斑马鱼体内氧化损伤标志物筛选  

Screening for markers of oxidative damage in zebrafish

作　　者：李嘉蔚 胡雪菲 韩鹏 陈呢喃 陆一鸣[1,2,3] LI Jiawei;HU Xuefei;HAN Peng;CHEN Ninan;LU Yiming(Sino-French Research Center for Life Sciences and Genomics,Ruijin Hospital,School of Medicine,Shang-hai Jiaotong University,Shanghai 200025;Department of Emergency,Ruijin Hospital North Branch,School of Medicine,Shanghai Jiaotong University,Shanghai 200025;Department of Emergency,Ruijin Hospital,School of Medicine,Shanghai Jiaotong University,Shanghai 200025,China)

机构地区：[1]上上海交通大学医学院附属瑞金医院中法生命科学和基因组研究中心,海200025 [2]上海交通大学医学院附属瑞金医院北院急诊科,上海200025 [3]上海交通大学医学院附属瑞金医院急诊科,上海200025

出　　处：《郑州大学学报（医学版）》2018年第4期499-502,共4页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金项目(81171846,81671900)

摘　　要：目的:探讨以端粒为靶点的氧化损伤标志物的筛选,进而可能可以用来临床上早期发现或预报氧化损伤。方法:采用端粒重复序列结合因子2(TRF2)基因敲除(TRF2-/-)斑马鱼胚胎作为氧化损伤模型,RT-q PCR法检测野生型和TRF2-/-斑马鱼胚胎中TRF2、PUM3、PACSIN3 mRNA的表达情况。构建pCS2-PUM3、pCS2-PACSIN3质粒并体外转录后,将mRNA分别显微注射至TRF2-/-斑马鱼受精卵中,比较注射组与未注射组斑马鱼畸形率。结果:RT-q PCR结果表明,与野生型斑马鱼胚胎相比,TRF2-/-斑马鱼胚胎TRF2、PUM3、PACSIN3 mRNA的表达量下降(P<0.05)。显微注射PUM3 mRNA至TRF2-/-斑马鱼受精卵后斑马鱼胚胎畸形率下降(P<0.05)。结论:TRF2能影响氧化应激相关基因PUM3、PACSIN3的表达,且PUM3能补救TRF2缺失造成的氧化损伤相关器官畸形。Aim： To screen oxidative damage markers targeting telomeres,which may then be used to clinically detect or predict oxidative damage early. Methods： The telomeric repeat binding factor 2（ TRF2） knockout（ TRF2-/-） zebrafish embryos were used as an oxidative damage model. RT-q PCR was used to detect the expression of TRF2,PUM3 and PACSIN3 mRNA in wild-type and TRF2-/-zebrafish embryos. After p CS2-PUM3 and p CS2-PACSIN3 plasmids were constructed and in vitro transcribed,their mRNA were into TRF2-/-zebrafish fertilized eggs,and the zebrafish malformation rates in the groups with injection and without injection were summarized and statistically analyzed. Results： RT-q PCR results showed that the TRF2,PUM3,and PACSIN3 mRNA expression levels in TRF2-/-zebrafish embryos were significantly lower than those in wild-type zebrafish embryos（ P 0. 05）. After microinjection of zebrafish PUM3 mRNA into TRF2-/-zebrafish fertilized eggs,the malformation rates of zebrafish embryos decreased（ P 0. 05）. Conclusion： TRF2 can affect the expressions of oxidative stress related genes PUM3 and PACSIN3,and PUM3 can remedy the oxidative damage-related organ malformation caused by TRF2 deficiency. Targeting these molecules is expected to detect or predict oxidative stress early in clinical field.

关 键 词：斑马鱼 氧化应激 端粒 端粒重复序列结合因子2 

分 类 号：R33[医药卫生—人体生理学]