Apocynin预防异丙肾上腺素诱导小鼠阵发性房颤的研究
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摘要
目的探讨Apocynin(APO)预防大剂量异丙肾上腺素(isoproterenol,ISO)诱导小鼠阵发性房颤发生的作用及其可能的作用机制,为临床预防阵发性房颤提供新的治疗思路。方法将18只C57BL/6雄性小鼠随机分为正常组、模型组和治疗组,各6只。以100 mg/kg ISO腹腔注射诱导小鼠发生阵发性房颤,治疗组于造模前0.5 h腹腔注射APO(50 mg/kg)100μL,正常组则给予等体积的溶剂。监测小鼠2 h内动态心电图变化情况,统计分析心房颤动的发生频率、持续时间及期前收缩发生率;2,3,5-氯化三苯基四氮唑(TTC)染色评价心肌缺血梗死情况;检测心房组织中miR-208b、miR-30b以及靶基因KCNN3的表达。结果与模型组比较,治疗组期前收缩及阵发性房颤的发生率、心肌组织缺血梗死面积显著降低(P<0.05);与正常组比较,模型组心房组织中miR-208b、miR-30b显著上调(P<0.05),基因KCNN3的表达显著下调(P<0.05);与模型组比较,治疗组心房组织中miR-208b、miR-30b的表达显著下调,基因KCNN3的表达显著上调(P<0.05)。结论 APO可显著预防ISO诱导小鼠阵发性房颤的发生,其作用机制可能与APO显著调节心房组织中miR-208b、miR-30b的表达,进而调节其靶基因KCNN3的表达有关。
Objective To investigated the effect of Apocynin in preventing against isoproterenol-induced atrial fibrillation in C57BL/6 mice and the underlying mechanisms,in order to offer a new therapy for clinical prevention of paroxysmal atrial fibrillation.Methods C57BL/6 male mice were randomly divided into normal group,model group,and treatment group,6 mice in each groups.Paroxysmal atrial fibrillation was induced in mice by 100 mg/kg ISO intraperitoneal injection.The treatment group received intraperitoneal injection of APO(50 mg/kg) 100 μL for 0.5 h before modeling,and equal volume of solvent was given in the normal group.The dynamic changes of electrocardiogram were monitored for 2 h,and the atrial fibrillation frequency,duration,and the incidence rate of atrial premature beats were statistically analyzed.The heart was then dissected,which was followed by TTC staining as histopathologic stain for identification of myocardial ischemic or infarction.Furthermore,the cardiac expression of miR-208b,miR-30b,and the target gene KCNN3 were analyzed by real-time PCR.Results Compared with the model group,the incidence of presystolic and paroxysmal atrial fibrillation in the treatment group was significantly lower than that in the model group(P<0.05).Compared with the normal group,mir-208b and mir-30b were significantly up-regulated in the atrial tissues of the model group(P<0.05),and gene KCNN3 expression was significantly down-regulated(P<0.05).Compared with the model group,the expressions of mir-208b and mir-30b in the atrial tissues of the treatment group were significantly down-regulated,and the expression of gene KCNN3 was significantly up-regulated(P<0.05).Conclusion APO can significantly prevent the occurrence of isoproterenol-induced atrial fibrillation in mice,and its mechanism may be related to regulate the expression of mir-208b and mir-30b in atrial tissue by APO,so as to regulate the expression of its target gene KCNN3.
Objective To investigated the effect of Apocynin in preventing against isoproterenol-induced atrial fibrillation in C57BL/6 mice and the underlying mechanisms,in order to offer a new therapy for clinical prevention of paroxysmal atrial fibrillation.Methods C57BL/6 male mice were randomly divided into normal group,model group,and treatment group,6 mice in each groups.Paroxysmal atrial fibrillation was induced in mice by 100 mg/kg ISO intraperitoneal injection.The treatment group received intraperitoneal injection of APO(50 mg/kg) 100 μL for 0.5 h before modeling,and equal volume of solvent was given in the normal group.The dynamic changes of electrocardiogram were monitored for 2 h,and the atrial fibrillation frequency,duration,and the incidence rate of atrial premature beats were statistically analyzed.The heart was then dissected,which was followed by TTC staining as histopathologic stain for identification of myocardial ischemic or infarction.Furthermore,the cardiac expression of miR-208b,miR-30b,and the target gene KCNN3 were analyzed by real-time PCR.Results Compared with the model group,the incidence of presystolic and paroxysmal atrial fibrillation in the treatment group was significantly lower than that in the model group(P<0.05).Compared with the normal group,mir-208b and mir-30b were significantly up-regulated in the atrial tissues of the model group(P<0.05),and gene KCNN3 expression was significantly down-regulated(P<0.05).Compared with the model group,the expressions of mir-208b and mir-30b in the atrial tissues of the treatment group were significantly down-regulated,and the expression of gene KCNN3 was significantly up-regulated(P<0.05).Conclusion APO can significantly prevent the occurrence of isoproterenol-induced atrial fibrillation in mice,and its mechanism may be related to regulate the expression of mir-208b and mir-30b in atrial tissue by APO,so as to regulate the expression of its target gene KCNN3.
引文
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[2] MICHAEL A R,SAUMYA D,PABLO Q P,et al.A novel transgenic mouse model of cardiac hypertrophy and atrial fibrillation[J].J Atr Fibrillation,2012,2(9):1-15.
[3] SAJID A S,ASHISH K,VIVEK S,et al.Modified radial v/s biatrial maze for atrial fibrillation in rheumatic valvular heart surgery[J].Indian Heart Journal,2014,66(5):510-516.
[4] LIU L,CUI J,YANG Q,et al.Apocynin attenuates isoproterenol-induced myocardial injury and fibrogenesis[J].Biochemical & Biophysical Research Communications,2014,449(1):55-61.
[5] SCHULTE C,KARAKAS M,ZELLER T.MicroRNAs in cardiovascular disease-clinical application[J].Clinical Chemistry & Laboratory Medicine,2017,55(5):687-704.
[6] HANG P,GUO J,SUN C,et al.MicroRNAs as candidate drug targets for cardiovascular diseases[J].Current Drug Targets,2017,18(4):463-472.
[7] WERNHART S,HALLE M.Atrial fibrillation and long-term sports practice:epidemiology and mechanisms[J].Clinical Research in Cardiology,2015,104(5):369-379.
[8] GUO Y,WANG H,TIAN Y,et al.Time trends of aspirin and warfarin use on stroke and bleeding events in Chinese patients with new-onset atrial fibrillation[J].Chest,2015,148(1):62-72.
[9] QINBO Y,CHENGLIN J,PEIWEI W,et al.MicroRNA-505 identified from patients with essential hypertension impairs endothelial cell migration and tube formation[J].Intenational Journal of Cardiology,2014,177(3):925-934.
[10] 贾成林,吴丹丹,李黎,等.三七皂苷对异丙肾上腺素所诱导小鼠阵发性房颤的预防作用[J].中西医结合心脑血管病杂志,2016,14(24):2883-2887.
[11] BRIASOULIS A,SHARMA S,TELILA T,et al.MicroRNAs in atrial fibrillation[J].Current Medicinal Chemistry,2017,24:1-8.
[12] ZHENJI G,JOHN R,BETHANY C,et al.Nuclear receptor/micro RNA circuitry links muscle fiber type to energy metabolism[J].Clin Invest,2013,12(6):2564-2575.
[13] CALLIS T E,PANDYA K,SEOK H Y,et al.MicroRNA-208a is a regulator of cardiac hypertrophy and conduction in mice[J].Journal of Clinical Investigation,2009,119(9):2772-2786.
[14] HULANICKA M,GARNCARZ M,PARZENIECKA-JAWORSKA M,et al.Plasma miRNAs as potential biomarkers of chronic degenerative valvular disease in Dachshunds[J].BMC Veterinary Research,2014,10(1):205.
[15] NAQVI A R,FORDHAM J B,NARES S.miR-24,miR-30b,and miR-142-3p regulate phagocytosis in myeloid inflammatory cells[J].The Journal of Immunology,2015,194(4):1916-1927.
[16] ELLINOR P T,LUNETTA K L,GLAZER N L,et al.Common variants in KCNN3 are associated with lone atrial fibrillation[J].Nature Genetics,2010,42(3):240-244.
[17] OLESEN M S,JABBARI J,HOLST A G,et al.Screening of KCNN3 in patients with early-onset lone atrial fibrillation[J].Europace,2011,13(7):963-967.
[18] CHANG S H,CHANG S N,HWANG J J,et al.Significant association of rs13376333 in KCNN3 on chromosome 1q21 with atrial fibrillation in a Taiwanese population[J].Circulation Journal,2012,76(1):184-188.
[19] LUO Z,YAN C,ZHANG W,et al.Association between SNP rs13376333 and rs1131820 in the KCNN3 gene and atrial fibrillation in the Chinese Han population[J].Clinical Chemistry and Laboratory Medicine(CCLM),2014,52(12):1867-1873.