Qu Jialin, Chen Qiuyue, Wei Tianfu, Dou Ning, Shang Dong, Yuan Dan
Clinical Laboratory of Integrative Medicine, The First Affiliated Hospital of Dalian Medical University, Dalian, China.
Department of Traditional Chinese Medicine, Shenyang Pharmaceutical University, Shenyang, China.
Front Pharmacol. 2022 Aug 30;13:915535. doi: 10.3389/fphar.2022.915535. eCollection 2022.
Puerariae Flos, a representative homology plant of medicine and food for alcoholism, has a long history of clinical experience and remarkable curative effect in the treatment of alcoholic liver disease (ALD). However, its effective forms and hepatoprotective mechanisms remain unknown. In the present study, a strategy based on UPLC-QTOF MS combined with mass defect filtering technique was established for comprehensive mapping of the metabolic profile of PF in rat plasma, urine, bile, and feces after oral administration. Furthermore, the absorbed constituents into plasma and bile with a relatively high level were subjected to the network analysis, functional enrichment analysis, and molecular docking to clarify the potential mechanism. Finally, the therapeutic effect of PF on ALD and predicted mechanisms were further evaluated using a rat model of alcohol-induced liver injury and Western blot analysis. In total, 25 prototype components and 82 metabolites, including 93 flavonoids, 13 saponins, and one phenolic acid, were identified or tentatively characterized . In addition, glucuronidation, sulfation, methylation, hydroxylation, and reduction were observed as the major metabolic pathways of PF. The constructed compound-target-pathway network revealed that 11 absorbed constituents associated with the 16 relevant targets could be responsible for the protective activity of PF against ALD by regulating nine pathways attributable to glycolysis/gluconeogenesis, amino acid metabolism, and lipid regulation as well as inflammation and immune regulation. In addition, four active ingredients (6″-O-xylosyltectoridin, genistein-7-glucuronide-4'-sulfate, tectoridin-4'-sulfate, and 6″-O-xylosyltectoridin-4'-sulfate) as well as two target genes (MAO-A and PPAR-α) were screened and validated to play a crucial role with a good molecular docking score. The present results not only increase the understanding on the effective form and molecular mechanisms of PF-mediated protection against ALD but also promote better application of PF as a supplement food and herbal medicine for the treatment of ALD.
葛花是一种具有代表性的药食同源解酒植物，在治疗酒精性肝病（ALD）方面有着悠久的临床经验和显著疗效。然而，其有效成分和保肝机制尚不清楚。在本研究中，建立了一种基于超高效液相色谱-四极杆飞行时间质谱联用（UPLC-QTOF MS）结合质量亏损过滤技术的策略，用于全面描绘口服给药后葛根花（PF）在大鼠血浆、尿液、胆汁和粪便中的代谢谱。此外，对血浆和胆汁中吸收水平相对较高的成分进行网络分析、功能富集分析和分子对接，以阐明潜在机制。最后，使用酒精性肝损伤大鼠模型和蛋白质免疫印迹分析进一步评估PF对ALD的治疗效果和预测机制。共鉴定或初步表征了25种原型成分和82种代谢产物，包括93种黄酮类化合物、13种皂苷和1种酚酸。此外，观察到葡萄糖醛酸化、硫酸化、甲基化、羟基化和还原是PF的主要代谢途径。构建的化合物-靶点-途径网络显示，11种吸收成分与16个相关靶点相关，可能通过调节糖酵解/糖异生、氨基酸代谢、脂质调节以及炎症和免疫调节等9条途径，对PF抗ALD的保护活性起作用。此外，筛选并验证了4种活性成分（6″-O-木糖基大豆苷元、染料木素-7-葡萄糖醛酸-4'-硫酸酯、大豆苷元-4'-硫酸酯和6″-O-木糖基大豆苷元-4'-硫酸酯）以及2个靶基因（MAO-A和PPAR-α）在分子对接评分良好的情况下发挥关键作用。本研究结果不仅增加了对PF介导的抗ALD有效形式和分子机制的理解，也促进了PF作为补充食品和草药在ALD治疗中的更好应用。
