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目的 蛋白激酶Cη(PKCη)转基因对心脏表型的影响。 方法表达低水平 (Tg PKCη L)和高水平 (Tg PKCη H)靶心脏野生型PKCη的转基因小鼠通过标准技术被建立。Tg PKCη L、Tg PKCη H阳性鼠通过PCR和Southern印迹鉴定 ,转基因阴性鼠作为研究对照。用非开胸经颈总动脉插管的显微外科技术测血液动力学指标和评价小鼠左室收缩功能。Western免疫印迹分析α 骨骼肌肌动蛋白 (α |SkeletalMuscleActin)和PKCη表达水平。苏木精染色进行心脏组织学分析。 结果与对照鼠相比Tg PKCη L、Tg PKCη H鼠基线左室最大收缩压、左室收缩压、最大收缩速率 (dp/dt)、 dp/dt、左室舒张末压、舒张压均无明显变化 (均P >0 0 5 ) ,异丙肾上腺素激发后左室dp/dt剂量依赖的升高无明显抑制 (均P >0 0 5 )。此外 ,Tg PKCη H、Tg PKCη L鼠心脏 /体重比、心肌α 肌动蛋白表达无显著变化 ,(P >0 0 5 ) ,心肌组织学分析表现了正常的心肌细胞形态。结论 低水平和高水平PKCη的转基因过渡表达未导致心肌肥厚、心衰表型的出现。用非开胸经颈总动脉插管的显微外科方法能敏感、准确地评价小鼠左室收缩功能。
Effect of PKCn gene transfection on cardiac phenotype. Methods Transgenic mice expressing a low level (Tg PKCn L) and a high level (Tg PKCn H) target-cardiac wild-type PKCη were established by standard techniques. Tg PKCn L, Tg PKCn H positive mice were identified by PCR and Southern blot, and transgenic negative mice served as study controls. Hemodynamic parameters were measured by microsurgical technique without intubation through common carotid artery and left ventricular systolic function was evaluated. Western immunoblotting was used to analyze α-skeletal actin (α | | Skeletal MuscleActin) and PKCη expression levels. Hematoxylin staining for cardiac histological analysis. Results Compared with the control rats, the baseline systolic blood pressure, left ventricular systolic pressure, dp / dt, dp / dt, left ventricular end-diastolic pressure and diastolic blood pressure of Tg PKCn L, Tg PKCn H rats were not significantly (All P> 0.05). The isoproterenol-induced left ventricular dp / dt dose-dependent increase was not significantly inhibited (all P> 0.05). In addition, there was no significant change in cardiac PKCη H, Tg PKCη L rat cardiac / body weight ratio and myocardial a-actin expression (P> 0.05). Myocardial histological analysis showed normal cardiomyocyte morphology. Conclusion Low and high levels of PKCη transgene expression did not lead to cardiac hypertrophy, heart failure phenotype. Microsurgery with non-open thoracic carotid artery cannulation can evaluate the left ventricular systolic function in mice sensitively and accurately.