摘要
目的 探索环状RNA mmu_circ_0005019调控小电导钙激活钾(small-conductance calcium-activated potassium, SK)通道蛋白亚基SK3编码基因Kcnn3的表达,以及对小电导钙激活钾通道电流(IK,Ca)和动作电位时程(action potential duration, APD)的影响。方法 在小鼠HL-1细胞中分别构建mmu_circ_0005019过表达和干扰模型,分为过表达组(n=3)、空质粒组(n=3)、干扰1组(n=3)、干扰2组(n=3)、对照组(n=3),通过RT-qPCR、Western blot分析mmu_circ_0005019调节Kcnn3表达的分子机制,应用膜片钳技术电流钳模式记录全细胞的IK,Ca,并用电压钳模式记录APD。结果 成功构建了环状RNA mmu_circ_0005019过表达和干扰的HL-1细胞模型。过表达组的Kcnn3基因表达与空质粒组相比明显上调(P<0.05);干扰1组和干扰2组的Kcnn3基因表达与对照组相比均明显下调(P<0.05)。电生理发现过表达mmu_circ_0005019增加了HL-1细胞IK,Ca电流密度,APD显著缩短;相反,干扰mmu_circ_0005019减少了IK,Ca电流密度,APD显著延长。结论 mmu_circ_0005019可以上调小鼠HL-1细胞Kcnn3的表达水平,从而改变IK,Ca和APD,提示环状RNA mmu_circ_0005019可能在房颤发生中起到促进作用。
Objective To investigate the effect of mmu_circ_0005019 on regulating Kcnn3 expression,small conductance calcium-activated potassium current(I K,Ca)which is generated by the small conductance calcium-activated potassium(SK)channel that is partly coded by Kcnn3,and action potential duration(APD).Methods We transfected pcDNA3.1-mmu_circ_0005019 expressing vectors and 2 independent small interfering RNAs into HL-1 cells to establish gain-and loss-of-expression cell models,respectively.The transfected cells were divided into overexpression group(n=3),empty vector group(n=3),interference 1 group(n=3),interference 2 group(n=3),and blank control group(n=3).RT-qPCR and Western blotting were used to determine the expression of Kcnn3.Whole-cell patch clamp technology was performed to record the I K,Ca and APD.The effects of mmu_circ_0005019 on Kcnn3 expression,I K,Ca and APD were observed.Results Gain-and loss-of-expression cell models were established successfully.The expression of Kcnn3 and I K,Ca current density significantly increased,and the APD was shortened in overexpression group compared with empty vector group(P<0.05).The expression of Kcnn3 and I K,Ca current density were significantly decreased,and the APD was prolonged in interference groups compared with blank control group(P<0.05).Conclusion Mmu_circ_0005019 promotes the expression of Kcnn3 and its coding channel protein,which regulates I K,Ca and APD in mouse cardiomyocytes,and may play a promoting role in the occurrence of atrial fibrillation.
作者
杨岚清
邬娜
陈鹏慧
袁志权
李成英
吴龙
钟理
李亚斐
YANG Lanqing;WU Na;CHEN Penghui;YUAN Zhiquan;LI Chengying;WU Long;ZHONG Li;LI Yafei(Department of Epidemiology,College of Military Preventive Medicine,College of Basic Medical Sciences,Army Medical University(Third Military Medical University),Chongqing,400038;Department of Neurobiology,College of Basic Medical Sciences,Army Medical University(Third Military Medical University),Chongqing,400038;Cardiovascular Disease Center,the Third Affiliated Hospital of Chongqing Medical University,Chongqing,401120,China)
出处
《陆军军医大学学报》
CSCD
北大核心
2024年第2期100-109,共10页
Journal of Army Medical University
基金
国家自然科学基金面上项目(82073649)。
关键词
环状RNA
心肌细胞
小电导钙激活钾通道
动作电位时程
膜片钳
circular RNA
cardiomyocyte
small conductance calcium-activated potassium current
action potential duration
patch clamp
