摘要
目的:研究维甲酸(RA)诱导PC12细胞向神经元样细胞分化过程中的细胞直径和突起长度的变化情况以及诱导分化的PC12细胞MAP2的表达情况。方法:实验共分7组,分别为0.1、0.3、0.5、1.0、2.0及5.0mg.L-1RA组及含10%胎牛血清的对照组。不同浓度的RA组均诱导PC12细胞72h,在24、48和72h分别观察细胞的突起长度和细胞最大直径的变化情况;在诱导72h后利用免疫细胞化学技术,观察不同剂量RA作用后PC12细胞向MAP2阳性细胞分化的情况。结果:0.3、0.5、1.0、2.0mg.L-1RA组MAP2阳性细胞数与对照组比较明显增加,且以1.0mg.L-1组最为明显(P<0.05)。与对照组比较,加入RA组分化的细胞状态较好,且突起和细胞直径明显增长和增大,以1.0mg.L-1组最为明显(P<0.01)。但是随着诱导时间的延长,2.0mg.L-1浓度的RA即可导致细胞中黑色颗粒增多,胞体回缩,细胞老化,视野中多为细胞碎屑颗粒,有的细胞融合成片。结论:RA具有诱导PC12细胞向神经元样细胞分化的趋势,且能促进分化细胞突起的生长和细胞直径的增大。1.0mg.L-1RA浓度是诱导PC12细胞向神经元样细胞分化的最适浓度。
Objective To study the condition of pheochromocytoma cells （PC12 cells） differentiated into neuron-like cells induced by retinoic acid （RA） and to observe the lengh of neurite and max diameter and the expression of MAP2 in PC12 cells during their differentiation into neuron-like cells. Methods There were seven groups ： 0. 1, 0.3, 0.5, 1.0, 2.0 and 5.0 mg·L^-1RA groups and control group which contained 10% fetal calf serum. PC12 cells were exposed to RA with different concentrations for 72 h and the lengh of neurite and max diameter of PC12 cells were observed at 24, 48, and 72 h respectively. The expression of MAP2 was detected by immunocytochemistry after PC12 cells were exposed to RA for 72 h. Results The number of MAP2 positive cells was significantly increased in 0. 3, 0.5, 1.0, 2.0 mg·L^-1 RA groups compared with control group and the most significant concentration was 1.0 mg·L^-1 （P〈0. 05） . The length of total primary neurite and max diameter of PC12 cells in 0.3, 0.5, 1.0, 2.0 mg·L^-1RA groups were obviously longer and bigger than those in control group and the most significant concentration also was 1.0 mg·L^-1 （P〈0.01）. But the number and the lengh of neurite were fewer and shorter when PC12 cells were exposed to 2.0 mg·L^-1 RA than that of 1.0 mg·L^-1 RA. There were a lot of black particles in PC12 cells and cells became smaller and aging when thev were exposed to 2.0 mg ·L^-1RA for 7 d. Conclusion PC12 cells could differentiate into neuron-like in vitro induced by RA, and RA could enhance the outgrowth of neurite and multiplication of max diameter of the differentiated PC12 cells. 1.0 mg ·L^-1 is the optimal concentration in which RA could induce PC12 cells to differentiate into neuron-like cells.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2008年第2期187-190,F0002,共5页
Journal of Jilin University：Medicine Edition
基金
教育部高等学校博士学科点专项科研基金资助课题(20030183048)
吉林大学种子基金资助课题(2005)
