Comparative Effects of Liensinine and Neferine on the Human Ether-a-go-go-related Gene Potassium Channel and Pharmacological Activity Analysis
Comparative Effects of Liensinine and Neferine on the Human Ether-a-go-go-related Gene Potassium Channel and Pharmacological Activity Analysis
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莲心碱与莲心碱对人ether-a-go-go相关基因钾通道的影响比较及药理活性分析
DOI：
10.1159/000338497
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发表时间：
2012-01-01
影响因子：
--
通讯作者：
Li, Bao-Xin
中科院分区：
文献类型：
--
作者：
Dong, Zeng-Xiang;Zhao, Xin;Li, Bao-Xin
莲心碱和甲基莲心碱均为异喹啉类生物碱，具有抗室性心律失常作用。人类ether-a-go-go-related基因（hERG）参与心脏动作电位的复极。我们研究了莲心碱和甲基莲心碱对hERG通道生物物理特性的影响及其构效关系。采用全细胞膜片钳技术、免疫印迹分析和免疫荧光实验检测莲心碱和甲基莲心碱对稳定转染HEK 293细胞hERG通道的影响。采用高效液相色谱法测定莲心碱和甲基莲心碱在大鼠体内的药代动力学和组织分布。莲心碱和甲基莲心碱均能剂量依赖性地降低电流幅值。莲心碱使hERG尾电流从对照组的70.3 ± 6.3 pA/pF降低到1 μ M组的56.7 ± 2.8 pA/pF、3 μ M组的53.0 ± 2.3 pA/pF和30 μ M组的17.8 ± 0.7 pA/pF;甲基莲心碱处理的细胞的相应电流密度分别为41.9 +/-3.1pA/pF，32.3 +/-3.1pA/pF和16.2 +/-0.6pA/pF。甲基莲心碱对hERG通道开放和失活状态均有亲和力，莲心碱仅与开放状态的hERG通道结合。莲心碱和甲基莲心碱对F656 V或Y 652 A突变型通道hERG电流的抑制作用减弱。甲基莲心碱在大鼠体内的分布比莲心碱快，且浓度高于莲心碱。莲心碱和甲基莲心碱对hERG通道的产生和表达均无影响。总之，甲基莲心碱在低浓度下是比莲心碱更有效的hERG通道阻断剂（
Liensinine and neferine, a kind of isoquinoline alkaloid, can antagonize the ventricular arrhythmias. The human ether-a-go-go-related gene (hERG) is involved in repolarization of cardiac action potential. We investigated the effects of liensinine and neferine on the biophysical properties of hERG channel and the underlying structure-activity relationships. The effects of liensinine and neferine were examined on the hERG channels in the stable transfected HEK293 cells using a whole-cell patch clamp technique, western blot analysis and immunofluorescence experiment. The pharmacokinetics and tissue distribution determination of liensinine and neferine in rats were determined by a validated RP-HPLC method. Liensinine and neferine induced decrease of current amplitude in dose-dependent. Liensinine reduced hERG tail current from 70.3 +/- 6.3 pA/pF in control group to 56.7 +/- 2.8 pA/pF in the 1 mu M group, 53.0 +/- 2.3 pA/pF (3 mu M) and 17.8 +/- 0.7 pA/pF (30 mu M); the corresponding current densities of neferine-treated cells were 41.9 +/- 3.1 pA/pF, 32.3 +/- 3.1 pA/pF and 16.2 +/- 0.6 pA/pF, respectively. Neferine had binding affinity for the open and inactivated state of hERG channel, liensinine only bound to the open state. The inhibitory effects of liensinine and neferine on hERG current were attenuated in the F656V or Y652A mutant channels. Neferine distributed more quickly than liensinine in rats, which was found to be in higher concentration than liensinine. Both liensinine and neferine had no effect on the generation and expression of hERG channels. In conclusion, neferine is a more potent blocker of hERG channels than liensinine at low concentration (