过表达脑红蛋白抑制双转基因AD小鼠脑中Aβ诱导的凋亡

发布时间：2019-01-09 10:11

【摘要】：目的:研究AD双转基因(APPswe/PS1d E9)小鼠侧脑室注射质粒p NGB后,过表达脑红蛋白(neuroglobin,NGB)对Aβ诱导的AD小鼠脑中细胞凋亡的影响及其潜在机制。方法:将24只鉴定后的13月龄AD双转基因阳性小鼠(雌雄各半)随机分为对照组、侧脑室注射生理盐水+pc DNA3.1(1 g/L)组和侧脑室注射pc DNA3.1(1 g/L)+p NGB组。采用免疫组化检测鼠脑Aβ1-42表达情况,TUNEL染色检测脑内细胞的凋亡情况;Western blot检测鼠脑内与凋亡密切相关的cleaved caspase-3、caspase-9以及PI3K、p-Akt、Akt的蛋白水平。结果:与对照组和注射生理盐水组比较,注射p NGB组小鼠脑内Aβ1-42的表达明显受到抑制(P0.01),TUNEL染色阳性细胞数也明显减少(P0.01);过表达NGB能够明显抑制脑组织内cleaved caspase-3和caspase-9的蛋白表达(P0.01),促进Akt磷酸化水平的增强(P0.01)。结论:p NGB过表达能够明显抑制Aβ的生成并抑制Aβ诱导的细胞凋亡,其机制可能与激活PI3K/Akt通路进而抑制与凋亡密切相关的cleaved caspase-3和caspase-9的表达有关。
[Abstract]:Aim: to study the effect of overexpression of brain erythrin (neuroglobin,NGB) on apoptosis of AD mice induced by A 尾 and its potential mechanism after injection of plasmid p NGB into the lateral ventricle of AD double transgenic (APPswe/PS1d E9) mice. Methods: Twenty-four 13-month-old AD double transgenic positive mice (male and female) were randomly divided into control group, intracerebroventricular injection of normal saline pc DNA3.1 (1 g / L) group and lateral ventricle injection of pc DNA3.1 (1 g / L) p NGB group). The expression of A 尾 1-42 in the brain was detected by immunohistochemistry, the apoptosis of brain cells was detected by TUNEL staining, and the protein levels of cleaved caspase-3,caspase-9 and PI3K,p-Akt,Akt, which were closely related to apoptosis, were detected by; Western blot. Results: compared with the control group and the saline group, the expression of A 尾 1-42 in the brain of the mice injected with p NGB was significantly inhibited (P0.01), TUNEL staining positive cells were also significantly decreased (P0.01). Overexpression of NGB could significantly inhibit the expression of cleaved caspase-3 and caspase-9 protein in brain tissue (P0.01) and enhance the level of Akt phosphorylation (P0.01). Conclusion the overexpression of: p NGB can significantly inhibit the formation of A 尾 and the apoptosis induced by A 尾. The mechanism may be related to the activation of the PI3K/Akt pathway and the inhibition of the expression of cleaved caspase-3 and caspase-9, which are closely related to apoptosis.
【作者单位】：重庆医科大学神经科学研究中心病理教研室;
【基金】：国家自然科学基金青年科学基金资助项目(No.81100948)
【分类号】：R749.16

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