目的探讨姜黄素对丙烯腈致大鼠星形胶质细胞毒性的保护作用及可能机制。方法原代培养大鼠大脑皮层星形胶质细胞,11 d后用2、5、10和20μmol/L姜黄素预处理6 h后再给予1.0 mmol/L丙烯腈染毒12 h。同时设立正常空白对照组和单纯丙烯腈(1.0 mmol/L)染毒组。以四甲基偶氮唑蓝(MTT)和LDH释放量为指标评定细胞活力和细胞毒性,免疫细胞荧光化学评价核因子E2相关因子2(Nrf2)核转位、蛋白印迹法测定Nrf2的表达及其下游II相解毒酶基因谷氨酸半胱氨酸合成酶(γ-GCS)和抗氧化酶血红素加氧酶(HO-1)的蛋白表达。结果与对照组比,丙烯腈染毒组细胞活力显著降低,LDH释放量显著增加。与丙烯腈染毒组比,10和20μmol/L姜黄素预处理可明显减少细胞活力的降低和LDH释放量的增加,且这种保护作用与增加Nrf2蛋白表达、促进Nrf2核转位以及诱导下游γ-GCS和HO-1的蛋白表达有关。结论在本试验条件下,姜黄素通过激活Nrf2-ARE信号通路保护丙烯腈所致大鼠星形胶质细胞损伤。
Objective To explore the protective effects of curcumin on neurotoxicity induced by acrylonitrile in astrocytes and potential role of activation of nuclear factor E2 related factor(Nrf2).Methods The postnatal rat cortical astrocytes were primarily cultured,and when all cells grew to confluence at about 11 days after seeding,various doses of curcumin(2 μmol/L,5 μmol/L,10 μmol/L,20 μmol/L) were pretreated for 6 hrs.Then cells were treated with 1.0 mM acrylonitrile(AN) for 12 h.The blank control group and AN group were also set up.The cell viability and cytotoxicity were assessed by MTT reduction and LDH leakage,respectively.The Nrf2 nuclear translocation was assessed by immunofluorescent staining and the expression of Nrf2,γ-GCS,and HO-1 was semiquantitatively detected by western blotting.Results AN significantly decreased cell viability and enhanced the LDH leakage.Compared to the AN group,the pretreatment of curcumin at concentrations of 10 μmol/L and 20 μmol/L significantly increased the cell viability,inhibited LDH leakage,stimulated the Nrf2 nuclear translocation,promoted Nrf2 expression and activated expression of the downstream γ-GCS and HO-1.Conclusion Curcumin exerted neuroprotection against AN-caused toxicity in astrocytes via activation of Nrf2-ARE pathway.