dos Remedios C G, Berry D A, Carter L K, Coumans J V, Heinke M Y, Kiessling P C, Seeto R K, Thorvaldson T, Trahair T, Yeoh T, Yao M, Gunning P W, Hardeman E, Humphery-Smith I, Naidoo D, Keogh A
Muscle Research Unit, Department of Anatomy, The University of Sydney, Australia.
Electrophoresis. 1996 Jan;17(1):235-8. doi: 10.1002/elps.1150170140.
A variety of electrophoretic techniques were used to search for potential causes of human dilated cardiomyopathy (DCM). Northern blots were used to quantify alpha-cardiac and alpha-skeletal muscle actins, and beta-myosin heavy chain mRNAs which are the predominant expressed isoform species. We found a wide range of mRNA levels expressed in both DCM and nondiseased (ND) samples of left ventricles. However, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) gels of the same heart samples revealed a stable and constant ratio of actin and myosin. Dystrophin deficiency might account for the DCM symptoms and so dystrophin levels of DCM and ND samples were evaluated using Western blots probed with monoclonal antibodies for the N-, C- and mid-rod portions of this protein. We found that dystrophin levels were constant in all 29 DCM and 5 ND samples suggesting that dystrophin deficiency is probably not a contributing cause. We explored the possibility that terminal failure may be due to an apoptotic-like event in the cardiomyocytes. Zymograms of DCM and ND samples revealed a significant increase in DNase I activity in the DCM group compared to the ND samples. These data raise the possibility that end-stage failure may be associated with apoptosis.
采用多种电泳技术探寻人类扩张型心肌病（DCM）的潜在病因。利用Northern印迹法对α-心肌肌动蛋白、α-骨骼肌肌动蛋白以及β-肌球蛋白重链mRNA（这些是主要表达的同工型种类）进行定量分析。我们发现，在左心室的DCM样本和非患病（ND）样本中均表达出了广泛的mRNA水平。然而，相同心脏样本的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）凝胶显示，肌动蛋白和肌球蛋白的比例稳定且恒定。肌营养不良蛋白缺乏可能是DCM症状的原因，因此，使用针对该蛋白N端、C端和中间杆状部分的单克隆抗体进行Western印迹分析，评估DCM样本和ND样本中的肌营养不良蛋白水平。我们发现，在所有29个DCM样本和5个ND样本中，肌营养不良蛋白水平均保持恒定，这表明肌营养不良蛋白缺乏可能并非致病原因。我们探讨了终末期衰竭可能是由于心肌细胞中类似凋亡的事件所致的可能性。DCM样本和ND样本的酶谱分析显示，与ND样本相比，DCM组的脱氧核糖核酸酶I活性显著增加。这些数据表明终末期衰竭可能与细胞凋亡有关。
