背景:药物性肝损伤(DILI)逐渐成为导致急性肝衰竭的全球性常见问题,尤其是对乙酰氨基酚(APAP)引起的急性肝损伤。芍药苷(PF)具有广泛的治疗作用,可以缓解多种肝病。然而,在目前的研究中,它们之间的关系仍然缺乏研究。
目的:本研究旨在探讨PF对APAP诱导的肝损伤的保护作用及其可能的分子机制。
方法:C57BL/6J雄性小鼠注射APAP建立DILI模型,连续给予PF治疗5天。为了阐明其药理作用,通过高通量和其他技术阐明了PF在APAP诱导的DILI中的分子机制。
结果:结果表明血清ALP水平,γ-GT,AST,TBIL,通过PF的预防作用,APAP小鼠的ALT降低。此外,PF明显减轻肝组织炎症和水肿。同时,TUNEL染色和相关凋亡因子的结果与转录组学的结果一致,提示PF通过调节MAPK信号抑制肝细胞凋亡。此外,PF还作用于活性氧(ROS)以调节氧化应激以恢复受损的线粒体。更重要的是,透射电镜显示PF处理后自噬体的产生,并且PF还下调mTOR,并在mRNA水平上调自噬标志物如ATG5,ATG7和BECN1的表达,在蛋白水平上调LC3,p62,ATG5和ATG7,这意味着PF发挥其作用的过程伴随着自噬的发生。此外,结合MAPK的分子动力学模拟和蛋白质印迹,结果表明p38是APAP上PF的直接靶标。具体来说,PF通过下调MAPK/mTOR信号激活自噬,这反过来减少了APAP伤害。
结论:芍药苷通过MAPK/mTOR信号通路激活自噬抑制氧化应激和凋亡,减轻肝损伤。一起来看,我们的发现阐明了芍药苷在DILI中的作用和机制,有望为芍药苷的开发提供新的治疗策略。
BACKGROUND: Drug-induced liver injury (DILI) is gradually becoming a common global problem that causes acute liver failure, especially in acute hepatic damage caused by acetaminophen (APAP). Paeoniflorin (PF) has a wide range of therapeutic effects to alleviate a variety of hepatic diseases. However, the relationship between them is still poorly investigated in current studies.
OBJECTIVE: This work aimed to explore the protective effects of PF on APAP-induced hepatic damage and researched the potential molecular mechanisms.
METHODS: C57BL/6J male mice were injected with APAP to establish DILI model and were given PF for five consecutive days for treatment. Aiming to clarify the pharmacological effects, the molecular mechanisms of PF in APAP-induced DILI was elucidated by high-throughput and other techniques.
RESULTS: The results demonstrated that serum levels of ALP, γ-GT, AST, TBIL, and ALT were decreased in APAP mice by the preventive effects of PF. Moreover, PF notably alleviated hepatic tissue inflammation and edema. Meanwhile, the results of TUNEL staining and related apoptotic factors coincided with the results of transcriptomics, suggesting that PF inhibited hepatocyte apoptosis by regulated MAPK signaling. Besides, PF also acted on reactive oxygen species (ROS) to regulate the oxidative stress for recovery the damaged mitochondria. More importantly, transmission electron microscopy showed the generation of autophagosomes after PF treatment, and PF was also downregulated mTOR and upregulated the expression of autophagy markers such as ATG5, ATG7, and BECN1 at the mRNA level and LC3, p62, ATG5, and ATG7 at the protein level, implying that the process by which PF exerted its effects was accompanied by the occurrence of autophagy. In addition, combinined with molecular dynamics simulations and western blotting of MAPK, the results suggested p38 as a direct target for PF on APAP. Specifically, PF-activated autophagy through the downregulation of MAPK/mTOR signaling, which in turn reduced APAP injury.
CONCLUSIONS: Paeoniflorin mitigated liver injury by activating autophagy to suppress oxidative stress and apoptosis via the MAPK/mTOR signaling pathway. Taken together, our findings elucidate the role and mechanism of paeoniflorin in DILI, which is expected to provide a new therapeutic strategy for the development of paeoniflorin.