摘要
[目的]基于代谢组学技术探究补骨脂乙素(IBC)致人源肝细胞系HepaRG细胞(human hepatoma-derived progenitor cells)肝毒性的机制。[方法]通过MTT法检测IBC对HepaRG细胞活力的影响,计算半数抑制率IC50值。利用Triple TOFTM5600系统检测HepaRG细胞上清在IBC干预24 h后的代谢物,采用主成分分析、正交偏最小二乘法判别分析和聚类分析等多元统计方法筛选各给药组与对照组的差异代谢物,并对其进行代谢通路富集分析。[结果]随着IBC剂量的升高,HepaRG的细胞活力显著降低。30μmol/L时表现出较明显的细胞毒性(P<0.05);与对照组相比,当给药浓度达到24.53μmol/L时,细胞活力降低10%;在41.22μmol/L时,细胞活力降低90%。通过计算,得出其IC50为32.89μmol/L,表明IBC具有较强的肝细胞毒性作用。富集分析显示,IBC诱导HepaRG细胞损伤主要富集苯丙氨酸、酪氨酸和色氨酸生物合成,缬氨酸、亮氨酸和异亮氨酸生物合成及组氨酸代谢等途径(P<0.05)。[结论]补骨脂乙素能够显著抑制HepaRG细胞活性,并且通过影响苯丙氨酸、组氨酸、精氨酸、赖氨酸等细胞代谢物及苯丙氨酸、精氨酸、缬氨酸、亮氨酸和异亮氨酸生物合成代谢通路诱导肝细胞损伤。
[Objective]To investigate the mechanism of hepatotoxicity of human hepatoma-derived progenitor cells(HepaRG)induced by isobavachalcone(IBC)based on metabolomics technology.[Methods]The effect of IBC on HepaRG cell viability was detected by MTT assay,and the half maximal inhibitory concentration(IC50)value was calculated.The metabolites of HepaRG cell supernatants after 24 h of IBC intervention were detected using the Triple TOFTM 5600 system,and multivariate statistical methods,such as principal component analysis,orthogonal partial least squares discriminant analysis,and cluster analysis,were used to screen for differential metabolites between each dosing group and the control group.Metabolic pathway enrichment analysis was then performed on these differential metabolites.[Results]With the increase of IBC dosage,the cell viability of HepaRG significantly decreased.At 30μmol/L,significant cytotoxicity was observed(P<0.001);compared with the control group,when the drug concentration reached 24.53μmol/L,the cell viability decreased by 10%;at 41.22μmol/L,cell viability decreased by 90%.By calculation,its IC50 was determined to be 32.89μmol/L,indicating that IBC has strong hepatotoxic effects.Enrichment analysis showed that IBC-induced HepaRG cell damage was mainly enriched in pathways such as phenylalanine,tyrosine,and tryptophan biosynthesis,valine,leucine,and isoleucine biosynthesis,and histidine metabolism(P<0.001).[Conclusion]Isobavachalcone can significantly inhibit the activity of HepaRG cells and induce liver cell injury by affecting cellular metabolites such as phenylalanine,histidine,arginine,and lysine,as well as the biosynthetic metabolic pathways of phenylalanine,arginine,valine,leucine,and isoleucine.
作者
马舒悦
王爱习
马超
申蓉
董雨桐
袭恒豫
周昆
MA Shuyue;WANG Aixi;MA Chao;SHEN Rong;DONG Yutong;XI Hengyu;ZHOU Kun(Center of Drug Safety Evaluation,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;State Key Laboratory of Component-based Chinese Medicine,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China)
出处
《天津中医药大学学报》
2026年第2期159-166,共8页
Journal of Tianjin University of Traditional Chinese Medicine
基金
天津市教委科研计划项目(2024KJ014)。
关键词
补骨脂乙素
非靶向代谢组学
肝毒性
化学计量学
富集分析
isobavachalcone
untargeted metabolomics
hepatotoxicity
stoichiometry
enrichment analysis
