[1]陈秀 陈甘潇 张中和 胡丹 夏豪.钙通道突变致早期复极综合征的电生理机制研究[J].心血管病学进展,2021,(5):457-463.[doi:10.16806/j.cnki.issn.1004-3934.2021.05.018]
 CHEN Xiu,CHEN Ganxiao,ZHANG Zhonghe,et al.Electrophysiological Mechanism of Early Repolarization Syndrome Caused by Calcium Channel Mutation[J].Advances in Cardiovascular Diseases,2021,(5):457-463.[doi:10.16806/j.cnki.issn.1004-3934.2021.05.018]
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钙通道突变致早期复极综合征的电生理机制研究()
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《心血管病学进展》[ISSN:51-1187/R/CN:1004-3934]

卷:
期数:
2021年5期
页码:
457-463
栏目:
论著
出版日期:
2021-05-25

文章信息/Info

Title:
Electrophysiological Mechanism of Early Repolarization Syndrome Caused by Calcium Channel Mutation
作者:
陈秀 陈甘潇 张中和 胡丹 夏豪
(武汉大学人民医院心血管内科 武汉大学心血管病研究所 湖北省心脏重点实验室,湖北 武汉 430060)
Author(s):
CHEN Xiu CHEN Ganxiao ZHANG Zhonghe HU Dan XIA Hao
(Department of Cardiology,Renmin Hospital of Wuhan University;Cardiovascular Research Institute of Wuhan University;Hubei Key Laboratory of Cardiology, Wuhan 430060,Hubei,China) (81670304) E-mail: hudan0716@hotmail.comE-mail:xiahao1966@163.com
关键词:
早期复极综合征CACNA1C基因突变蛋白转运
Keywords:
Early repolarization syndrome CACNA1CGene mutationProtein trafficking
DOI:
10.16806/j.cnki.issn.1004-3934.2021.05.018
摘要:
目的 探究CACNA1C-P817S突变对早期复极综合征(early repolarization syndrome,ERS)的影响及其电生理机制。方法 收集ERS先证者外周血,采用二代测序技术,对ERS先证者筛选常见致心肌早期复极的候选基因。构建CACNA1C野生型(WT)和突变型质粒,连同WT-CACNB2b、WT-CACNA2D1质粒,分别共转染入人胚肾293细胞。采用全细胞膜片钳技术检测细胞电生理功能,同时应用共聚焦荧光显微镜检测Cav1.2蛋白的转运功能。结果 基因测序筛选出致ERS的突变基因CACNA1C-P817S,突变使CACNA1C氨基酸序列上817位点的脯氨酸变为丝氨酸。全细胞膜片钳检测结果显示P817S组钙电流(ICa)密度较WT组降低约68.8% [ WT(n=15)vs P817S(n=12):(﹣19.2±1.5)pA/pF vs(﹣6.0±1.7)pA/pF,P<0.001]。L型钙离子通道(LTCC)激活未见明显改变,但失活过程加速[V1/2:WT(n=15)vs P817S(n=12):(﹣30.4±0.75)mV v s(﹣41.6±0.84)mV,P<0.001]。共聚焦检测结果显示,P817S突变可使Cav1.2蛋白转运功能降低(外周荧光强度绝对值:WT vs P817S :(19.6±2.0)MD vs (12.2±1.9)MD,P<0.01,n=4;外周/整体荧光强度百分比:WT vs P817S :(30.2±2.3)% vs (20.9±2.2)%, P<0.01,n=4)。结论 CACNA1C-P817S突变通过降低Cav1.2蛋白的转运功能和改变通道动力学,使ICa发生功能性缺失,提示CACNA1C-P817S突变是ERS的一个明确的致病基因突变。
Abstract:
Objective To study the effect of CACNA1C-P817S mutation on early repolarization syndrome(ERS) and explore the underlying electrophysiological mechanism. Methods After collecting the peripheral blood of the proband ,we sequenced several candidate genes encoding ion channels with next generation sequencing which can contribute to ERS, as reported previously. Wild-type(WT) and mutant plasmids of CACNA1C,together with the WT plasmids of CACNB2b, CACNA2D1 were co-transfected into human embryonic kidney 293 cells respectively. The studies of electrophysiological function were performed by whole patch-clamp,and membrane trafficking of Cav1.2 protein was analyzed through confocal fluorescence microscope. Results Gene sequencing identified the mutat ion of CACNA1C,P817S,at the position 817 of the amino acid sequence leading to amino acid change from proline to serine. The results of whole-cell patch-clamp presented that the density of ICa was reduced approximately by 68.8%,compared to WT[WT(n=15) vs P817S(n=12):(﹣19.2±1.5) pA/pF vs (﹣6.0±1.7)pA/pF,P<0.001]. No significant changes have been seen in steady state activation,but the steady state inactivation was accelerated,compared to WT[V1/2: WT(n=15)vs P817S(n=12):(﹣30.4±0.75)mV vs(﹣41.6±0.84)mV,P<0.001]. The results of confocal fluorescence microscope revealed that Cav1.2 protein membrane trafficking of P817S was decreased [absolute peripheral immunofluorescence intensity:WT vs P817S :(19.6±2.0)MD vs (12.2±1.9)MD,P<0.01,n=4;ratio of peripheral/whole immunofluorescence intensity:WT vs P817S :(30.2±2.3)% vs (20.9±2.2)%,P<0.01,n=4]. Conclusion The CACNA1C-P817S mutation can defect membrane trafficking and change the channel dynamics of Cav1.2 protein,thus inducing a loss-of-function of ICa,suggesting that CACNA1C-P817S is a gene mutation associated to ERS.

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更新日期/Last Update: 2021-06-17