目的:构建miR-22心肌特异转基因斑马鱼系,在体评估miR-22对于心肌肥厚的作用。方法:构建pTol2-CMLC2-miR-22-IRES-EGFP表达载体。通过显微注射的方法将tol2重组质粒于一细胞期注射入斑马鱼受精卵胚胎中,荧光筛选获得心肌特异表达绿色荧光的斑马鱼胚胎,并稳定表达传代。然后对稳定传代的成年斑马鱼心脏进行心肌肥厚及心功能的检测。结果:成功建立了miR-22心肌特异转基因斑马鱼系,通过定量PCR确定心肌中miR-22表达升高,荧光显微镜观察发现斑马鱼心肌出现绿色荧光。miR-22心脏特异过表达的转基因鱼系的成年鱼与野生对照组相比,出现了心肌肥厚的现象,心肌肥厚分子标志物nppa、myh7明显升高。斑马鱼心脏病理切片结果同样显示出miR-22心肌特异转基因斑马鱼出现了心肌肥厚的现象。结论:成功构建了miR-22心肌特异转基因斑马鱼,为研究心肌中miR-22的生物学功能提供了重要的工具,并证明miR-22心脏特异过表达会引起斑马鱼心肌肥厚。 OBJECTIVE: To construct a miR-22 cardiac-specific transgenic zebrafish line to assess the effect of miR-22 on cardiac hypertrophy in vivo. Methods: The pTol2-CMLC2-miR-22-IRES-EGFP expression vector was constructed. The recombinant plasmids tol2 were injected into zebrafish zygotes embryos at a single cell stage by microinjection, and zebrafish embryos with green fluorescence were obtained by fluorescence screening. Then the stable passage of adult zebrafish hearts for cardiac hypertrophy and cardiac function testing. Results: The miR-22 cardiac-specific transgenic zebrafish line was established successfully. The expression of miR-22 in myocardium was confirmed by quantitative PCR. The green fluorescence of zebrafish myocardium was observed by fluorescence microscopy. Compared with the wild-type control group, cardiac hypertrophy was observed in adult fish of transgenic fish lines with miR-22 cardiac-specific overexpression. The markers of cardiac hypertrophy, nppa and myh7, were significantly increased. Zebrafish cardiac biopsies also showed cardiac hypertrophy in miR-22 cardiac-specific transgenic zebrafish. CONCLUSION: The miR-22 cardiac-specific transgenic zebrafish was successfully constructed, which provided an important tool for studying the biological function of miR-22 in myocardium and demonstrated that cardiac-specific over-expression of miR-22 could cause cardiac hypertrophy in zebrafish.