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。
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.
出处
《生物技术通报》
CAS
CSCD
北大核心
2013年第5期130-136,共7页
Biotechnology Bulletin
基金
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)
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
通讯作者：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
