基于Nrf2信号通路探讨异甘草酸镁对雷公藤甲素损伤L-02细胞中相关蛋白表达的影响

目的基于Nrf2-ARE信号通路探讨异甘草酸镁对雷公藤甲素损伤L-02细胞的保护作用及相关机制。方法体外培养正常人肝L-02细胞,通过免疫荧光技术检测异甘草酸镁(60μmol·L^-1)对雷公藤甲素(80nmol·L^-1)损伤L-02细胞中Nrf2核转移的影响;采用Western blotting测定细胞核中Nrf2及细胞中细胞色素P450 3A4(CYP3A4)、多药耐药相关蛋白2(multidrug resistance associated protein 2,MRP2)和胆盐输出泵(bile salt export pump,BSEP)的蛋白表达。结果雷公藤甲素对L-02细胞核中Nrf2具有诱导作用(P<0.01),并促进Nrf2核内转移;异甘草酸镁预处理组较雷公藤甲素组降低L-02细胞核中Nrf2的表达(P<0.01);异甘草酸镁预处理后对雷公藤甲素损伤L-02细胞中CYP3A4、MRP2和BSEP蛋白表达具有诱导作用(P<0.01)。结论异甘草酸镁可减轻雷公藤甲素对L-02细胞造成的损伤作用,其机制可能与诱导细胞核中Nrf2以及细胞中CYP3A4、MRP2和BSEP受体有关。

Effect of Magnesium Isoglycyrrhizinate on Expression of Related Proteins in Triptolide-injured L-02 Cells Based on Nrf2 Signaling Pathway

OBJECTIVE To study on the protective effect of magnesium isoglycyrrhizinate on L-02 cells injured by triptolide and its mechanism based on Nrf2-ARE signaling pathway.METHODS Human hepatocyte L-02 were cultured in vitro.Effect of magnesium isoglycyrrhizinate(60 μmol·L^-1) on nuclear translocation of nuclear factor-E2-related factor 2(Nrf2) in triptolide-induced(80 nmol·L^-1) L-02 cells by immunofluorescence;effect of magnesium isoglycyrrhizinate on the protein expression of Nrf2 in nuclear and cytochrome P450 3 A4(CYP3 A4),multidrug resistance associated protein 2(MRP2) and bile salt export pump(BSEP) in cell were determined by Western blotting.RESULTS Triptolide up-regulated the protein expression of Nrf2(P<0.01),and induced Nrf2 transfered to the triptolide-induced L-02 cell nuclear.Compared with triptolide group,magnesium isoglycyrrhizinate could obviously decrease the protein expression of Nrf2 in nuclear(P<0.01).Magnesium isoglycyrrhizinate up-regulated the protein expression of CYP3 A4,MRP2 and BSEP in triptolide-induced L-02 cell(P<0.01).CONCLUSION Magnesium isoglycyrrhizinate can alleviate the damage of L-02 cells induced by triptolide,which may be related to the induction of Nrf2 receptors in the nucleus,CYP3 A4,MRP2 and BSEP receptors in cells.