本文選題：氧化應(yīng)激 + 海馬　； 參考：《浙江大學(xué)》2008年碩士論文

【摘要】： 目的:研究大鼠N-Shc蛋白的表達(dá),以及經(jīng)H_2O_2刺激后不同時(shí)間點(diǎn)大鼠海馬N-Shc蛋白的磷酸化水平改變,和不同濃度H_2O_2刺激后大鼠海馬N-Shc蛋白與PKCδ蛋白的磷酸化水平以及兩者結(jié)合的改變,探討氧化應(yīng)激對大鼠海馬N-Shc蛋白功能的影響。方法:運(yùn)用免疫印跡技術(shù),檢測:①Shc家族蛋白在大鼠神經(jīng)系統(tǒng)及其它系統(tǒng)中的表達(dá);②N-Shc和PKCδ蛋白在不同月齡大鼠全腦中的表達(dá);③H_2O_2刺激后不同時(shí)間點(diǎn)大鼠海馬N-Shc蛋白的磷酸化水平改變;④不同濃度H_2O_2刺激后大鼠海馬N-Shc蛋白與PKCδ蛋白的磷酸化水平以及兩者結(jié)合的改變。所得數(shù)據(jù)以正常對照的單位密度值為標(biāo)準(zhǔn)進(jìn)行校正,用SPSS forwindows 13.0軟件進(jìn)行單因素方差分析。結(jié)果:大鼠N-Shc蛋白的表達(dá)集中在神經(jīng)系統(tǒng),在心臟、肝臟和肌肉等組織中幾乎沒有表達(dá);大腦N-Shc蛋白在孕12.5天和孕15.5天僅有少量表達(dá),在出生后表達(dá)量明顯增多,而PKCδ蛋白在大鼠大腦的各個(gè)發(fā)育時(shí)期都有表達(dá);H_2O_2刺激后,大鼠海馬N-Shc蛋白的磷酸化水平在5 min內(nèi)快速上升,并一直持續(xù)到30 min;不同濃度H_2O_2刺激后,N-Shc蛋白和PKCδ蛋白的磷酸化水平出現(xiàn)逐漸升高,H_2O_2濃度5mM組和對照組之間具有顯著性差異(P＜0.01);不同濃度H_2O_2刺激后,N-Shc蛋白和PKCδ蛋白的結(jié)合明顯增加,其結(jié)合量隨H_2O_2濃度的升高而增加,5mM H_2O_2刺激組和對照組之間具有顯著性差異(P＜0.05)。結(jié)論:大鼠海馬組織中的N-Shc蛋白,經(jīng)H_2O_2刺激后引起磷酸化改變,并存在刺激時(shí)間和刺激濃度的依從性;同時(shí),H_2O_2刺激可以促進(jìn)N-Shc蛋白和PKCδ蛋白的結(jié)合,并且二者的結(jié)合量隨H_2O_2濃度的升高而增加。提示,氧化應(yīng)激狀態(tài)下N-Shc蛋白功能的改變,有可能對抑制細(xì)胞凋亡具有積極的作用。
[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.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R363

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本文編號(hào)：1916038