湖南中医药大学自然科学基金资助项目（Z2023XJYB12）；湖南中医药大学大学生创新创业训练项目（X202310541014）；湖南中医药大学本科生科研创新基金资助项目（2023BKS178）