氟、砷对原代培养的大鼠海马神经细胞氧化应激及凋亡影响的实验观察
首发时间:2016-02-26
摘要:目的 本研究通过体外细胞实验,观察氟、砷单独及联合染毒对大鼠海马神经细胞氧化应激及细胞凋亡的影响。方法 采用2因素3水平的析因设计,将体外培养的海马神经细胞分为9组,分别为对照(细胞培养液)组、低剂量亚砷酸钠(2 μmol/ L)染毒组、高剂量亚砷酸钠(10 μmol/ L)染毒组、低剂量氟化钠(10 μmol/ L)染毒组、高剂量氟化钠(50 μmol/ L)染毒组、低剂量亚砷酸钠(2 μmol/ L)+低剂量氟化钠(10 μmol/ L)联合染毒组、低剂量亚砷酸钠(2 μmol/ L)+高剂量氟化钠(50 μmol/ L)联合染毒组、高剂量亚砷酸钠(10 μmol/ L)+低剂量氟化钠(10 μmol/ L)联合染毒组以及高剂量亚砷酸钠(10 μmol/ L)+高剂量氟化钠(50 μmol/ L)联合染毒组。采用MTT还原法检测细胞存活情况;采用微量酶标法测定细胞中谷胱苷肽(GSH)含量;采用硫代巴比妥法(TBA)测定谷胱苷肽过氧化物酶(GSH-Px)含量;采用WST-I法测定超氧化物歧化酶(SOD)活力;采用化学比色法测定丙二醛(MDA)含量及过氧化氢酶(CAT)活力;采用流式细胞仪检测细胞早期凋亡率。结果 与对照组比较,高剂量砷、高剂量氟、高剂量砷+低剂量氟以及高剂量砷+高剂量氟联合染毒组海马神经细胞MTT吸光度值明显降低(P<0.05);高剂量砷+低剂量氟联合组及高剂量砷+高剂量氟联合组CAT活力明显降低(P<0.05);高剂量砷染毒组、高剂量氟染毒组、低剂量砷+高剂量氟联合组、高剂量砷+低剂量氟联合组及高剂量砷+高剂量氟联合组细胞GSH含量明显下降,(P<0.05);高剂量氟染毒组、高剂量砷+低剂量氟联合组及高剂量砷+高剂量氟联合组细胞GSH-Px活力明显下降(P<0.05);高剂量砷染毒组、高剂量氟染毒组、低剂量砷+高剂量氟联合组、高剂量砷+低剂量氟联合组及高剂量砷+高剂量氟联合组细胞MDA含量明显升高(P<0.05);高剂量砷染毒组、高剂量氟染毒组、低剂量砷+高剂量氟联合组、高剂量砷+低剂量氟联合组及高剂量砷+高剂量氟联合组细胞凋亡率明显升高(P<0.05)。结论 氟、砷可导致大鼠海马神经细胞氧化应激并引起细胞凋亡率升高,这可能是氟、砷对动物学习记忆能力产生影响的重要原因。
关键词: 劳动卫生与环境卫生学 亚砷酸钠 氟化钠 氧化应激 凋亡
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Effects of Fluoride and Arsenic on Oxidative Atress and Apoptosis in Primary Hippocampal Nearons of rats in Culture
Abstract:Objective: To observe the dffects of fluoride and arsenic on oxidative stress and apoptosis in primary hippocampal neurons of rats. Methods: Primary hippocampal neurons of rats in culture were randomly divided into 9 groups, control group (cell culture), low dose NaAsO2 (2 μmol/L) group, high dose NaAsO2 (10 μmol/L) group, low dose NaF (10 μmol/L) group, high dose NaF (50 μmol/L) group, low dose NaAsO2 (2 μmol/L) and low dose NaF (10 μmol/L) combined group, low dose NaAsO2 (2 μmol/L) and high dose NaF (50 μmol/L) combined group, high dose NaAsO2 (10 μmol/L) and low dose NaF (10 μmol/L) combined group, high dose NaAsO2 (10 μmol/L) and high dose NaF (50 μmol/L) combined group. MTT test was used to detect survival rates of primary hippocampal neurons cells; contents of amounts of reduced glutathione (GSH) were detected with micro-ELISA; activity of catalase (CAT) was detected with visible light; activity of superoxide dismutase (SOD) was detected with WST-1; contents of malondialdehyde (MDA), activity of glutathione peroxidase (GSH-Px) were detected with colorimetry; the cell apoptosis rate was determined by flow cytometer. Results: Compared with the control group, MTT absorbance value of primary hippocampal neurons cells treated with high-dose NaAsO2 group, high-dose NaF group, high-dose NaAsO2 and low-dose NaF combined group, high-dose NaAsO2 and high-dose NaF combined group were significantly decreased (P<0.05); CAT contents of hippocampal neurons cells treated with high-dose NaAsO2 and low-dose NaF combined group, high-dose NaAsO2 and high-dose NaF combined group were significantly decreased (P<0.05); GSH contents of hippocampal neurons cells treated with high-dose NaAsO2 group, high-dose NaF group, low-dose NaAsO2 and high-dose NaF combined group, high-dose NaAsO2 and low-dose NaF combined group, high-dose NaAsO2 and high-dose NaF combined group were significantly decreased (P<0.05); GSH-Px contents of hippocampal neurons cells treated with high-dose NaF group, high-dose NaAsO2 and low-dose NaF combined group, high-dose NaAsO2 and high-dose NaF combined group were significantly decreased (P<0.05); MDA contents of hippocampal neurons cells treated with high-dose NaAsO2 group, high-dose NaF group, low-dose NaAsO2 and high-dose NaF combined group, high-dose NaAsO2 and low-dose NaF combined group, high-dose NaAsO2 and high-dose NaF combined group were significantly increased (P<0.05); the cell apoptosis rate in high-dose NaAsO2 group, high-dose NaF group, low-dose NaAsO2 and high-dose NaF combined group, high-dose NaAsO2 and low-dose NaF combined group, high-dose NaAsO2 and high-dose NaF combined group were significantly increased (P<0.05). Conclusions: Fluoride and arsenic exposure might cause oxidative stress and cell apoptosis rate increase in hippocampal neurons cells. These effcts might be related to damaged learning and memory of rats.
Keywords: Occupational and Environmental Health Sodium arsenite Sodium fluoride Oxidative stress Apoptosis
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