摘要
目的探讨特丁基对苯二酚(tertiary butylhydroquinone,tBHQ)对氯化镧所致原代培养大鼠大脑皮层神经元氧化损伤的拮抗作用。方法将出生24 h内清洁级Wistar大鼠的胎鼠大脑皮质神经元进行原代培养,采用神经元特异性烯醇化酶(NSE)免疫细胞荧光法进行神经元的纯度鉴定;将神经元分别暴露于终浓度为0(对照)、0.125、0.25、0.5mmol/L氯化镧的DMEM培养基中染毒24 h,同时,设40μmol/L tBHQ与相应浓度氯化镧干预组。采用二乙酰二氯荧光素(DCFH-DA)探针测定神经元内活性氧(ROS)的水平,采用Western blot法测定神经元内核因子NF-E2相关因子(nuclear factor erythroid2-related factor,Nrf2)、谷胱甘肽S转移酶(glutathione S transferase,GST)和超氧化物歧化酶2(superoxide dismutase 2,SOD2)蛋白的表达水平。结果与对照组比较,各浓度氯化镧染毒组原代培养大鼠神经元ROS水平均升高,差异有统计学意义(P<0.05);且随着氯化镧染毒浓度的升高,原代培养大鼠神经元内的ROS水平呈上升趋势。与相同浓度氯化镧组比较,0.25、0.5 mmol/L氯化镧+40μmol/L t BHQ干预组原代培养大鼠神经元的ROS水平均较低,差异有统计学意义(P<0.05)。与对照组相比,各浓度氯化镧组神经元内Nrf2、GST和SOD2蛋白的表达水平均降低,除0.125 mmol/L氯化镧染毒组外,差异均有统计学意义(P<0.05);且随着氯化镧染毒浓度的升高,原代培养大鼠神经元内Nrf2、GST和SOD2蛋白的表达水平均呈下降趋势。与相同浓度氯化镧组比较,0.125、0.25、0.5 mmol/L氯化镧+40μmol/L tBHQ干预组原代培养大鼠神经元的Nrf2、GST和SOD2蛋白的表达水平均增加,除0.25 mmol/L氯化镧+40μmol/L tBHQ干预组SOD2蛋白的表达水平外,差异有统计学意义(P<0.05)。结论氯化镧可使神经元内ROS水平增加,Nrf2,GST和SOD2的蛋白表达水平减少,造成神经元发生氧化损伤;而tBHQ干预可拮抗镧所致的ROS水平增加、Nrf2及抗氧化蛋白的表达降低,对�
Objective To explore antagonistic effects of tert-butyl hydroquinone(tBHQ)on oxidative damage of primary cerebral cortical neurons in rats induced by lanthanum chloride(LaCl3 ).Methods The newborn Wistar rats within 24 hours were collected to performed primary neurons culture.Immunofluorescence(NSE)was used to identify the cultured neurons.Neurons were exposed to 0(control),0.125,0.25 and 0.5 mmol/L LaCl3 in DMEM medium respectively,and also tBHQ(40μmol/L)+0.125mmol/L LaCl3 ,tBHQ(40μmol/L)+0.25 mmol/L LaCl3 and tBHQ(40μmol/L)+0.5 mmol/L LaCl3 respectively for 24 hours respectively.Diacetyl dichlorofluorescein(DCFH-DA)was used to detect ROS levels of neurons.Western blot was used to measure Nrf2,GST and SOD2 protein expression levels of neurons.Results Compared with the control group,ROS levels of primary neurons in 0.125,0.25 and 0.5 mmol/L LaCl3 groups increased significantly(P<0.05),with a dose dependent manner.Compared with the related LaCl3 exposure groups,ROS levels of primary neurons in 0.25 mmol/L LaCl3 +40μmol/L tBHQ group and 0.5 mmol/L LaCl3 +40μmol/L tBHQ group decreased significantly(P<0.05).Compared with the control group,Nrf2,GST and SOD2 protein expression levels in 0.25 and 0.5 mmol/L LaCl3 groups decreased significantly(P<0.05),except 0.125 mmol/L LaCl3 group,and Nrf2,GST and SOD2 protein expression levels gradually decreased with the increase of LaCl3 exposure doses.Compared with the related LaCl3 exposure groups,Nrf2,GST and SOD2 protein expression levels of primary neurons in 0.125mmol/L LaCl3 +40μmol/L tBHQ group,0.25 mmol/L LaCl3 +40μmol/L tBHQ group and 0.5 mmol/L LaCl3 +40μmol/L tBHQ group increased significantly(P<0.05),except the expression level of SOD2 protein in 0.25 mmol/L LaCl3 +40μmol/L tBHQ group.Conclusion LaCl3 can cause increase of ROS level,decrease of Nrf2,GST and SOD2 protein expression,then lead to oxidative damage in neurons.tBHQ can antagonize the increase of ROS level,and the decrease of Nrf2,GST,SOD2 protein expression,which may play a protective effect on the
作者
张丽锦
李英綦
胡晓宇
孙亚玲
高祥
于淼
刘诗语
蔡原
ZHANG Li-jin;LI Ying-qi;HU Xiao-yu;SUN Ya-ling;GAO Xiang;YU Miao;LIU Shi-yu;CAI Yuan(School of Public Health China Medical University, Shenyang, Liaoning 110122,China)
出处
《环境与健康杂志》
CAS
北大核心
2018年第12期1035-1039,F0003,共6页
Journal of Environment and Health
基金
国家自然科学基金(81273117
81673220)
