發(fā)布時(shí)間：2018-07-09 22:11

  本文選題：神經(jīng)元 + 軸突��； 參考：《第三軍醫(yī)大學(xué)學(xué)報(bào)》2014年15期

【摘要】：目的觀察不同亞型ApoE蛋白(apolipoprotein E,ApoE)對神經(jīng)元軸突生長的影響,探討其可能機(jī)制。方法取新生C57野生鼠和APOE基因敲除鼠大腦皮質(zhì),進(jìn)行神經(jīng)元原代培養(yǎng);在培養(yǎng)第1天,在APOE基因敲除鼠神經(jīng)元培養(yǎng)基中加入重組人類ApoE2、3、4蛋白,將所培養(yǎng)細(xì)胞分為ApoE(-)組、ApoE2組、ApoE3組、ApoE4組和野生組5組。倒置相差顯微鏡觀察神經(jīng)元生長情況,測量其軸突長度和突起數(shù)量。免疫熒光對神經(jīng)元軸突進(jìn)行染色,并對神經(jīng)元軸突熒光強(qiáng)度進(jìn)行測定;Western blot檢測不同亞型ApoE對細(xì)胞分裂周期蛋白42(cell division cycle42,cdc42)表達(dá)的影響。結(jié)果培養(yǎng)第1、3、5天,野生組、ApoE2組、ApoE3組神經(jīng)元平均軸突長度較ApoE(-)組及ApoE4組長(P0.05);培養(yǎng)第5天,ApoE(-)組、ApoE4組神經(jīng)元突起數(shù)量分別為(1.80±0.45)個和(1.90±0.84)個,較野生組(3.80±0.84)個、ApoE2組(3.60±0.55)個、ApoE3組(3.40±1.14)個少(P0.05);野生組、ApoE2組、ApoE3組神經(jīng)元軸突熒光強(qiáng)度分別為(54.10±7.32)、(52.40±6.33)、(50.50±8.21),較ApoE(-)組(37.20±9.30)和ApoE4組(39.00±8.32)強(qiáng)(P0.05);野生組、ApoE2組、ApoE3組cdc42蛋白表達(dá)高于ApoE(-)組、ApoE4組(P0.05)。而在所有實(shí)驗(yàn)結(jié)果中野生組、ApoE2組與ApoE3組組間以及ApoE(-)組與ApoE4組組間比較差異均無統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論不同亞型ApoE蛋白對神經(jīng)元軸突生長的影響不同,對tubulinⅢ及cdc42表達(dá)的影響也各不相同,后者可能是其影響神經(jīng)元軸突生長的機(jī)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 鹵0.45) and (1.90 鹵0.84), respectively. 杈冮噹鐢熺粍(3.80鹵0.84)涓，

本文編號：2110856