CYP3A65及上游核受体基因PXR和AHR2在斑马鱼生长发育阶段的表达

Expression of metabolic enzyme CYP3A65 and its upstream nuclear receptor PXR and AHR2 genes during zebrafish(Danio rerio) developmental stages

为了选择处于最佳生长阶段的斑马鱼用于药物代谢研究,本实验考察不同生长阶段斑马鱼全鱼、肝脏和肠道组织中CYP3A65及其上游核受体基因(PXR、AHR2)mRNA表达水平的变化规律。实验测定CYP3A65、PXR和AHR2相对表达量,并通过比较核受体基因与代谢酶基因mRNA之间的相关性来证实核受体基因的调控作用。结果显示,144 hpf和75 dpf两个生长阶段的斑马鱼中CYP3A65、PXR和AHR2 mRNA表达量最高;斑马鱼生长阶段中代谢酶基因和核受体基因的表达趋势一致,且受肝脏、肠道组织占全鱼比重的影响。研究表明,幼鱼期144 hpf斑马鱼及稚鱼期75 dpf的斑马鱼最适用于CYP3A65代谢研究;CYP3A65与PXR、CYP3A65与AHR2的mRNA表达之间有显著的相关性,与AHR2的相关性更显著。

Drug metabolizing enzyme CYP3A65 is the only member of Danio rerio CYP3A subfamily.Our study was conducted to select appropriate D.rerio developmental stage,by investigating the expression of CYP3A65 and its its upstream nuclear receptor(PXR and AHR2)during developmental stages.The zebrafish in different growth stages were raised,and the whole fish tissues,liver tissues and intestinal tissues were taken.The relative expression levels of CYP3A65,PXR and AHR2 were measured,and the correlation between nuclear receptors and metabolic enzyme mRNA was compared to confirm the regulating effect of nuclear receptors.In whole fish,liver and intestine tissues,the mRNA levels of CYP3A65,PXR and AHR2 were the highest in 144 hpf larvae and 75 dpf juvenile D.rerio.The change trends of metabolic enzyme and nuclear receptors gene expression were consistent,and affected by the proportion of liver and intestine relative to whole fish.The 144 hpf and 75 dpf D.rerio may be in the juvenile stage and throughout the growth phase,and are most suitable for CYP3A65 metabolism studies.There were significant correlations between CYP3A65 and PXR、CYP3A65 and AHR2 mRNA expression,and correlation between CYP3A65 and AHR2 was better.In our later work,the enzyme activity and protein expression will be further investigated during D.rerio development,which would provide more information for the model organism D.rerio.