大黄素改善D氨基半乳糖胺造成的L02细胞损伤

Emodin improves L02 cell damage caused by D-GalN

目的:探讨大黄素对急性肝细胞损伤的防护作用。方法:运用网络药理学筛选大黄素防护急性肝损伤的潜在靶点和通路。将L02细胞分为对照组、模型组、治疗组,对照组细胞予以普通培养液培养,模型组细胞培养液中加入浓度为40 mmol/L的D氨基半乳糖胺(D-GalN)干预12 h诱导急性肝细胞损伤模型,治疗组细胞在模型组的基础上预先加入10μmol/L的大黄素干预12 h。采用实时荧光定量PCR检测各组细胞SLC2A1、SLC2A4和Caspase-3基因表达情况,流式细胞术检测细胞凋亡,蛋白印迹法法检测Cleaved-Caspase-3和BCL2蛋白的表达。结果:网络药理学提示大黄素可以通过调节细胞凋亡信号通路,代谢相关信号通路干预急性肝衰竭。实验结果表明模型组肝细胞凋亡率均显著高于对照组(P<0.001);治疗组与模型组相比,肝细胞凋亡率显著降低(P<0.001);模型组细胞的SLC2A1、Caspase-3基因表达量上调(P<0.01),Cleaved-Caspase-3蛋白表达明显高于对照组;治疗组细胞的SLC2A1和Caspase-3基因表达量较模型组显著下调(P<0.05),Cleaved-Caspase-3蛋白表达明显低于模型组,BCL2蛋白表达显著高于模型组。结论:大黄素可能通过抗凋亡、抑制葡糖糖摄入等途径抑制D氨基半乳糖胺导致的肝损伤。

Objective:To investigate the protective effect of emodin on acute liver injury.Methods:Network pharmacology was used to screen the potential targets and pathways of emodin protection against acute liver injury.The L02 cells were divided into the control group,model group,and treatment group.The control group was cultured in ordinary medium,and the model group was treated with D galactosamine(D-GalN)with concentration of 40 mM for 12 h to induce acute liver injury model,and the treatment group was treated with 10 uM emodin for 12 h.Real-time fluorescence quantitative PCR was used to detect SLC2A1,SLC2A4,and Caspase-3 gene expression,cell apoptosis was detected by flow cytometry,and Cleaved Caspase-3 and BCL2 protein expression was detected by western blot.Results:The result of Network pharmacology suggested that emodin could intervene in acute liver failure by regulating the apoptosis signaling pathway and metabolism-related signaling pathway.The apoptosis rate of liver cells in the model group was significantly higher than that in the control group(P<0.001).Compared with the model group,the apoptosis rate of liver cells in the treatment group was significantly decreased(P<0.001).The expressions of SLC2A1 and Caspase-3 genes in the model group were up-regulated(P<0.01).The expression of SLC2A1 and Caspase-3 genes in the treatment group was significantly down-regulated compared with the model group(P<0.05),Cleaved Caspase-3 protein expression was significantly lower than the model group,and BCL2 protein expression was significantly higher than the model group.Conclusion:Emodin may inhibit liver injury through anti-apoptosis and inhibition of glucose intake.