Ke Qingen, Xiao Yong-Fu, Bradbury J Alyce, Graves Joan P, Degraff Laura M, Seubert John M, Zeldin Darryl C
National Institutes of Health/NIEHS, 111 T. W. Alexander Drive, Building 101, Room D236, Research Triangle Park, NC 27709, USA.
Mol Pharmacol. 2007 Oct;72(4):1063-73. doi: 10.1124/mol.107.035881. Epub 2007 Jul 24.
CYP2J2 is abundant in cardiac tissue and active in the biosynthesis of eicosanoids such as epoxyeicosatrienoic acids (EETs). To determine the effects of CYP2J2 and its eicosanoid products in the heart, we characterized the electrophysiology of single cardiomyocytes isolated from adult transgenic (Tr) mice with cardiac-specific overexpression of CYP2J2. CYP2J2 Tr cardiomyocytes had a shortened action potential. At 90% repolarization, the action potential duration (APD) was 30.6 +/- 3.0 ms (n = 22) in wild-type (Wt) cells and 20.2 +/- 2.3 ms (n = 19) in CYP2J2 Tr cells (p < 0.005). This shortening was probably due to enhanced maximal peak transient outward K(+) currents (I(to,peak)), which were 38.6 +/- 2.8 and 54.4 +/- 4.9 pA/pF in Wt and CYP2J2 Tr cells, respectively (p < 0.05). In contrast, the late portion of the transient outward K(+) current (I(to,280ms)), the slowly inactivating outward K(+) current (I(K,slow)), and the voltage-gated Na(+) current (I(Na)) were not significantly altered in CYP2J2 Tr cells. N-Methylsulphonyl-6-(2-proparglyloxy-phenyl)hexanamide (MS-PPOH), a specific inhibitor of EET biosynthesis, significantly reduced I(to,peak) and increased APD in CYP2J2 Tr cardiomyocytes but not in Wt cells. Intracellular dialysis with a monoclonal antibody against CYP2J2 also significantly reduced I(to,peak) and increased APD in CYP2J2 Tr cardiomyocytes. Addition of 11,12-EET or 8-bromo-cAMP significantly reversed the MS-PPOH- or monoclonal antibody-induced changes in I(to,peak) and APD in CYP2J2 Tr cells. Together, our data demonstrate that shortening of the action potential in CYP2J2 Tr cardiomyocytes is associated with enhanced I(to,peak) via an EET-dependent, cAMP-mediated mechanism.
细胞色素P450 2J2(CYP2J2)在心脏组织中含量丰富,且在类花生酸生物合成中发挥作用,如环氧二十碳三烯酸(EETs)。为了确定CYP2J2及其类花生酸产物在心脏中的作用,我们对从成年转基因(Tr)小鼠分离的单个心肌细胞的电生理学特性进行了研究,这些小鼠心脏特异性过表达CYP2J2。CYP2J2转基因心肌细胞的动作电位缩短。在复极化90%时,野生型(Wt)细胞的动作电位持续时间(APD)为30.6±3.0毫秒(n = 22),而CYP2J2转基因细胞为20.2±2.3毫秒(n = 19)(p < 0.005)。这种缩短可能是由于最大峰值瞬时外向钾离子电流(I(to,peak))增强,在Wt细胞和CYP2J2转基因细胞中,该电流分别为38.6±2.8和54.4±4.9皮安/皮法(p < 0.05)。相比之下,CYP2J2转基因细胞中瞬时外向钾离子电流的后期部分(I(to,280ms))、缓慢失活的外向钾离子电流(I(K,slow))和电压门控钠离子电流(I(Na))没有显著改变。N - 甲基磺酰基 - 6 - (2 - 炔丙氧基 - 苯基)己酰胺(MS - PPOH),一种EET生物合成的特异性抑制剂,可显著降低CYP2J2转基因心肌细胞中的I(to,peak)并增加APD,但对Wt细胞无此作用。用抗CYP2J2单克隆抗体进行细胞内透析也可显著降低CYP2J2转基因心肌细胞中的I(to,peak)并增加APD。添加11,12 - EET或8 - 溴 - cAMP可显著逆转MS - PPOH或单克隆抗体诱导的CYP2J2转基因细胞中I(to,peak)和APD的变化。总之,我们的数据表明,CYP2J2转基因心肌细胞中动作电位的缩短与通过EET依赖性、cAMP介导的机制增强的I(to,peak)有关。