摘要
目的鉴定分析神经再生条件液(Nerver egeneration conditioned fluid,NRCF)中相对分子质量为(232～440)×103区域的蛋白条带,并探索NRCF中补体C3可能的促神经再生作用。方法采用Shotgun蛋白质组学策略、液相色谱一串联质谱对相对分子质量为(232～440)×103区域的蛋白条带的组分进行定性分析。在原代培养的皮层神经元中分别加入不同浓度的外源性补体C3蛋白和神经生长因子,以Neurolucida神经元分析软件采集各实验组每个大脑皮层神经元最长突起长度。结果NRCF中相对分子质量在(232～440)×103的蛋白条带共鉴定到54种蛋白质,其中包括补体C3α链,补体C3α链和神经突起导向因子Netrins结构中都含有一个共同功能域NTR组件。培养液加入特定浓度补体C3(0.5～5μg/mL)后培养96h的神经元突起长度较正常对照组显著增长(P<0.05)。结论补体C3存在于NRCF中,特定浓度补体C3对神经元突起有明显促进生长作用。
Objective To explore the effects of promoting nerve regeneration of complement C3 in nerve regeneration conditioned fluid （NRCF）. Methods The protein band of the relative molecular mass of （232-440）×10^3 in NRCF were analyzed by the Shotgun technique, liquid chromatography and mass spectrometry （LC-MS/MS）. The primary cultured cortical neurons were exposed to different concentrations of exogenous complement C3 and nerve growth factor （NGF）. Photograph of neurons in each group was taken every day, and the length of the longest neurite of each neuron was calculated by a neuron morphology analysis software. The neurons without exposure to the other agents and the culture fluid were served as controls. Results 54 proteins were identified in the protein band of the relative molecular mass of （232-440）×10^3. Complement C3 alphachain protein, which had a NTR module as well as neutrins in structure, was included in these proteins. The neurites of neurons with the presence of low C3 concentrations were significant longer than the neurites of normal control group after culturing 96 hours. Conclusion There was complement C3 protein in NRCF. Low concentration of complement C3 has positive effect on neurites growth in cultured rat cortical neurons. It implied that the complement C3 has a positive effect of promoting nerve regeneration in NRCF.
出处
《组织工程与重建外科杂志》
2009年第1期16-20,共5页
Journal of Tissue Engineering and Reconstructive Surgery
基金
国家自然科学基金(30471795)
关键词
神经再生条件液
神经再生
补体C3
神经突
Nerve regeneration conditioned fluid （NRCF）
Nerve regeneration
Complement C3
Neurite
