铅对SH-SY5Y细胞的Nrf2转录活性及HO-1和γ-GCSc蛋白表达的影响
Effect of Lead on Transcriptional Activity of Nrf2 and Protein Expression of HO-1 and γ-GCSc in SH-SY5Y Cells
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摘要:
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、125 μmol/L)对体外培养的人神经母细胞瘤细胞(SH-SY5Y)染毒12、24 h 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。 Abstract:Objective To examine the nuclear factor erythroid 2-related factor 2 (Nrf2) transcriptional activity and the protein levels of heme oxygenase 1 (HO-1) and γ-glutamylcysteine synthetase (γ-GCSc) in SH-SY5Y cells induced by lead. Methods SH-SY5Y cells cultured in vitro were exposed to 0, 5, 25, and 125 μmol/L lead acetate solutions. After 12 h and 24 h of exposure, cell viability was determined by methly thiazolyl tetrazolium assay, and capacity of Nrf2-antioxidant response element (ARE) binding was measured by electrophoretic mobility shift assay. Western blot was used to detect the protein levels of HO-1 and γ-GCSc. Results Compared with the control group, the viability of the lead acetate cultured cells was significantly decreased; the capacity of Nrf2-ARE binding and the protein levels of HO-1 and γ-GCSc were obviously increased (P< 0.05). Conclusion Lead toxicity in SH-SY5Y cells presents in a dose-dependent manner. Lead can activate the transcriptional activity of Nrf2, and upregulate the expressions of Nrf2-mediated antioxidant enzyme HO-1 and γ-GCSc.
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