Department of Pharmacy, The First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou, 450003, China; Henan Province Engineering Research Center for Clinical Application, Evaluation and Transformation of Traditional Chinese Medicine, Henan Provincial Key Laboratory for Clinical Pharmacy of Traditional Chinese Medicine, Henan Province Engineering Research Center of Safety Evaluation and Risk Management of Traditional Chinese Medicine, Zhengzhou, 450003, China.
School of Pharmacy, Henan University of Chinese Medicine, Zhengzhou, China; College Pharmaceutical Engineering of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, China.
J Ethnopharmacol. 2024 Dec 5;335:118600. doi: 10.1016/j.jep.2024.118600. Epub 2024 Jul 23.
Herb-induced liver injury (HILI) represents an exacerbated inflammatory response, with Psoraleae fructus (PF) and its preparations recently associated with hepatotoxicity. Licorice, historically recognized as a detoxifying herbal remedy, is considered to possess hepatoprotective properties. Our previous research identified bavachin, bakuchiol, and psoralidin (PSO) as potential toxic constituents in PF, while licochalcone B (LCB) and echinatin were identified as bioactive components in licorice. However, evidence regarding the interactions of active compounds in herbs and their underlying mechanisms remains limited.
The objective of this study is to assess the potential mechanisms through which LCB modulates immunological and anti-inflammatory responses to treat PSO-induced liver injury by using human hepatocyte cells (L02) and LPS-primed mice.
The ameliorative effects of LCB and echinatin on bavachin, bakuchiol, and PSO-induced liver injury were demonstrated in L02 cells. Subsequently, the efficacy of LCB on PSO-induced idiosyncratic liver injury was further validated in C57BL/6 mice under moderate inflammatory stress induced by LPS priming. The mechanisms were preliminarily explored with an integrated strategy of molecular docking, RT-PCR verification, and untargeted metabolomics.
The study shows that LCB significantly reduced cell injury induced by the three chemicals in PF and provided substantial protection against PSO-induced hepatic damage, as indicated by the levels of ALT, AST, and LDH. LCB normalized liver function and remarkedly alleviated hepatic lesions and inflammation caused by PSO in mice under moderate inflammatory stress. The mRNA profiles of both L02 cells and mice liver tissue revealed that LCB mitigated PSO-induced hepatotoxicity by regulating the gene expression of pro-inflammatory cytokines IL1B and TNF, as well as immunoinflammatory genes PIK3CA, AKT1, NFKB1, and NLRP3. Furthermore, untargeted metabolomics of liver tissue indicated that LCB could reverse the abnormal expression of 11 discriminatory metabolites, with the interrelationship between differential metabolites and target genes primarily clustering in glycerophospholipid metabolism, arachidonic acid metabolism, and phosphatidylinositol signaling system.
LCB demonstrated a superior anti-inflammatory and immunomodulatory effect on PSO-induced hepatotoxicity by modulating the inflammatory response and metabolic signaling system. Key interactive targets included phosphatidylcholine, phosphatidic acid, and subunit isoforms of PI3K.
草药诱导的肝损伤（HILI）代表了一种加剧的炎症反应，而补骨脂及其制剂最近与肝毒性有关。甘草，作为一种传统的解毒草药，被认为具有保肝作用。我们之前的研究发现补骨脂中的补骨脂素、异补骨脂素和花椒毒素（PSO）可能是有毒成分，而甘草中的甘草查尔酮 B（LCB）和茵陈色原酮是生物活性成分。然而，关于草药中活性化合物的相互作用及其潜在机制的证据仍然有限。
本研究旨在评估 LCB 通过调节免疫和抗炎反应来治疗 PSO 诱导的肝损伤的潜在机制，方法是使用人肝细胞（L02）和 LPS 预处理的小鼠。
在 L02 细胞中证明了 LCB 和茵陈色原酮对补骨脂素、异补骨脂素和 PSO 诱导的肝损伤的改善作用。随后，在 LPS 预处理诱导的中度炎症应激下，进一步验证了 LCB 对 C57BL/6 小鼠 PSO 诱导的特发性肝损伤的疗效。采用分子对接、RT-PCR 验证和非靶向代谢组学的综合策略初步探讨了机制。
研究表明，LCB 显著降低了 PF 中三种化学物质引起的细胞损伤，并为 PSO 诱导的肝损伤提供了显著的保护，这一点从 ALT、AST 和 LDH 的水平可以看出。LCB 使肝功能正常化，并显著减轻了 PSO 在中度炎症应激下对小鼠肝脏损伤和炎症的影响。L02 细胞和小鼠肝组织的 mRNA 谱表明，LCB 通过调节促炎细胞因子 IL1B 和 TNF 以及免疫炎症基因 PIK3CA、AKT1、NFKB1 和 NLRP3 的基因表达，减轻了 PSO 诱导的肝毒性。此外，肝组织的非靶向代谢组学表明，LCB 可以逆转 11 种差异表达代谢物的异常表达，差异代谢物与靶基因之间的相互关系主要聚集在甘油磷脂代谢、花生四烯酸代谢和磷脂酰肌醇信号系统中。
LCB 通过调节炎症反应和代谢信号系统，对 PSO 诱导的肝毒性表现出优越的抗炎和免疫调节作用。关键的交互靶点包括磷脂酰胆碱、磷脂酸和 PI3K 的亚基同工型。
