中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2013年
37期
6613-6619
,共7页
陈衍晨%赵丹%卿娣%程东良%毛姣玉%王斌
陳衍晨%趙丹%卿娣%程東良%毛姣玉%王斌
진연신%조단%경제%정동량%모교옥%왕빈
组织构建%组织构建细胞学实验%c-fos基因%神经细胞凋亡%缺氧再灌注%脑损伤%斑马鱼%胚胎%新生儿%分子生物学
組織構建%組織構建細胞學實驗%c-fos基因%神經細胞凋亡%缺氧再灌註%腦損傷%斑馬魚%胚胎%新生兒%分子生物學
조직구건%조직구건세포학실험%c-fos기인%신경세포조망%결양재관주%뇌손상%반마어%배태%신생인%분자생물학
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。<br> 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。<br> 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。<br> 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。
揹景:國外已經有學者使用斑馬魚胚胎開始進行缺氧再灌註的研究,但還沒有關于c-fos基因在斑馬魚腦缺氧再灌註過程中的錶達及其作用機製的報道。<br> 目的:觀察缺氧再灌註後斑馬魚胚胎腦部細胞凋亡及腦組織中c-fos基因的錶達情況。<br> 方法:取48 hpf的斑馬魚胚胎進行缺氧實驗,模擬新生兒缺氧再灌註損傷環境,通過嚮水中通入99.999%高純氮氣製造缺氧環境,分彆經過6,12,24 h的缺氧處理後,在正常氧體積分數下進行6 h恢複。對照組為正常通氣組(溶解氧濃度在7.0 mg/L左右)。採用吖啶橙染色方法,觀察不同缺氧時間對斑馬魚神經細胞凋亡的影響,同時採用實時熒光定量覈痠擴增檢測繫統(qPCR),對 c-fos 基因錶達情況進行定量分析,比較缺氧再灌註前後c-fos基因錶達水平的變化。<br> 結果與結論:對照組腦部能檢測到微量細胞凋亡,c-fos基因呈低水平錶達;實驗組經過6,12,24 h缺氧後,腦部凋亡細胞逐漸增多,缺氧24 h組凋亡細胞增幅最大(P<0.05),c-fos基因錶達有不同程度升高(P<0.05),尤其是缺氧6 h後,該基因的錶達上調幅度最高。結果錶明缺氧會導緻斑馬魚腦細胞內c-fos基因錶達上調,可能是導緻缺氧後期腦細胞凋亡激增的機製之一。
배경:국외이경유학자사용반마어배태개시진행결양재관주적연구,단환몰유관우c-fos기인재반마어뇌결양재관주과정중적표체급기작용궤제적보도。<br> 목적:관찰결양재관주후반마어배태뇌부세포조망급뇌조직중c-fos기인적표체정황。<br> 방법:취48 hpf적반마어배태진행결양실험,모의신생인결양재관주손상배경,통과향수중통입99.999%고순담기제조결양배경,분별경과6,12,24 h적결양처리후,재정상양체적분수하진행6 h회복。대조조위정상통기조(용해양농도재7.0 mg/L좌우)。채용아정등염색방법,관찰불동결양시간대반마어신경세포조망적영향,동시채용실시형광정량핵산확증검측계통(qPCR),대 c-fos 기인표체정황진행정량분석,비교결양재관주전후c-fos기인표체수평적변화。<br> 결과여결론:대조조뇌부능검측도미량세포조망,c-fos기인정저수평표체;실험조경과6,12,24 h결양후,뇌부조망세포축점증다,결양24 h조조망세포증폭최대(P<0.05),c-fos기인표체유불동정도승고(P<0.05),우기시결양6 h후,해기인적표체상조폭도최고。결과표명결양회도치반마어뇌세포내c-fos기인표체상조,가능시도치결양후기뇌세포조망격증적궤제지일。
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. <br> OBJECTIVE:To observe the zebrafish embryonic brain cel apoptosis and expression of c-fos gene in brain tissues after hypoxia reperfusion. <br> METHODS:Zebrafish embryos were selected at 48 hours post fertilization. Neonatal hypoxia reperfusion injury was simulated by gradual y 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 amplification detection system. And the expression level of c-fos gene was compared before and after hypoxia reperfusion. <br> RESULTS AND CONCLUSION:A smal amount of apoptotic brain cel s could be detected in the control group, and the c-fos gene expression level was decreased;in the experimental group, the number of apoptotic cel s was increased after hypoxia for 6, 12 and 24 hours, and the gene expression after hypoxia for 6 hours was increased distinctly. The results indicate that hypoxia can increase the c-fos gene expression in brain cel s of zebrafish embryos, which may be one of the mechanisms of brain cel apoptosis increasing after hypoxia.