决奈达隆对新生大鼠心室肌细胞HCN通道mRNA和蛋白表达的影响  

Effect of dronedarone on HCN channel mRNA and protein expression in neonatal rat ventricular myocytes

作　　者：陈琳琳[1] 范新荣[1] 李涛 李光[1] 李妙龄[1] 欧贤红[1] 兰欢[1] 黄梦颖[1] 曾晓荣[1] Chen Linlin Fan Xinrong Li Tao Li Guang Li Miaoling Ou Xianhong Lan Huan Huang Mengying Zeng Xiaorong(Key Laboratory of Medical Electrophysiology of Ministry of Education ,Collaborative Innovation Center for Prevention and Treatment of Cardiovascular Disease/Institute of Cardiovascular Research, Southwest Medical University, Luzhou , Sichuan 646000, Chin)

机构地区：[1]西南医科大学医学电生理学省部教育部重点实验室/四川省心血管疾病防治协同创新中心,四川泸州646000

出　　处：《重庆医学》2017年第9期1153-1155,共3页Chongqing medicine

基　　金：国家自然科学基金资助项目(81500259)

摘　　要：目的通过检测新生大鼠心室肌细胞在给予药物决奈达隆前后超极化激活环核苷酸门控阳离子通道(HCN通道)mRNA和蛋白水平的变化,探讨决奈达隆对HCN通道表达的影响。方法分离新生SD大鼠心室肌,Ⅱ型胶原酶消化,通过差速贴壁分离法收集获得单一的心室肌细胞。并根据浓度(0.1、0.5、1.0、5.0、10.0、20.0μmol/L的决奈达隆对细胞进行处理48h)和时间(浓度为10μmol/L的决奈达隆对细胞进行1、6、12、24、48h的处理)梯度分组。采用实时荧光定量PCR法和Western blot检测HCN2、HCN4通道mRNA水平和蛋白水平。结果浓度梯度组和时间梯度组的HCN2 mRNA和HCN4 mRNA表达水平均低于对照组(P<0.05);与对照组比较,10μmol/L决奈达隆处理后12h组的蛋白水平明显下调(P<0.01)。结论决奈达隆可抑制HCN2、HCN4通道mRNA和蛋白的表达,且作用呈浓度依赖性,在给药后12h达到最大作用。Objective To explore the effect of dronedaronel on hyperpolarization-activated cyclic-nucleotide-gated（HCN） channel expression by detecting the change of HCN channel mRNA and protein level before and after giving dronedarone in neonatal rat ventricular myocytes. Methods Neonatal rat ventricular myocytes were separated and digested by type il collagenase,and then siiagle ventricular myocytes were collected through differential sticking wall separation method. According to the concentrations （0.1,0.5,1.0,5.0,10.0,20.0 ~tmol/L of dronedaronel for treating myocytes for 48 h ） and time（10 μmol/L of dronedaronel for treating myocytes for 1,6,12,24,48 h）the gradient grouping was conducted. The levels of HCN2 and HCN4 channel mRNA and protein level were determined by real-time PCR and Western blot. Results The HCN2 mRNA and HCN4 mRNA expression levels in concentration gradient group and time gradient group were lower than those in the control group（P〈0.05） ;compared with the control group,the protein level in the 10 umol/L dronedaronel treatment for 12 h group was significantly down-regulated（P〈 0.01）. Conclusion Dronedaronel could inhibit the expression of HCN2/HCN4 channel mRNA and protein,moreover its action shows the concentration dependency and reaches the maximum at 12 h after medication.

关 键 词：决奈达隆 超极化激活环核苷酸门控阳离子通道 RNA 信使 蛋白质类 

分 类 号：R96[医药卫生—药理学]