转录因子X盒结合蛋白1转染对神经干细胞在缺氧环境下grp78、EDEM表达及抗凋亡能力的影响  

Effects of transfected adenovirus-mediated transcription factor X-box binding protein 1 on expression of grp78,EDEM and hippocampal-derived neural stem cell apoptosis under hypoxia

作　　者：金鲜花[1] 郑胜哲[1] 卢蕾[2] 宋磊[2] 

机构地区：[1]延边大学附属医院神经内科,吉林延边133000 [2]吉林大学第一医院神经内科,吉林长春130021

出　　处：《中风与神经疾病杂志》2010年第9期791-795,共5页Journal of Apoplexy and Nervous Diseases

摘　　要：目的将X盒结合蛋白1基因以重组腺病毒为载体转染胚鼠海马神经干细胞,观察其在缺氧环境下是否可以使移植后干细胞内grp78、EDEM表达增加以及对干细胞抗凋亡能力的影响。方法取孕16d SD大鼠胚鼠的海马组织进行神经干细胞的分离、克隆、nestin免疫荧光检测,以及传代和扩增。将重组腺病毒Ad-XBP1-EGFP质粒转染胚鼠海马神经干细胞,得到基因修饰后的胚鼠海马神经干细胞。选取未转染的神经干细胞标记为对照组、未转染组;选取转染后的神经干细胞标记为转染组。未转染组和转染组用CoCl2诱导缺氧,24h后WesternBlot检测3组神经干细胞中xbp-1,grp78,EDEM,bcl-2及bax表达,流式细胞术检测NSCs和XBP1-NSCs的凋亡情况。结果在缺氧条件下,与未转染组相比,转染组的grp78表达增加,EDEM表达水平上升,bcl-2水平上升,而bax水平下降(P<0.05);转染组神经干细胞凋亡程度较未转染组减轻(P<0.05)。结论可以利用重组腺病毒作为载体成功将X盒结合蛋白1基因导入胚鼠海马神经干细胞中;转染后的神经干细胞在缺血缺氧条件下grp78、EDEM水平上升,抗凋亡能力较普通神经干细胞明显增加。Objective To determine the effects of the XBP1 gene on NSC apoptosis under hypoxic conditions following XBP1 gene transfection into rat embryonic hippocampal NSCs using recombinant adenovirus vector.Methods Hippocampi from embryonic,Sprague Dawley rats on gestational 16d were harvested for NSC isolation and cloning,followed by immunofluorescence for Nestin and sub-culturing.The recombinant adenovirus Ad-XBP1-enhanced green fluorescent protein plasmid was transfected into rat embryonic hippocampal NSCs,and then CoCl2 was applied to induce hypoxia.Western blot assay was utilized to quantify XBP1,GRP78,EDEM,Bcl-2,and Bax expression.Flow cytometry was used to measure apoptosis.Results Under hypoxic conditions,Grp78,EDEM,and Bcl-2 levels increased,but Bax levels decreased.In addition,NSC apoptosis decreased following transfection（P〈0.05）.Conclusion The XBP1 gene was successfully transfected into rat embryonic hippocampal NSCs using a recombinant adenovirus vector.NSC anti-apoptotic effects under hypoxia were significantly increased following transfection.

关 键 词：X盒结合蛋白1 凋亡 内质网应激 神经干细胞 移植 

分 类 号：Q813[生物学—生物工程]