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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 investigate the protective effect of curcumin on rat astrocytes induced by acrylonitrile and its possible mechanism. Methods Rat cerebral cortical astrocytes were cultured for the first time. After 11 days, they were pretreated with 2, 5, 10 and 20 μmol/L curcumin for 6 h and then 1.0 mmol/L acrylonitrile was exposed for 12 h. At the same time, normal blank control group and acrylonitrile (1.0 mmol/L) exposure group were established. Cell viability and cytotoxicity were assessed using MTT and LDH release as indicators, and nuclear factorial nuclear factor 2 (Nrf2) nuclear translocation was detected by immunocytochemistry, and Nrf2 expression was measured by Western blotting. Its downstream phase II phase detoxification enzyme gene glutamic acid cysteine synthetase (γ-GCS) and antioxidant enzyme heme oxygenase (HO-1) protein expression. Results Compared with the control group, the cell viability of the acrylonitrile-treated group was significantly reduced, and the release of LDH was significantly increased. Compared with the acrylonitrile-treated group, pretreatment with 10 and 20 μmol/L curcumin significantly reduced cell viability and increased LDH release, and this protective effect increased Nrf2 protein expression, promoted Nrf2 nuclear translocation, and induced downstream γ-GCS and HO-1 protein expression. Conclusion Under the experimental conditions, curcumin protects rat astrocytes from damage induced by acrylonitrile through activation of Nrf2-ARE signaling pathway.