氧化应激状态下大鼠海马N-Shc蛋白功能改变的探讨

发布时间：2018-05-20 20:03

本文选题：氧化应激 + 海马　；参考：《浙江大学》2008年硕士论文

【摘要】： 目的:研究大鼠N-Shc蛋白的表达,以及经H_2O_2刺激后不同时间点大鼠海马N-Shc蛋白的磷酸化水平改变,和不同浓度H_2O_2刺激后大鼠海马N-Shc蛋白与PKCδ蛋白的磷酸化水平以及两者结合的改变,探讨氧化应激对大鼠海马N-Shc蛋白功能的影响。方法:运用免疫印迹技术,检测:①Shc家族蛋白在大鼠神经系统及其它系统中的表达;②N-Shc和PKCδ蛋白在不同月龄大鼠全脑中的表达;③H_2O_2刺激后不同时间点大鼠海马N-Shc蛋白的磷酸化水平改变;④不同浓度H_2O_2刺激后大鼠海马N-Shc蛋白与PKCδ蛋白的磷酸化水平以及两者结合的改变。所得数据以正常对照的单位密度值为标准进行校正,用SPSS forwindows 13.0软件进行单因素方差分析。结果:大鼠N-Shc蛋白的表达集中在神经系统,在心脏、肝脏和肌肉等组织中几乎没有表达;大脑N-Shc蛋白在孕12.5天和孕15.5天仅有少量表达,在出生后表达量明显增多,而PKCδ蛋白在大鼠大脑的各个发育时期都有表达;H_2O_2刺激后,大鼠海马N-Shc蛋白的磷酸化水平在5 min内快速上升,并一直持续到30 min;不同浓度H_2O_2刺激后,N-Shc蛋白和PKCδ蛋白的磷酸化水平出现逐渐升高,H_2O_2浓度5mM组和对照组之间具有显著性差异(P＜0.01);不同浓度H_2O_2刺激后,N-Shc蛋白和PKCδ蛋白的结合明显增加,其结合量随H_2O_2浓度的升高而增加,5mM H_2O_2刺激组和对照组之间具有显著性差异(P＜0.05)。结论:大鼠海马组织中的N-Shc蛋白,经H_2O_2刺激后引起磷酸化改变,并存在刺激时间和刺激浓度的依从性;同时,H_2O_2刺激可以促进N-Shc蛋白和PKCδ蛋白的结合,并且二者的结合量随H_2O_2浓度的升高而增加。提示,氧化应激状态下N-Shc蛋白功能的改变,有可能对抑制细胞凋亡具有积极的作用。
[Abstract]:Objective: to study the expression of N-Shc protein and the phosphorylation of N-Shc protein in hippocampus of rats stimulated by H_2O_2 at different time points. The effect of oxidative stress on the function of N-Shc protein in hippocampus of rats was studied. The phosphorylation level of N-Shc protein and PKC 未 protein in hippocampus of rats stimulated by different concentrations of H_2O_2 and their binding changes were studied. Methods: Western blotting technique was used to detect the expression of PKC 未 protein and 2N-Shc protein in the nervous system and other systems of rats, and the expression of the protein 2N-Shc and PKC 未 protein in the whole brain of rats of different months of age. Changes of phosphorylation level of N-Shc protein in hippocampus of rats at different time points after 3H_2O_2 stimulation; changes of phosphorylation level and combination of N-Shc protein and PKC 未 protein in hippocampus of rats stimulated by different concentrations of H_2O_2. The data were calibrated according to the unit density of normal control and single factor analysis of variance (ANOVA) was carried out by SPSS forwindows 13.0 software. Results: the expression of N-Shc protein was concentrated in the nervous system, and hardly expressed in the heart, liver and muscle, while the expression of N-Shc protein in the brain was only small on the 12th day of gestation and 15.5 days of gestation, and the expression of N-Shc protein increased significantly after birth. However, after stimulation of PKC 未 protein at all developmental stages of rat brain, the phosphorylation level of N-Shc protein in hippocampus of rats increased rapidly within 5 min. The phosphorylation level of N-Shc protein and PKC 未 protein increased gradually after different concentrations of H_2O_2 stimulation. There was significant difference between 5mM group and control group (P < 0.01), and the binding of N-Shc protein and PKC 未 protein increased significantly after different concentrations of H_2O_2 stimulation. The binding capacity increased with the increase of H_2O_2 concentration. There was a significant difference between the 5 mm H_2O_2 stimulation group and the control group (P < 0.05). Conclusion: the phosphorylation of N-Shc protein in rat hippocampal tissue was induced by H_2O_2 stimulation, and there was compliance of stimulation time and stimulation concentration, and the stimulation of H _ S _ 2O _ 2 could promote the binding of N-Shc protein and PKC 未 protein. And the binding amount of the two increased with the increase of H_2O_2 concentration. The results suggest that the changes of N-Shc function under oxidative stress may play a positive role in inhibiting apoptosis.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R363

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