Duff H J, Offord J, West J, Catterall W A
Department of Pharmacology, University of Washington, Seattle 98195.
Mol Pharmacol. 1992 Oct;42(4):570-4.
Previous studies have shown that chronic in vivo treatment with the antiarrhythmic drug mexiletine produces an increase in sodium channel number. We examined whether chronic mexiletine treatment would similarly regulate the level of mRNA encoding the cardiac sodium channel. RNA isolated from cardiac tissue was probed with a 2.5-kilobase cRNA transcribed with T7 RNA polymerase from the clone Na 8.4, which encodes nucleotides 3361-5868 of the alpha subunit of the RIIA sodium channel subtype. Chronic mexiletine treatment produced a 3-fold increase in the level of mRNA encoding sodium channel alpha subunits. Previous studies of cultured skeletal muscle cells had suggested that chronic sodium channel blockade may mediate an increase in sodium channel mRNA by changes in cytosolic Ca2+ concentration. To address this issue, we assessed whether verapamil would also produce up-regulation of the level of mRNA encoding the sodium channel and whether the calcium ionophore A23187 would produce the opposite effect on mRNA level. Verapamil treatment increased sodium channel mRNA level up to 3-fold, whereas in vitro A23187 treatment decreased the mRNA level 5-fold. The combination of verapamil and mexiletine produced no further increase in the mRNA level, compared with that seen with the single agents, suggesting a convergent second messenger pathway for the actions of these two drugs. These data show that the level of mRNA encoding sodium channels is substantially increased during antiarrhythmic drug treatment and suggest that change in cytosolic Ca2+ concentration is the second messenger involved in the regulation of levels of mRNA encoding the alpha subunit of the cardiac sodium channel.
以往的研究表明,用抗心律失常药物美西律进行慢性体内治疗会使钠通道数量增加。我们研究了慢性美西律治疗是否会同样调节编码心脏钠通道的mRNA水平。用T7 RNA聚合酶从克隆Na 8.4转录的2.5千碱基cRNA对从心脏组织分离的RNA进行探针杂交,该克隆编码RIIA钠通道亚型α亚基的核苷酸3361 - 5868。慢性美西律治疗使编码钠通道α亚基的mRNA水平增加了3倍。以往对培养的骨骼肌细胞的研究表明,慢性钠通道阻断可能通过胞质Ca2+浓度的变化介导钠通道mRNA的增加。为了解决这个问题,我们评估了维拉帕米是否也会使编码钠通道的mRNA水平上调,以及钙离子载体A23187是否会对mRNA水平产生相反的影响。维拉帕米治疗使钠通道mRNA水平增加高达3倍,而体外A23187治疗使mRNA水平降低5倍。与单一药物相比,维拉帕米和美西律联合使用并未使mRNA水平进一步增加,这表明这两种药物的作用存在一条趋同的第二信使途径。这些数据表明,在抗心律失常药物治疗期间,编码钠通道的mRNA水平显著增加,并表明胞质Ca2+浓度的变化是参与调节心脏钠通道α亚基编码mRNA水平的第二信使。