房颤冷冻球囊消融术对调控编码超速激活延迟整流钾通道蛋白的影响

Effect of Cryo-balloon Ablation of Atrial Fibrillation on Regulating and Encoding Delayed Rectifier Potassium Channel Protein

目的探讨房颤冷冻球囊消融术对调控编码超速激活延迟整流钾通道蛋白(KCNA5)miRNA的影响,寻找可能的miRNA调控干预靶点,为房颤的治疗提供新思路。方法选取2017年1月~2019年10月新疆维吾尔自治区人民医院接受冷冻球囊消融术后随访3个月无复发患者作为研究对象,分别于术前、术后抽取患者外周血进行全基因组测序,将术前测序作为对照组,术后测序作为试验组,比较两组miRNA表达差异,并通过mirbase、miranda、targetscan3个数据库进行靶基因分析,对存在差异的miRNA进行实时定量逆转录聚合酶链式反应和蛋白质印迹法验证。结果全基因组测序显示,两组miRNA表达比较,差异有统计学意义(P<0.05),其中has-miR-134-5p等15个miRNAs术后下调,has-miR-1255a等6个miRNAs术后上调;调控超速激活延迟整流钾通道蛋白的miRNAs经过逆转录聚合酶链式反应验证提示试验组KCNA5表达较对照组下调[(0.398±0.060)vs(1.000±0.035)],差异有统计学意义(P<0.05),且与Western Blot检测趋势一致。结论冷冻球囊消融术影响了房颤离子流的动态平衡,调控编码超速激活延迟整流钾通道蛋白的hsa-miR-4433b-3p等miRNAs有可能为房颤新的生物标识物和潜在的治疗靶标。

Objective To investigate the effect of cryo-balloon ablation of atrial fibrillation on the regulation of miRNAs encoding super-activated delayed rectifier potassium channel protein(KCNA5),and to find possible miRNA regulatory intervention targets,and to provide new ideas for the treatment of atrial fibrillation.Methods From January 2017 to October 2019,patients with no recurrence followed up for 3 months after cryo-balloon ablation in Xinjiang Uygur Autonomous Region People's Hospital were selected as the research subjects.Peripheral blood was drawn from patients before and after surgery for whole genome sequencing.The preoperative sequencing was used as the control group,and the postoperative sequencing was used as the test group.The differences in miRNA expression between the two groups were compared,and target gene analysis was performed through the three databases of mirbase,miranda,and targetscan.Perform real-time quantitative reverse transcription polymerase chain reaction and Western blotting to verify the different miRNAs.Results Whole-genome sequencing showed that there was a statistically significant difference in the expression of miRNA between the two groups(P<0.05).Among them,15 miRNAs such as has-miR-134-5p were down-regulated after surgery,and 6 miRNAs,such as has-miR-1255a,were up-regulated after surgery;miRNAs that regulate the super-rapid activation of delayed rectifier potassium channel protein were verified by reverse transcription polymerase chain reaction.The expression of KCNA5 in the group was compared with that in the control group[(0.398±0.060)vs(1.000±0.035)],the difference was statistically significant(P<0.05),and it was consistent with the Western Blot detection trend.Conclusion Cryoballoon ablation affects the dynamic balance of atrial fibrillation ion currents.The regulation of miRNAs such as hsa-miR-4433b-3p,which encodes hyperactive delayed rectifier potassium channel protein,may be a new biomarker and potential therapeutic target for atrial fibrillation.