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目的建立心脏特异表达人多巴胺D5受体突变基因F173L(D5F173L)的转基因小鼠,利用该转基因动物模型来研究多巴胺D5受体在心脏肥大发生中的作用机制。方法利用心脏特异启动子α-MHC构建转基因表达载体,显微注射法建立心脏特异表达人多巴胺D5F173L(α-MHC-D5F173L)的转基因小鼠,PCR鉴定转基因小鼠的基因型,Western blot检测多巴胺D5受体在心脏组织中的表达,心脏超声检测转基因小鼠及野生小鼠的心脏结构和功能。光学显微镜检查α-MHC-D5F173L转基因小鼠心脏的病理改变。结果建立了α-MHC-D5F173L转基因小鼠。在3月龄,与野生型小鼠比较,α-MHC-D5F173L转基因小鼠心脏收缩期和舒张期左心室内径均增加[收缩期(3·26±0·42)比(2·58±0·23)mm,舒张期(4·28±0·39)比(3·86±0·25)mm,均P<0·05],收缩期和舒张期左心室容积均增大[收缩期(44·97±14·54)比(24·66±5·34)μL,舒张期(83·99±18·42)比(64·83±9·90)μL,均P<0·05],射血分数及短轴缩短率减少[(48·01±8·73)%比(62·18±4·84)%,(24·23±5·15)%比(33·15±3·52)%,均P<0·05]。病理学观察显示,α-MHC-D5F173L转基因小鼠心腔明显大于野生型,心室壁明显变薄,心肌细胞不均匀肥大,心肌间质纤维增多。结论α-MHC-D5F173L转基因小鼠出6现典型扩张型心肌病的表型。
Objective To establish a transgenic mouse with heart-specific expression of human dopamine D5 receptor mutant gene F173L (D5F173L) and to investigate the mechanism of dopamine D5 receptor in cardiac hypertrophy by using the transgenic animal model. Methods The gene expression vector was constructed by cardiac specific promoter α-MHC. The cardiac-specific transgenic mice expressing human dopamine D5F173L (α-MHC-D5F173L) were established by microinjection. The genotypes of transgenic mice were identified by PCR. D5 receptor expression in cardiac tissue, heart tissue and function of cardiac and transgenic mice by cardiac sonography. Pathological changes in the heart of α-MHC-D5F173L transgenic mice were examined by light microscopy. Results α-MHC-D5F173L transgenic mice were established. At 3 months of age, both systolic and diastolic LV diameters increased in the α-MHC-D5F173L transgenic mice (3.26 ± 0.42 vs 2.58 ± 0 · 23 mm, diastolic (4.28 ± 0.39) vs (3.86 ± 0.25) mm, all P <0.05], systolic and diastolic LV volumes increased (44.97 ± 14.54) vs (24.66 ± 5.34) μL and diastolic (83.99 ± 18.42) vs (64.83 ± 9.90) μL, all P <0.05 ], Ejection fraction and shortening rate of short axis [(48.01 ± 8.73)% (62.18 ± 4.84)%, (24.23 ± 5.15)% (33.15 ± 3 · 52)%, all P <0 · 05]. Pathological observations showed that the α-MHC-D5F173L transgenic mice were significantly larger than the wild-type heart chamber, ventricular wall thinning significantly, myocardial cells were uneven hypertrophy, myocardial interstitial fibers increased. Conclusion α-MHC-D5F173L transgenic mice out of 6 cases of typical dilated cardiomyopathy phenotype.