宋锦;李莉;尚鲁俊;吴西军;周艳华;何志旭;舒莉萍;
SONG Jin;LI Li;SHANG Lujun;WU Xijun;ZHOU Yanhua;HE Zhixu;SHU Liping;Department of Immunology,Guizhou Medical University;The Center of Stem Cell and Tissue Engineering Research,Guizhou Medical University;Department of Paediatrics,Guizhou Medical University;Experimental Animal Center,Guizhou Medical University,Guizhou Medical University;

• 1:贵州医科大学免疫学教研室
• 2:贵州医科大学干细胞与组织工程实验中心
• 3:贵州医科大学
• 4:贵州医科大学儿科学教研室

摘要(Abstract)：

目的:观察不同发育时期的斑马鱼胚胎、成年斑马鱼不同组织的G6PD酶活性及g6pd mRNA的表达。方法:收集受精后24 h(24 hpf)、48 hpf、受精后5 d(5 dpf)及15 dpf的野生型斑马鱼胚胎,同时收集成年斑马鱼的脑、肌肉、鳃、皮肤、眼睛及肠等组织,运用改良G6PD定量比值法和RT-PCR法检测上述胚胎或组织的G6PD酶活性及g6pd mRNA表达水平。结果:G6PD酶活性、g6pd mRNA表达水平在24 hpf、48 hpf、5 dpf及15 dpf比较,差异无统计学意义(P>0.05);在成年斑马鱼的脑、肌肉、鳃、皮肤、眼睛及肠等不同部位组织比较,差异也无统计学意义(P>0.05)。结论:野生斑马鱼G6PD酶活性、g6pd mRNA表达水平不受胚胎发育时期和组织部位的影响。
Objective: To observe G6 PD enzyme activity and G6 PD mRNA expression in different tissues of zebrafish embryos and adult zebrafish at different developmental stages. Method: After fertilization,24 hpf( 24 hour post-fertilization),48 hpf,5 dpf( 5 day post-fertilization) and 15 dpf wild type zebrafish embryos were collected,and the brain,muscle,gill,skin,eyes and intestine of adult zebrafish were collected simultaneously. The modified G6 PD quantitative ratio assay and RT-PCR were adopted to detect G6 PD activity and mRNA expression levels in zebrafish embryos and tissues. Result:There was no statistically significant difference in G6 PD activity and mRNA expression levels between24 hpf,48 hpf,5 dpf and 15 dpf( P > 0. 05). There was also no statistically significant difference in G6 PD activity and mRNA expression levels between brain,muscle,gill,skin,eyes and intestine( P >0. 05). Conclusion: The G6 PD activity and mRNA expression level of wild zebrafish is not affected by embryonic development period and tissue site.

关键词(KeyWords)： 斑马鱼;胚胎;葡萄糖6磷酸脱氢酶;酶活性;mRNA
zebrafish;embryo;glucose-6-phosphate dehydrogenase;enzymatic activity;mRNA

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目资助(31360285)

作者(Author): 宋锦;李莉;尚鲁俊;吴西军;周艳华;何志旭;舒莉萍;
SONG Jin;LI Li;SHANG Lujun;WU Xijun;ZHOU Yanhua;HE Zhixu;SHU Liping;Department of Immunology,Guizhou Medical University;The Center of Stem Cell and Tissue Engineering Research,Guizhou Medical University;Department of Paediatrics,Guizhou Medical University;Experimental Animal Center,Guizhou Medical University,Guizhou Medical University;

Email:

DOI: 10.19367/j.cnki.1000-2707.2016.07.002

参考文献(References)：

• [1]Santana MS,Monteiro WM,Siqueira AM,et al.Glucose-6-phosphate dehydrogenase deficient variants are associated with reduced susceptibility to malaria in the Brazilian Amazon[J].Trans RSoc Trop Med Hyg,2013(5):301-306.
• [2]Howes RE,Piel FB,Patil AP,et al.G6PD deficiency prevalence and estimates of affected populations in malaria endemic countries:a geostatistical model-based map[J].PLo S Med,2012(11):e1001339.
• [3]Hu R,Lin M,Ye J,et al.Molecular epidemiological investigation of G6PD deficiency by a gene chip among Chinese Hakka of southern Jiangxi province[J].Int J Clin Exp Pathol,2015(11):15013-15018.
• [4]Pretsch W,Charles DJ,Merkle S.X-Linked Glucose-6-Phosphate Dehydrogenase Deficiency in Mus musculus[J].Biochem Genet,1988(1-2):89-103.
• [5]Vargas RA,Sarmiento K,Vasquez IC.Zebrafish(Danio rerio):A Potential Model for Toxinological Studies[J].Zebrafish,2015(5):320-326.
• [6]Wang YP,Zhou LS,Zhao YZ,et al.Regulation of G6PD acetylation by SIRT2 and KAT9 modulates NADPH homeostasis and cell survival during oxidative stress[J].EMBO J,2014(12):1304-1320.
• [7]Westerfield M.The zebrafish book:A guide for the laboratory use of zebrafish(Danio rerio)[M].Univ of Oregon Press,2007:121-134.
• [8]Sandoval JM,Arenas FA,Vásquez CC.Glucose-6-phosphate dehydrogenase protects escherichia coli from tellurite-mediated oxidative stress[J].Plos One,2011(9):e25573.
• [9]Cappellini MD,Fiorelli G.glucose-6-phosphate dehydrogenase deficiency[J].Lancet,2008(371):64-74.
• [10]Rochford R,Ohrt C,Baresel PC,et al.Humanized mouse model of glucose-6-phosphate deh ydrogenase deficiency for in vivo assessment of hemolytic toxicity[J].PASN,2013(43):17486-17491.
• [11]Goessling W,North TE.Hematopoietic stem cell development:using the zebrafish to identify the signaling networks and physical forces regulating hematopoiesis.Methods in cell biology[J].Methods Cell Biol,2011(105):117-136.
• [12]Kanungo J,Cuevas E,Ali SF,et al.Zebrafish model in drug safety assessment.Current pharmaceutical design[J].Curr Pharm Des,2014(34):5416-5429.
• [13]Sabaawy HE,Azuma M,Embree LJ,et al.TEL-AML1transgenic zebrafish model of precursor B cell acute lymphoblastic leukemia[J].PNSA,2006(41):15166-15171.
• [14]Gutierrez A,Grebliunaite R,Feng H,et al.Pten mediates Myc oncogene dependence in a conditional zebrafish model of T cell acute lymphoblastic leukemia[J].J Exp Med,2011(8):1595-1603.
• [15]Von Seidlein L,Auburn S,Espino F,et al.Review of key knowledge gaps in glucose-6-phosp hate dehydrogenase deficiency detection with regard to the safe clinical deployment of 8-aminoquinoline treatment regimens:a workshop report[J].Malar J,2013(12):112.

文章评论(Comment)：