氢吗啡酮后处理对大鼠缺血-再灌注心律失常及心肌缝隙连接蛋白43表达的影响
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摘要
目的观察氢吗啡酮后处理对缺血-再灌注大鼠体外心肌再灌注性心律失常及心肌组织缝隙连接蛋白43(Cx43)表达的影响,并探讨其相关机制。方法 SD大鼠24只,随机分为3组:对照组(C组)、缺血-再灌注组(IR组)和氢吗啡酮后处理组(HM组),每组8只。建立Langendorff体外心脏缺血-再灌注模型,连续监测心电图和心率。分析再灌注时心律失常发生情况并评分,采用Western blotting技术检测缺血-再灌注心室肌Cx43和Akt信号通路蛋白的表达。结果 3组大鼠各时点心率组间、组内比较均差异无统计学意义(P>0.05)。再灌注期间,IR组和HM组的室性期前收缩数量和再灌注心律失常评分差异无统计学意义(P>0.05);与IR组比较,HM组发生室速和室颤的大鼠数更少,再灌注时心律失常的持续时间更短(P<0.05)。与C组比较,IR组Akt的表达差异无统计学意义(P>0.05),HM组的Akt表达上调,IR组和HM组心肌Cx43的表达均下调;与IR组比较,HM组心肌Akt和Cx43的表达均上调(P<0.05)。结论氢吗啡酮后处理抑制大鼠心肌缺血-再灌注损伤诱发的心律失常机制与其激活Akt信号通路后上调心肌Cx43表达有关。
Objective To investigate the effect of hydromorphone postconditioning on the reperfusion arrhythmia and the expression of myocardial connexin 43( Cx43) during ischemia-reperfusion in isolated rat hearts and its mechanism.Methods The SD rats( n = 24) were randomly divided into three groups: control group( group C,n = 18),ischemia-reperfusion group( group I/R,n = 8),hydromorphone post-treatment group( group HM,n = 8).The langendorff heart or isolated perfused heart assay was established. The heart rate( HR) and ECG during the whole experiment period were recorded. And the reperfusion arrhythmia during the period of reperfusion were detected by ECG. The expression of Akt and Cx43 protein were detected by Western blotting technique. Results Although the HR at different time points and the reperfusion arrhythmia score of three groups were not statistically significant( P> 0.05),the duration of reperfusion arrhythmia of group HM was significantly shorter than that of group IR,and the incidence of ventricular tachycardia and ventricular fibrillation in group HM were also less than that in group IR( P<0.05). When Akt expression level in group IR was compared with group C,there was no significant differences( P>0.05),but Akt expression level in group HM was obviously increased; Cx43 expression level in group IR and group HM were both significantly reduced. While compared with the group IR,Cx43 expression level in group HM was significantly increased( P< 0. 05). Conclusion The mechanism by which hydromorphone post-treatment inhibits reperfusion arrhythmia induced by myocardial IR is associated with up-regulated expression of myocardial Cx43 after activation of Akt signaling pathway during ischemia-reperfusion in isolated rat hearts.
Objective To investigate the effect of hydromorphone postconditioning on the reperfusion arrhythmia and the expression of myocardial connexin 43( Cx43) during ischemia-reperfusion in isolated rat hearts and its mechanism.Methods The SD rats( n = 24) were randomly divided into three groups: control group( group C,n = 18),ischemia-reperfusion group( group I/R,n = 8),hydromorphone post-treatment group( group HM,n = 8).The langendorff heart or isolated perfused heart assay was established. The heart rate( HR) and ECG during the whole experiment period were recorded. And the reperfusion arrhythmia during the period of reperfusion were detected by ECG. The expression of Akt and Cx43 protein were detected by Western blotting technique. Results Although the HR at different time points and the reperfusion arrhythmia score of three groups were not statistically significant( P> 0.05),the duration of reperfusion arrhythmia of group HM was significantly shorter than that of group IR,and the incidence of ventricular tachycardia and ventricular fibrillation in group HM were also less than that in group IR( P<0.05). When Akt expression level in group IR was compared with group C,there was no significant differences( P>0.05),but Akt expression level in group HM was obviously increased; Cx43 expression level in group IR and group HM were both significantly reduced. While compared with the group IR,Cx43 expression level in group HM was significantly increased( P< 0. 05). Conclusion The mechanism by which hydromorphone post-treatment inhibits reperfusion arrhythmia induced by myocardial IR is associated with up-regulated expression of myocardial Cx43 after activation of Akt signaling pathway during ischemia-reperfusion in isolated rat hearts.
引文
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[19] TU I,YEN H D,CHENG H,et al.Baicalein protects chicken embryonic cardiomyocyte against hypoxia-reoxygenation injury viaμ-andδ-but notκ-opioid receptor signaling[J].Eur J Pharm,2008,588(2/3):251-258.
[2] DESPLANTEZ T.Cardiac Cx43,Cx40 and Cx45 coassembling:involvement of connexins epitopes in formation of hemichannels and Gap junction channels[J]. BMC Cell Biol,2017,18(Suppl 1):3.
[3] MICHELA P,VELIA V,ALDO P,et al.Role of connexin 43in cardiovascular diseases[J]. Eur J Pharm,2015,768(12):71-76.
[4] SCHULZ R,GRGE P M,GRBE A,et al. Connexin 43 is an emerging therapeutic target in ischemia/reperfusion injury,cardioprotection and neuroprotection[J]. Pharmacol Therap,2015,153(9):90-106.
[5] BATRA N,RIQUELME M A,BURRA S,et al.Direct regulation of osteocytic connexin 43 hemichannels through AKT kinase activated by mechanical stimulation[J]. J Biolog Chem,2014,289(15):10582-10591.
[6] DUNN C A,SU V,LAU A F,et al.Activation of Akt,not connexin 43 protein ubiquitination,regulates gap junction stability[J].J Biolog Chem 2012,287(4):2600-2607.
[7] DOU M,WU H,ZHU H,et al.Remifentanil preconditioning protects rat cardiomyocytes against hypoxia-reoxygenation injury via delta-opioid receptor mediated activation of PI3K/Akt and ERK pathways[J]. Eur J Pharm,2016,789(8):395-401.
[8] MASLOV L N,KHALIULIN I,OELTGEN P R,et al.Prospects for creation of cardioprotective and antiarrhythmic drugs based on opioid receptor agonists[J]. Med Res Rev,2016,36(5):871-923.
[9] JOHN P HEADRICK L E S H.Opioid receptors and cardioprotection!‘opioidergic conditioning’of the heart[J].Br J Pharm,2015,2(17):2026-2050.
[10] MURRAY A,HAGEN N A.Hydromorphone[J]. J Pain Symp Mana,2005,29(5):57-66.
[11]王小晓,王幼平,余海滨,等.大鼠Langendorff离体心脏灌流模型的制备经验及其影响因素分析[J].中药药理与临床,2014,30(6):184-186.
[12] JELEAZCOV C,IHMSEN H,SAARI T I,et al.Patientcontrolled analgesia with target-controlled infusion of hydromorphone in postoperative pain therapy[J].Anesthesiology,2016,124(1):56-68.
[13] CURTIS M J,WALKER M J.Quantification of arrhythmias using scoring systems:an examination of seven scores in an in vivo model of regional myocardial ischaemia[J].Cardiovasc Res,1988,22(9):656-665.
[14] DE HERT S,MOERMAN A.Myocardial injury and protection related to cardiopulmonary bypass[J]. Best Practice Res Clin Anaesth,2015,29(2):137-149.
[15] DE JONG J S S G,MARSMAN R F,HENRIQUES J P S,et al. Prognosis among survivors of primary ventricular fibrillation in the percutaneous coronary intervention era[J].Am Heart J,2009,158(3):467-472.
[16] OSMANCIK P P,STROS P,HERMAN D.Inhospital arrhythmias in patients with acute myocardial infarction-the relation to the reperfusion strategy and their prognostic impact[J].Acute Card Care,2008,10(1):15-25.
[17] KANNO S,SAFFITZ J E.The role of myocardial gap junctions in electrical conduction and arrhythmogenesis[J].Cardiov Pathol,2001,10(4):169-177.
[18] MIURA T,YANO T,NAITOH K,et al.Opioid receptor activation before ischemia reduces gap junction permeability in ischemic myocardium by PKC-mediated phosphorylation of connexin 43[J].AJP:Heart Circul Physiol,2007,293(3):H1425-H1431.
[19] TU I,YEN H D,CHENG H,et al.Baicalein protects chicken embryonic cardiomyocyte against hypoxia-reoxygenation injury viaμ-andδ-but notκ-opioid receptor signaling[J].Eur J Pharm,2008,588(2/3):251-258.