SAMP8小鼠海马内CD147与NCT的表达及其相互关系

发布时间：2018-04-04 22:10

本文选题：CD　切入点：NCT　出处：《神经解剖学杂志》2017年03期

【摘要】：目的:运用免疫荧光组织化学和免疫共沉淀技术(Co IP),探讨快速老化小鼠(SAMP8)海马组织中细胞外基质金属蛋白酶诱导因子(CD147)和单过性跨膜蛋白(NCT)间的表达情况,为寻找阿尔茨海默病(Alzheimer disease,AD)相关的治疗策略和方向提供理论依据。方法:3月龄雄性SAMP8小鼠6只。3只小鼠用于免疫荧光组织化学实验,经4%多聚甲醛灌注、取脑、石蜡包埋后,冠状面切片进行组织化学染色,观察CD147与NCT在海马区是否存在共定位的关系。3只用于Co IP实验,提取海马组织蛋白,与CD147抗体孵育后加入琼脂糖微珠A/G孵育过夜,离心后上清用于Western Blot实验检测CD147与NCT间的相互作用关系。结果:SAMP8海马区,CD147和NCT的阳性表达部位主要分布于CA1、CA3、CA4区的锥体细胞层和齿状回颗粒细胞层,二者在CA1、CA3区表达较强;标记部位为胞膜、胞浆,部分神经元在胞浆进核周处存在不连续的点、线状共标记部位。Co IP实验显示CD147与成熟型NCT相互作用,不与非成熟型NCT相互作用。结论:CD147和NCT在SAMP8小鼠海马区锥体细胞层表达较高,推测二者可能与海马的学习记忆功能密切相关;CD147与NCT存在相互作用的关系,推测CD147可能参与调节γ-分泌酶活性。
[Abstract]:Objective: to investigate the expression of extracellular matrix metalloproteinase-inducible factor CD147 (CD147) and single transmembrane protein (NCTs) in hippocampus of rapidly aging mice (SAMP8) by immunofluorescence histochemistry and co-immunoprecipitation techniques.To provide theoretical basis for finding the treatment strategy and direction related to Alzheimer disease (AD).Methods six male SAMP8 mice at the age of 3 months were used in immunofluorescence histochemistry experiment. The brains were taken from the brain and embedded in paraffin wax, and the sections were stained with histochemistry on the coronal sections after perfusion with 4% paraformaldehyde.The co-localization of CD147 and NCT in hippocampus was observed. 3 was used only for Co IP experiment to extract hippocampal tissue protein, incubate with CD147 antibody and incubate with agarose microbead A- / G overnight.The supernatant of centrifuge was used to detect the interaction between CD147 and NCT by Western Blot experiment.Results the positive expression sites of CD147 and NCT were mainly located in the pyramidal cell layer and dentate gyrus granulosa cell layer in CA1 / CA3 / CA4 area in the hippocampal area of SAMP8.There were discontinuous points in some neurons around the cytoplasmic nucleus. The linear colabeling site. CoIP experiment showed that CD147 interacted with mature NCT and not with immature NCT.Conclusion the expression of CD147 and NCT is higher in the pyramidal layer of hippocampus of SAMP8 mice. It is speculated that they may be closely related to the learning and memory function of hippocampus. It is suggested that CD147 may be involved in the regulation of 纬 -secretase activity.
【作者单位】：河北医科大学人体解剖学教研室;
【基金】：国家自然科学基金资助项目(31471145) 河北省自然科学基金资助项目(H2016206581) 河北省教育厅重点项目(ZD2016088) 2016年河北省研究生创新资助项目
【分类号】：R749.16

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