ApoE亚型蛋白对神经元轴突生长的影响

发布时间：2018-07-09 22:11

  本文选题：神经元 + 轴突　； 参考：《第三军医大学学报》2014年15期

【摘要】：目的观察不同亚型ApoE蛋白(apolipoprotein E,ApoE)对神经元轴突生长的影响,探讨其可能机制。方法取新生C57野生鼠和APOE基因敲除鼠大脑皮质,进行神经元原代培养;在培养第1天,在APOE基因敲除鼠神经元培养基中加入重组人类ApoE2、3、4蛋白,将所培养细胞分为ApoE(-)组、ApoE2组、ApoE3组、ApoE4组和野生组5组。倒置相差显微镜观察神经元生长情况,测量其轴突长度和突起数量。免疫荧光对神经元轴突进行染色,并对神经元轴突荧光强度进行测定;Western blot检测不同亚型ApoE对细胞分裂周期蛋白42(cell division cycle42,cdc42)表达的影响。结果培养第1、3、5天,野生组、ApoE2组、ApoE3组神经元平均轴突长度较ApoE(-)组及ApoE4组长(P0.05);培养第5天,ApoE(-)组、ApoE4组神经元突起数量分别为(1.80±0.45)个和(1.90±0.84)个,较野生组(3.80±0.84)个、ApoE2组(3.60±0.55)个、ApoE3组(3.40±1.14)个少(P0.05);野生组、ApoE2组、ApoE3组神经元轴突荧光强度分别为(54.10±7.32)、(52.40±6.33)、(50.50±8.21),较ApoE(-)组(37.20±9.30)和ApoE4组(39.00±8.32)强(P0.05);野生组、ApoE2组、ApoE3组cdc42蛋白表达高于ApoE(-)组、ApoE4组(P0.05)。而在所有实验结果中野生组、ApoE2组与ApoE3组组间以及ApoE(-)组与ApoE4组组间比较差异均无统计学意义(P0.05)。结论不同亚型ApoE蛋白对神经元轴突生长的影响不同,对tubulinⅢ及cdc42表达的影响也各不相同,后者可能是其影响神经元轴突生长的机制之一。
[Abstract]:Objective to investigate the effects of different subtypes of ApoE protein (apolipoprotein Elae) on axonal growth of neurons and its possible mechanism. Methods the cerebral cortex of newborn C57 wild mice and apo E knockout mice were harvested for primary culture, and on the first day of culture, recombinant human ApoE2O3O4 protein was added to the culture medium of apo E knockout mouse neurons. The cultured cells were divided into ApoE (-) group, ApoE _ 2 group, ApoE _ 3 group, ApoE4 group and wild group (5 groups). The neuronal growth was observed by inverted phase contrast microscope and the length of axon and the number of neurites were measured. The neuronal axons were stained with immunofluorescence and the fluorescence intensity of neuronal axons was determined by Western blot. The effect of different subtypes of ApoE on the expression of cytokinin 42 (cell division cycle42 (cdc42) was detected. Results the average axon length of ApoE3 neurons in wild ApoE2 group was significantly longer than that in ApoE (-) group and ApoE4 group (P0.05), and the number of neuronal processes in ApoE (-) group was (1.80 卤0.45) and (1.90 卤0.84), respectively. 杈冮噹鐢熺粍(3.80卤0.84)涓，

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