牛磺酸锌对染汞大鼠神经元及学习记忆能力的保护作用

发布时间：2018-03-27 15:06

本文选题：汞　切入点：牛磺酸锌　出处：《天津医科大学》2006年硕士论文

【摘要】：目的汞是强神经毒物,牛磺酸和锌对脑发育有重要调节作用。本实验通过观测补充牛磺酸锌(TZC)对染Hg大鼠皮层和海马一氧化氮合酶(NOS)活力、神经元型一氧化氮合酶(nNOS)蛋白和基因表达的影响,以及观察在体外培养皮层神经元时,TZC对染Hg大鼠神经元生长活力及突起长度的影响,研究TZC拮抗Hg对神经系统损害的机理。方法选用雄性健康(出生后21d)Wistar大鼠60只,体重(2004±20)g,随机分为对照组、染汞组、汞低牛磺酸锌组和汞高牛磺酸锌组;分别饮用蒸馏水,4.3mg/kg·d HgCl_2饮水,4.3mg/kg·d HgCl_2饮水和0.23 g/L TZC饮水,4.3mg/kg·d HgCl_2饮水和0.46g/L TZC饮水,每组15只大鼠。喂养75天时,用Y-迷宫实验测试大鼠学习记忆能力;喂养90天时,用NADPH-d组织化学、nNOS免疫组织化学方法分别测定大鼠海马NOS活力、nNOS蛋白和基因表达。成年雄性健康昆明系小鼠20只,体重(20±2.0)g,随机分为对照组、小剂量氯化汞组、更小剂量氯化汞组和小剂量氯化汞+牛磺酸锌组;每组5只,隔天一次对小鼠进行腹腔注射HgCl_2(0.1mg/mL,0.05mg/mL),按10ml/kg体重注射:4小时后注射等剂量的TZC(100mg/mL)。10天后取小鼠脑组织,用RT-PCR测定nNOS的基因表达水平。通过建立皮层神经元原代培养实验技术,观察不同浓度TZC(0.25×10~-3mg/ml、0.5×10~-3mg/ml、1.0×10~-3mg/ml、2.0×10~-3mg/ml、4.0×10~-3mg/ml)和不同剂量HgCl_2(8×10~-6mg/ml、8×10~-5mg/ml、8×10~-4mg/ml、8×10~-3mg/ml、8×10~-2mg/ml)对培养皮层神经元生长活力及突起长度的影响。结果 1.行为学测试结果染汞75天后,与对照组大鼠相比,单纯染汞组大鼠Y-迷宫实验学会次数明显增多(P0.05)。与单纯染汞组相比,给予高TZC干预的染汞组大鼠Y-迷宫实验学会次数明显减少(P0.05)。
[Abstract]:Objective Mercury is a strong neurotoxin, taurine and zinc play an important role in regulating brain development. The activities of nitric oxide synthase (NOS) in cortex and hippocampus of rats exposed to Hg were observed by supplementation of TZC with zinc taurine. The effects of neuronal nitric oxide synthase (nNOS) protein and gene expression, and the effects of TZC on the growth activity and neurite length of neurons in cultured cortical neurons in vitro were observed, and the mechanism of TZC antagonistic effect of Hg on nervous system damage was studied. Methods 60 healthy male 21d)Wistar rats were randomly divided into control group, low taurine zinc group and high taurine zinc mercury group. Drinking distilled water 4.3mg / kg d HgCl_2 drinking water 4.3mg / kg d HgCl_2 drinking water and 0.23g / L TZC drinking water drinking water 4.3mg / kg d HgCl_2 drinking water and 0.46g/L TZC drinking water respectively. After 75 days of feeding, the learning and memory abilities of rats were tested by Ymaze test, and 90 days after feeding, NADPH-d histochemistry and nNOS immunohistochemical method were used to detect the activity of NOS protein and gene expression in hippocampus of rats. Twenty adult male healthy Kunming mice were randomly divided into control group, low dose mercuric chloride group, lower dose mercuric chloride group and small dose mercuric acid taurine zinc group, with 5 rats in each group. Mice were injected intraperitoneally with 0.1 mg / mL HgCl _ 2C _ (0.1) mg 路m ~ (-1) 路L ~ (-1) and 0.05 mg 路m ~ (-1) 路mL ~ (-1) by intraperitoneal injection the next day. The brain tissues of mice were taken after injection of the same dose of TZC(100mg/mL).10 4 hours later according to the weight of 10ml/kg, and the expression of nNOS gene was determined by RT-PCR. By establishing the primary culture technique of cortical neurons, the effects of different concentrations of TZC(0.25 脳 10 ~ (-3) mg 路ml ~ (-1) ~ (0.5 脳 10 ~ (-3) mg 路ml ~ (-1)) 1.0 脳 10 ~ (-3) mg 路ml ~ (-1) 路ml ~ (-1) ~ (2) 脳 10 ~ (-3) mg 路ml ~ (-1) and different doses of HgCl_2(8 脳 10 ~ (-3) ~ (-3) mg 路ml ~ (-1) ~ (8 脳 10 ~ (-10) ~ (-3) mg 路ml ~ (-1) ~ (-1)) on the growth activity and the length of neuronal protuberance were observed. Results. 1. After 75 days of behavior test, compared with the control group, the number of Y- labyrinth experimental societies of rats exposed to mercury alone increased significantly (P 0.05), and compared with that of the control group, there was a significant increase in the number of Y- labyrinth experimental societies of rats exposed to mercury alone. The number of Y- labyrinth experiment in mercury exposed group treated with high TZC significantly decreased P0. 05.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R363

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