背景:非酒精性脂肪性肝病(NAFLD)是肝脏相关发病率和死亡率的主要原因,肝脏脂肪变性是标志性症状。丹参(Smil,Dan-Shen)和纹状体DC(Lstr,川雄)常用于治疗心血管疾病,具有调节脂质代谢的潜能。然而,Smil/Lstr组合是否可用于治疗NAFLD及其调脂特性的潜在机制尚不清楚.
目的:评估短期高脂饮食(HFD)诱导的斑马鱼模型评估肝脏脂肪变性表型的可行性和可靠性,并研究Smil/Lstr的肝脏降脂作用。以及它的活性成分。
方法:在HFD斑马鱼模型中使用荧光成像和组织化学检查肝脏和多个其他器官系统的表型改变。内源性评估Smil/Lstr组合的肝脏特异性降脂作用。进一步探索了斑马鱼的活性分子和功能机制,人类肝细胞,和仓鼠模型。
结果:在5天的HFD斑马鱼中,在血管和肝脏中检测到明显的脂质积累,正如用油红O和荧光脂质探针染色增加所证明的。在模型中观察到肝肥大,伴有大泡性脂肪变性。Smil/Lstr组合给药可有效恢复HFD斑马鱼的脂质分布并减轻肝肥大。在油酸刺激的肝细胞中,Smil/Lstr组合显着减少脂质积累和细胞损伤。随后,基于斑马鱼的表型筛选,天然的苯酞senkyunolideI(SEI)被确定为介导肝脏中Smil/Lstr组合的降脂活性的主要分子。此外,SEI上调脂质代谢调节剂PPARα的表达和下调的脂肪酸转位酶CD36,而PPARα拮抗剂充分阻断SEI对肝性脂肪变性的调节作用。最后,在仓鼠模型中进一步验证了SEI对肝脏脂质积累和PPARα信号传导的作用.
结论:我们提出了一种基于斑马鱼的肝脂肪变性调节剂筛选策略,并发现Smil/Lstr组合及其成分SEI对肝脏脂质积累和PPARα信号传导的调节作用,提示其作为NAFLD治疗新候选药物的潜在价值。
BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is the leading cause of liver-related morbidity and mortality, with hepatic steatosis being the hallmark symptom. Salvia miltiorrhiza Bunge (Smil, Dan-Shen) and Ligusticum striatum DC (Lstr, Chuan-Xiong) are commonly used to treat cardiovascular diseases and have the potential to regulate lipid metabolism. However, whether Smil/Lstr combo can be used to treat NAFLD and the mechanisms underlying its lipid-regulating properties remain unclear.
OBJECTIVE: To assess the feasibility and reliability of a short-term high-fat diet (HFD) induced zebrafish model for evaluating hepatic steatosis phenotype and to investigate the liver lipid-lowering effects of Smil/Lstr, as well as its active components.
METHODS: The phenotypic alterations of liver and multiple other organ systems were examined in the HFD zebrafish model using fluorescence imaging and histochemistry. The liver-specific lipid-lowering effects of Smil/Lstr combo were evaluated endogenously. The active molecules and functional mechanisms were further explored in zebrafish, human hepatocytes, and hamster models.
RESULTS: In 5-day HFD zebrafish, significant lipid accumulation was detected in the blood vessels and the liver, as evidenced by increased staining with Oil Red O and fluorescent lipid probes. Hepatic hypertrophy was observed in the model, along with macrovesicular steatosis. Smil/Lstr combo administration effectively restored the lipid profile and alleviated hepatic hypertrophy in the HFD zebrafish. In oleic-acid stimulated hepatocytes, Smil/Lstr combo markedly reduced lipid accumulation and cell damage. Subsequently, based on zebrafish phenotypic screening, the natural phthalide senkyunolide I (SEI) was identified as a major molecule mediating the lipid-lowering activities of Smil/Lstr combo in the liver. Moreover, SEI upregulated the expression of the lipid metabolism regulator
PPARα and downregulated fatty acid translocase CD36, while a
PPARα antagonist sufficiently blocked the regulatory effect of SEI on hepatic steatosis. Finally, the roles of SEI on hepatic lipid accumulation and
PPARα signaling were further verified in the hamster model.
CONCLUSIONS: We proposed a zebrafish-based screening strategy for modulators of hepatic steatosis and discovered the regulatory roles of Smil/Lstr combo and its component SEI on liver lipid accumulation and
PPARα signaling, suggesting their potential value as novel candidates for NAFLD treatment.