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目的探讨莱菔硫烷(sulforaphane,SFN)对人肝细胞系HHL-5细胞硫氧还蛋白还原酶-1(thioredoxin reductase1,TR1)的诱导作用及机制。方法采用WST试验观察SFN对HHL-5细胞24h的细胞毒作用,及TaqMan one-step RT-PCR测定对TR1mRNA的诱导,蛋白印迹法(Western blotting)测定核因子E2相关因子2(nuclear factor erythroid2-related factor2,Nrf2)的蛋白表达。结果5mol/LSFN处理HHL-5细胞24h促进细胞增殖,更高剂量表现生长抑制作用;SFN处理HHL-5细胞24h呈剂量依赖性诱导TR1转录,10mol/LSFN对TR1的诱导为对照组的4.6倍;20μmol/LSFN处理1h后明显诱导了胞浆和胞核Nrf2蛋白表达,并且这种诱导呈明显的时间依赖性。结论SFN可能通过诱导转录因子Nrf2介导的TR1转录,从而增强机体抗氧化防御系统机能。
Objective To investigate the induction and mechanism of sulforaphane (SFN) on human hepatocyte cell line HHL-5 induced by thioredoxin reductase 1 (TR1). Methods The cytotoxic effect of SFN on HHL-5 cells was observed by WST and the induction of TR1 mRNA by TaqMan one-step RT-PCR. The expression of nuclear factor erythroid2- related factor2, Nrf2) protein expression. Results HHL-5 cells were treated with 5mol / L LSFN for 24 h to promote cell proliferation and showed higher growth inhibitory effect at a higher dose. SFH treatment induced a dose-dependent induction of TR1 transcription in HHL-5 cells and TRI up-regulated by 10 mol / L LSFN for 4.6 h ; Nrf2 protein expression in cytoplasm and nucleus was obviously induced by 20μmol / LSFN for 1h, and the induction was obviously time-dependent. Conclusion SFN may enhance the function of antioxidant defense system by inducing Nrf2-mediated transcription of TR1.