Lai Ling-Ping, Lin Jiunn-Lee, Lin Chich-Sheng, Yeh Huei-Ming, Tsay Yeou-Guang, Lee Chwen-Fang, Lee Hsiao-Hui, Chang Zee-Fen, Hwang Juey-Jen, Su Ming-Jai, Tseng Yung-Zu, Huang Shoei K Stephen
Institute of Pharmacology, National Taiwan University, Taipei, Taiwan.
J Cardiovasc Electrophysiol. 2004 Feb;15(2):214-23. doi: 10.1046/j.1540-8167.2004.03423.x.
Functional and structural changes of atrial tissue occur during the natural course of atrial fibrillation (AF), and these changes may contribute to further AF. We investigated the changes in AF tissue using cDNA microarray and two-dimensional protein electrophoresis techniques.
We established a porcine model of AF by rapid right atrial appendage pacing at a rate of 600/min. Atrial tissue was obtained after rapid atrial depolarization for 6 weeks. Microarrays containing 6,035 cDNA clones were used to evaluate the alterations of mRNA. Two-dimensional protein electrophoresis was performed to compare protein patterns. In cDNA microarray studies, we identified 387 genes with significant change in the left atrium and 81 genes in the right atrium. Among the genes, the ventricular isoform of the myosin regulatory light chain (MLC-2V) showed the greatest fold of change (9.4 and 7.3 in the left and right atrium, respectively). In protein electrophoresis, the expression levels of three protein spots spanning from 18 to 20 kDa in the acidic region (PI 4.5-5.0) were specifically elevated in the AF group. Interestingly, through tandem mass spectrometric analysis, these three spots were identified as MLC-2V. Thus, MLC-2V expression at the mRNA and protein levels corresponded well, and both indicated a significant increase in AF.
Both cDNA microarray and two-dimensional polyacrylamide protein electrophoresis studies revealed characteristic changes in AF tissue. We demonstrated the reprogramming of myosin regulatory light chain isoform composition, with a significant increase of its ventricular isoform (MLC-2V).
心房颤动（AF）自然病程中会发生心房组织的功能和结构改变，这些改变可能促使房颤进一步发展。我们采用cDNA微阵列和二维蛋白质电泳技术研究房颤组织的变化。
我们通过以600次/分钟的速率快速右心耳起搏建立了猪房颤模型。快速心房去极化6周后获取心房组织。使用包含6035个cDNA克隆的微阵列评估mRNA的改变。进行二维蛋白质电泳以比较蛋白质图谱。在cDNA微阵列研究中，我们在左心房中鉴定出387个有显著变化的基因，在右心房中鉴定出81个基因。其中，肌球蛋白调节轻链（MLC-2V）的心室异构体变化倍数最大（左心房和右心房分别为9.4和7.3）。在蛋白质电泳中，AF组酸性区域（PI 4.5 - 5.0）中分子量在18至20 kDa之间的三个蛋白点的表达水平特异性升高。有趣的是，通过串联质谱分析，这三个点被鉴定为MLC-2V。因此，MLC-2V在mRNA和蛋白质水平的表达良好对应，且均表明在房颤中显著增加。
cDNA微阵列和二维聚丙烯酰胺蛋白质电泳研究均揭示了房颤组织的特征性变化。我们证明了肌球蛋白调节轻链异构体组成的重新编程，其心室异构体（MLC-2V）显著增加。
