低浓度补体C3对大脑皮层神经元生物活性及突起生长的影响  

Effects of complement C_3 exposure on survival and neurite growth in primary rat cortical neuron cultures

作　　者：李玉萍[1] 孟繁君[1] 陆佩华[2] 李青峰[1] 

机构地区：[1]上海交通大学医学院第九人民医院整复外科,上海200011 [2]上海交通大学基础医学院神经生物学实验室,上海200025

出　　处：《上海交通大学学报（医学版）》2008年第8期953-956,共4页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家自然科学基金(30471795)~~

摘　　要：目的探讨补体C3对大脑皮层神经元生物活性和突起生长的影响及可能机制。方法在原代培养的孕16 d大鼠胚胎皮层神经元中分别加入不同浓度的外源性C3蛋白、神经生长因子和C3a受体拮抗剂SB290157。WST-8法测定神经元活性（细胞存活率）。选用MAP2与β-tubulin免疫荧光细胞化学双标染色,荧光显微镜下观察并计算神经元纯度;应用神经元形态分析软件采集神经元树突、轴突长度及胞体面积等数据,分析不同浓度补体C3及C3a受体拮抗剂对突起生长的影响。以不含其他试剂的神经元细胞及其培养液作为正常对照组。结果皮层神经元存活率检测显示,高浓度（10-100μg/mL）C3组明显低于正常对照组（P〈0.05）,而低浓度（0.01-1μg/mL）C3组与对照组无明显差异。1μg/mL和5μg/mL C3均促进皮层神经元突起生长,尤其是轴突的生长,其树突和轴突长度分别增长20%-39%和40%-61%,与正常对照组比较有显著差异（P〈0.05和P〈0.01）;而加入C3a受体拮抗剂后,皮层神经元树突和轴突长度与对照组比较无明显差异。结论低浓度补体C3对皮层神经元轴突和树突生长有明显促进作用,而该作用可能由C3活性片段C3a受体介导。Objective To investigate the effects of complement C3 exposure on viability and neurite growth in primary rat cortical neuron cultures,and to explore the possible mechanisms. Methods The cortical neurons of embryonic rats with pregnancy for 16 d were exposed to different concentrations of exogenous C3 protein,nerve growth factor and C3α receptor antagonist SB290157.Cell viability was assessed by WST-8 assay.MAP2 and β-tubulin immunofluorescence cytochemical staining were employed to observe and calculate the neuron purity.A neuron morphology analysis software was adopted to collect the data such as neuron dendrite length,axonal length and cell body area for the analysis of effects of different concentrations of complement C3 and C3a receptor antagonist on the neurite growth.The neurons without exposure to the other agents and the culture fluid were served as controls. Results The cortical neuron viability was significantly lower in groups with higher concentrations of C3（10-100 μg/mL） than that in control group（P〈0.05）,while there was no significant difference between groups with lower concentrations of C3（0.01-1 μg/mL） and control group.Significant effects on neurite length were observed in the presence of C3 of 1 μg/mL and 5 μg/mL,where C3-induced elongation of axons（40%-61%） and dendrites（20%-39%） became more apparent（P〈0.01 and P〈0.05 vs control group）.This potentiation of C3-induced neurite elongation was significantly inhibited by C3a receptor antagonist SB290157（P〉0.05 vs control group）. Conclusion Lower concentrations of C3 have positive effects on neurite growth in cultured rat cortical neurons,and the effects may be mediated by the C3a receptor. ：

关 键 词：神经元 补体C3 神经突起 C3a受体 

分 类 号：Q42[生物学—神经生物学]