SELDI蛋白芯片技术检测叶酸对胎鼠神经干细胞作用的蛋白表达谱  

DETECTION OF THE PROTEINS PROFILE OF FETAL NEURAL STEM CELLS TREATED WITH FOLIC ACID BY SELDI PROTEIN CHIP TECHNOLOGY IN RAT

作　　者：张莹[1] 常红[1] 黄国伟[1] 张绪梅[1] 

机构地区：[1]天津医科大学公共卫生学院营养与食品卫生教研室,天津300070

出　　处：《营养学报》2011年第2期188-192,195,共6页Acta Nutrimenta Sinica

基　　金：国家自然科学基金(No.30901192;30771797)

摘　　要：目的探讨应用表面增强激光解离飞行时间质谱技术(surface-enhanced laser desorption/ionization time of flight mass spectrometry,SELDI-TOF-MS)测试不同剂量叶酸作用下胎鼠神经干细胞(NSCs)的蛋白质表达谱,从中筛选出差异表达蛋白。方法原代培养胎鼠神经干细胞,分为4组:对照组(叶酸浓度为4μg/ml),叶酸缺乏组(叶酸浓度为0.65μg/ml),叶酸低剂量组(叶酸浓度为8μg/ml),叶酸高剂量组(叶酸浓度为44μg/ml);细胞形态学观察;MTT法检测叶酸对细胞增殖状态的影响;采用弱阳离子交换芯片(wcx-2)蛋白芯片检测不同叶酸剂量下胎鼠NSCs蛋白表达差异;采用Biomarker Wizard软件进行分析结果,通过查询蛋白库,对特定分子质量所对应的标记蛋白进行初步确定。结果叶酸补充可促进NSCs增殖;每组芯片上均捕获到102种蛋白质点。筛选出不同叶酸剂量组(叶酸缺乏组、叶酸低剂量组、叶酸高剂量组)与对照组之间的差异蛋白共32种,其中有4个蛋白存在显著性差异(P<0.05)。初步推断这些蛋白与NSCs凋亡相关,且在不同叶酸剂量组图谱表达不同。结论叶酸可能通过改变凋亡蛋白的表达影响NSCs生长。叶酸对神经系统疾病的防治作用将进一步探索。Objective To detect the fetal rat neural stem cell（NSC） protein profiling treated with different doses of folic acid（FA） by surface-enhanced laser desorption/ionization time of flight mass spectrometry（SELDI-TOF-MS） and find out the differential proteins.Method Primary cultured fetal rat NSC were divided into 4 groups： control group（FA 4μg/ml in media）,FA-deficient group（FA 0.65μg/ml in media）,low FA group（FA 8μg/ml in media）,high FA group（FA 44μg/ml in media）.NSC proliferation was detected after treatment of FA by methylthiazoltatragolium（MTT） technique.Wcx-2 chip was used for discovering the discriminatory proteins,and the results were analyzed by Biomarker Wizard software.Differential proteins were initially identified through querying the protein database.Results MTT assay showed that the growth tendency was increased with folate concentration in the media.102 proteins were differentially expressed on the chips.Compared with the control group,32 proteins were differentially expressed in other groups,and 4 proteins of them had significant differences（P〈0.05）.These proteins were primarily considered to be associated with apoptosis in NSCs,and expressed differently in protein atlas of different FA groups.Conclusion Folic acid may alter the expression of apoptic protein during the NSCs growth.The effect of FA in prevention and treatment of nervous system diseases will be further explored.

关 键 词：叶酸 神经干细胞 蛋白芯片 胎鼠 

分 类 号：R151[医药卫生—营养与食品卫生学]