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[目的]研究铅对人神经母细胞瘤细胞株(SH-SY5Y)的细胞核因子E2相关因子2(Nrf2)转录活性以及下游血红素加氧酶1(HO-1)和谷氨酸半胱氨酸合成酶(γ-GCSc)表达的影响。[方法]用不同剂量的醋酸铅溶液(0、5、25、125μmol/L)对体外培养的人神经母细胞瘤细胞(SH-SY5Y)染毒12、24h后,用四甲基偶氮唑蓝法测细胞存活率;用凝胶迁移试验测细胞核内Nrf2-抗氧化反应元件(ARE)结合能力;用Western blot法检测染毒细胞内HO-1和γ-GCSc的蛋白水平。[结果]与对照组相比,醋酸铅染毒使SH-SY5Y的存活率降低,细胞核内Nrf2-ARE结合能力升高,Nrf2调控的下游基因HO-1和γ-GCSc的蛋白表达水平也明显升高,差异有统计学意义(P<0.05)。[结论]铅对SH-SY5Y细胞毒性作用具有剂量反应关系。铅可激活SH-SY5Y细胞Nrf2的转录活性,并导致Nrf2介导的抗氧化酶HO-1和γ-GCSc的表达水平升高。
[Objective] To investigate the transcriptional activity of lead on human nuclear neuron E2-related factor 2 (Nrf2) and downstream heme oxygenase 1 (HO-1) and glutamate cysteamine in human neuroblastoma cell line SH-SY5Y Effect of acid synthetase (γ-GCSc) expression. [Methods] Human neuroblastoma cells (SH-SY5Y) cultured in vitro were exposed to various concentrations of lead acetate solution (0, 5, 25 and 125μmol / L) for 12 and 24 hours. The cell viability was assayed by gel permeation assay. The binding capacity of Nrf2-antioxidant elements (ARE) in nuclei was measured by gel shift assay. The protein levels of HO-1 and γ-GCSc were detected by Western blot. [Results] Compared with the control group, the lead-bearing toxicity of lead decreased the survival rate of SH-SY5Y and the binding capacity of Nrf2-ARE in the nucleus, and the protein expression levels of HO-1 and γ-GCSc downstream of Nrf2 were also significantly increased High, the difference was statistically significant (P <0.05). [Conclusion] Lead has a dose-response relationship to the cytotoxicity of SH-SY5Y. Lead activated the transcriptional activity of Nrf2 in SH-SY5Y cells and resulted in the increase of Nrf2-mediated expression of antioxidant enzymes HO-1 and γ-GCSc.