热应激预处理对热应激小鼠以及体外培养的神经元的保护作用及机制

发布时间：2018-08-25 18:46

【摘要】：目的高温环境对人体生理功能的影响是广泛的,包括热代谢、能量代谢、水盐代谢、神经内分泌系统、心血管系统等;高温还能够诱导体外培养的皮层神经元发生凋亡。热应激预处理可以明显的提高机体抗耐热能力。本实验通过:1.观测不同的热应激预处理方式对小鼠行为学的影响,建立间歇性热应激预处理模型。2.观测热应激预处理对小鼠行为学的影响,揭示热应激预处理对小鼠脑体功能的影响。3.检测热应激预处理对急性热应激小鼠脑组织NOS活力、nNOS蛋白和基因表达以及行为学的影响。观测热应激预处理对体外培养的神经元细胞热应激后的细胞形态、突出长度以及细胞活性的影响,揭示热应激预处理拮抗急性热应激的机制。方法 1.成年健康昆明系雄性小鼠(体重20±2.0g)随机分为3组:正常对照组,间歇性20min热应激预处理组,间歇性30min热应激预处理组,每组30只。建立小鼠间歇性20min以及30min热应激预处理模型,预处理结束后0、6、12、24、36h分别取6只动物进行实验。采用穿梭和斜板实验测试小鼠学习记忆功能。2.成年健康昆明系雄性小鼠(体重20±2.0g)随机分为3组:正常对照组,间歇性热应激预处理组,连续性热应激预处理组,每组30只。复制小鼠间歇性和连续性热应激预处理模型,预处理结束后0、6、12、24、36h分别取6只动物进行实验。采用穿梭和斜板实验测试小鼠学习记忆功能。3.成年健康昆明系雄性小鼠(体重20±2.0g)随机分为4组:正常对照组,热应激组,间歇性热应激预处理组,连续性热应激预处理组,每组30只。复制小鼠间歇性和连续性热应激预处理模型以及急性热应激模型,热应激结束后0、6、12、24、36h每组分别取6只动物进行实验。采用穿梭和斜板实验测试小鼠学习记忆功能。并且在热应激结束后6h、12h、24h用NADPH-d组织化学、ABC免疫组化、RT-PCR测定小鼠脑组织NOS的活性、nNOS的蛋白和基因表达。4.建立大鼠大脑皮层神经元原代培养技术,在培养6天后观察热应激预处理对热应激后细胞生长情况、生长活力及突起长度的影响。
[Abstract]:Objective The effects of hyperthermia on human physiological function are extensive, including heat metabolism, energy metabolism, water and salt metabolism, neuroendocrine system, cardiovascular system, etc. Hyperthermia can also induce apoptosis of cultured cortical neurons in vitro. Heat stress pretreatment can significantly improve the body's heat resistance. This experiment was carried out: 1. Observation No. The effects of the same heat stress preconditioning on the behavior of mice were observed. 2. The effects of heat stress preconditioning on the behavior of mice were observed to reveal the effects of heat stress preconditioning on the brain and body functions of mice. 3. The activity of NOS, nNOS protein and gene table in the brain of mice with acute heat stress were detected. The effects of heat stress preconditioning on the morphology, length and activity of cultured neurons after heat stress were observed to reveal the mechanism of heat stress preconditioning antagonizing acute heat stress.
Methods 1. Adult healthy male Kunming mice (weighing 20.0 g) were randomly divided into three groups: normal control group, intermittent 20-minute heat stress preconditioning group and intermittent 30-minute heat stress preconditioning group, 30 mice in each group. The models of intermittent 20-minute and 30-minute heat stress preconditioning were established in mice. Six animals were selected at 0, 6, 12, 24 and 36 hours after the preconditioning. 2. Adult healthy male Kunming mice (weighing 20 + 2.0g) were randomly divided into three groups: normal control group, intermittent heat stress preconditioning group and continuous heat stress preconditioning group with 30 mice in each group. 6 mice were selected for the experiment at 24 and 36 hours respectively. The shuttle and slant plate experiments were used to test the learning and memory function of mice. 3. Adult healthy male Kunming mice (weighing 20.0 g) were randomly divided into 4 groups: normal control group, heat stress group, intermittent heat stress preconditioning group, continuous heat stress preconditioning group, 30 mice in each group. Sexual heat stress preconditioning model and acute heat stress model, after heat stress 0, 6, 12, 24, 36 hours each group of six animals were taken for experiment. Shuttle and slant plate experiments were used to test the learning and memory function of mice. Protein and gene expression of NOS. 4. To establish the primary culture technology of rat cerebral cortex neurons and observe the effect of heat stress preconditioning on cell growth, growth vigor and process length after 6 days of culture.
【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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