蛇毒神经生长因子干预HSC-T6细胞差异蛋白质分析

生物技术通报 ›› 2013, Vol. 0 ›› Issue (5): 130-136.

蛇毒神经生长因子干预HSC-T6细胞差异蛋白质分析

徐瑾¹, 张学荣¹, 胡仁统¹, 罗小玲², 陈缨¹, 林兴¹, 廖明¹, 付娆¹, 付道滢¹

(1.广西医科大学医学科学实验中心，南宁 530021；2.广西医科大学肿瘤医院，南宁 530021)

收稿日期:2012-11-20 修回日期:2013-05-24 出版日期:2013-05-24 发布日期:2013-05-24
作者简介:徐瑾，女，硕士研究生，研究方向：眼镜蛇毒NGF与肝纤维化蛋白质组学；E-mail：576776818@qq.com
基金资助:
国家自然科学基金资助项目(81160063)，广西自然科学基金资助项目(2011GXNSFA018268，KFJJ2010-43)，广西教育厅科研项目(201012MS058)，广西中医药管理局项目(GZKZ 10-013)

Analysis for Differentially Expressed Proteins of HSC-T6 Cell Affected by Nerve Growth Factor

Xu Jin¹, Zhang Xuerong¹, Hu Rentong¹, Luo Xiaoling², Chen Ying¹, Lin Xing¹, Liao Ming¹, Fu Rao¹, Fu Daoying¹

(1. Medical Sciences Experimental Center of Guangxi Medical University，Nanning 530021；2. Cancer Hospital of Guangxi Medical University，Nanning 530021)

Received:2012-11-20 Revised:2013-05-24 Published:2013-05-24 Online:2013-05-24
About author:张学荣，男，教授，硕士生导师，研究方向：蛇毒药物应用，肝纤维化；E-mail：zxrsv@sina.com

摘要/Abstract

摘要： 旨在建立肝星状细胞(HSC-T6)双向凝胶电泳图谱，初步分析NGF对HSC-T6细胞蛋白质表达的影响。试验设立空白对照组和NGF(4、8和16 mg/L)处理组，分别作用于HSC-T6细胞，24 h后提取细胞总蛋白；利用2-DE分离空白对照组和NGF处理组细胞总蛋白质后，经图像分析识别差异表达的蛋白点，应用基质辅助激光解吸电离飞行时间质谱鉴定差异蛋白质；生物信息学对差异蛋白质点进行GO分类及信号传导通路分析。结果显示，比较分析空白对照组和NGF作用组的2-DE图谱，找到差异蛋白质点47个，其中在NGF作用组表达上调22个，下调25个，差异蛋白质点的表达部分随着NGF浓度的增加呈递增性，部分随着NGF浓度的增加呈递减性，对其中18个表达差异1.8倍以上的蛋白质点进行肽质量指纹图分析，鉴定出13个与细胞信号传导、细胞增殖、氧化代谢有关的蛋白质。眼镜蛇毒NGF能改变HSC-T6细胞信号通路中相关蛋白质的差异表达，为在蛋白质水平研究NGF抗肝纤维化机制提供理论依据。

关键词: 眼镜蛇毒(NGF), 肝星状细胞(HSC-T6), 蛋白质组学, 双向电泳技术, 飞行时间串联质谱

Abstract: It was to develop the 2-DE profiles of proteome from hepatic stellate cell and preliminarily analyze the affect of NGF on the protein expression of HSC-T6 cell. We established groups of control and NGF (4、8、16 mg/L), respectively, then acting on the HSC-T6. The differential expression of proteins were analyzed by imaging analysis and MALDI-TOF-MS after the total protein was extracted from the blank control group and NGF treated by 2-DE. The differential protein spots were classified with GO and analyzed with Signal transduction pathway. Results showed that HSC-T6 cell proliferation is inhibited evidently. Forty-seven differentially expressed protein were found in the proteome profile analysis of these two types of blank control group and NGF treated, among which 22 protein spots were up-regulated and 25 protein spots were down-regulated in the HSC-T6 treated by NGF, part of the different expression of protein is increasing with increasing of NGF concentration and part is decreasing with increasing of NGF concentration. Eighteen protein spots with the differential expressions more than 1.8 times were analyzed by peptide mass fingerprint (PMF) and thirteen differentially expressed proteins were found to be related with cell signal transduction, cell proliferation and oxidative stress. The cobra venom NGF can change the expression of the different protein expression of HSC-T6 cell in signaling pathway, providing a theoretical basis for the antifibrotic mechanism of NGF at protein level.

Key words: NGF, HSC-T6, Proteomics 2-DE, MALDI-TOF-MS

徐瑾, 张学荣, 胡仁统, 罗小玲, 陈缨, 林兴, 廖明, 付娆, 付道滢. 蛇毒神经生长因子干预HSC-T6细胞差异蛋白质分析[J]. 生物技术通报, 2013, 0(5): 130-136.

Xu Jin, Zhang Xuerong, Hu Rentong, Luo Xiaoling, Chen Ying, Lin Xing, Liao Ming, Fu Rao, Fu Daoying. Analysis for Differentially Expressed Proteins of HSC-T6 Cell Affected by Nerve Growth Factor[J]. Biotechnology Bulletin, 2013, 0(5): 130-136.

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