缺氧再灌注斑马鱼胚胎脑部细胞凋亡及c-fos基因的表达  

Zebrafish embryonic brain cell apoptosis and c-fos gene expression after hypoxia reperfusion

作　　者：陈衍晨[1] 赵丹[1] 卿娣[1] 程东良[1] 毛姣玉[1] 王斌[1] 

机构地区：[1]南方医科大学珠江医院儿科中心,广东省广州市510282

出　　处：《中国组织工程研究》2013年第37期6613-6619,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:国外已经有学者使用斑马鱼胚胎开始进行缺氧再灌注的研究,但还没有关于c-fos基因在斑马鱼脑缺氧再灌注过程中的表达及其作用机制的报道。目的:观察缺氧再灌注后斑马鱼胚胎脑部细胞凋亡及脑组织中c-fos基因的表达情况。方法:取48 hpf的斑马鱼胚胎进行缺氧实验,模拟新生儿缺氧再灌注损伤环境,通过向水中通入99.999%高纯氮气制造缺氧环境,分别经过6,12,24 h的缺氧处理后,在正常氧体积分数下进行6 h恢复。对照组为正常通气组(溶解氧浓度在7.0 mg/L左右)。采用吖啶橙染色方法,观察不同缺氧时间对斑马鱼神经细胞凋亡的影响,同时采用实时荧光定量核酸扩增检测系统(qPCR),对c-fos基因表达情况进行定量分析,比较缺氧再灌注前后c-fos基因表达水平的变化。结果与结论:对照组脑部能检测到微量细胞凋亡,c-fos基因呈低水平表达;实验组经过6,12,24 h缺氧后,脑部凋亡细胞逐渐增多,缺氧24 h组凋亡细胞增幅最大(P<0.05),c-fos基因表达有不同程度升高(P<0.05),尤其是缺氧6 h后,该基因的表达上调幅度最高。结果表明缺氧会导致斑马鱼脑细胞内c-fos基因表达上调,可能是导致缺氧后期脑细胞凋亡激增的机制之一。BACKGROUND： Foreign scholars have researched hypoxia reperfusion in zebrafish embryos, but there is no research on c-fos gene expression and the mechanism during zebrafish cerebral hypoxia reperfusion. OBJECTIVE： To observe the zebrafish embryonic brain cell apoptosis and expression of c-fos gene in brain tissues after hypoxia reperfusion. ME＇IFHOOS： Zebrafish embryos were selected at 48 hours post fertilization. Neonatal hypoxia reperfusion injury was simulated by gradually leading nitrogen （99.999%） into the device. After hypoxia treatment for 6, 12 and 24 hours, the embryos received reperfusion for 6 hours under normal oxygen concentration. The embryos in the control group received normoventilation （the dissolved oxygen concentration was about 7.0 mg/L）. Acridine orange staining was performed to observe the effect of different hypoxia durations on the apoptosis of neurons in zebrafish, and then the c-fos gene expression was quantitative analyzed with real-time quantitative nucleic acid amplificat^n detection system. And the expression level of c-fos gene was compared before and after hypoxia reperfusion. RESULTS AND CONCLUSION： A small amount of apoptotic brain cells could be detected in the control group, and the c-fos gene expression level was decreased; in the experimental group, the number of apoptotic cells was increased after hypoxia for 6, 12 and 24 hours, and the gene expression after hypoxia for 6 hours was Incraased distinctly. The results indicate that hypoxla can increase the c.fos gene expression irt brain cells of zebraflsh embryos, which may be one of the mechanisms of brain cell apoptosis increasing after hypoxia.

关 键 词：基因 fos 细胞凋亡 缺氧 再灌注 斑马鱼 

分 类 号：R318[医药卫生—生物医学工程]