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为了检测α-亚麻酸(α-linolenic acid,ALA)和花生四烯酸(arachidonic acid,AA)对于人肝癌细胞系(HepG2细胞)中核转录因子Nrf 2和Ⅰ相代谢酶胆固醇7α-羟化酶(CYP7A1)的mRNA和蛋白质表达的影响,并探究CYP7A1是否受Nrf 2的调控,试验以不同浓度的ALA和AA诱导HepG2细胞24 h,之后采用Real-time PCR法和Western-blot法分别检测HepG2细胞内Nrf 2和CYP7A1的mRNA和蛋白质的表达量。结果表明:当使ALA浓度为0.25,0.5,1 mmol/L作用于HepG2细胞时,Nrf 2和CYP7A1的mRNA和蛋白质的表达量相比于细胞对照均呈剂量依赖性升高(P<0.01);当使AA浓度为0.25,0.5,1 mmol/L作用于HepG2细胞时,Nrf 2的mRNA和蛋白质的表达量相比于细胞对照呈剂量依赖性升高(P<0.01),但CYP7A1的mRNA和蛋白质的表达量相比于细胞对照则呈剂量依赖性减少(P<0.01)。说明不同剂量的ALA和AA对Nrf 2和CYP7A1的mRNA和蛋白质的表达量影响不同,Nrf 2和CYP7A1呈正相关或负相关。
In order to investigate the effects of α-linolenic acid (ALA) and arachidonic acid (AA) on nuclear transcription factor Nrf 2 and phase I metabolic enzymes in human hepatocellular carcinoma cell line HepG2 cells, cholesterol 7α-hydroxylase (CYP7A1) mRNA and protein expression, and explore whether CYP7A1 is regulated by Nrf2, HepG2 cells were induced by different concentrations of ALA and AA for 24 h, then Real-time PCR and Western-blot were used to detect HepG2 MRNA and protein expression of Nrf 2 and CYP7A1 in cells. The results showed that the mRNA and protein expressions of Nrf 2 and CYP7A1 in HepG2 cells were increased in a dose-dependent manner (P <0.01) compared with the control cells when the concentration of ALA was 0.25, 0.5 and 1 mmol / ; When the concentration of AA was 0.25, 0.5 and 1 mmol / L, the mRNA and protein expression of Nrf 2 in HepG2 cells increased in a dose-dependent manner (P <0.01), but the mRNA expression of CYP7A1 And protein expression in a dose-dependent manner compared to the cell control (P <0.01). The results showed that different doses of ALA and AA had different effects on mRNA and protein expression of Nrf 2 and CYP7A1, and there was a positive or negative correlation between Nrf 2 and CYP7A1.