激动M3-mAchR对乌头碱诱发大鼠心律失常的保护机制的研究
|
摘要
目的通过乌头碱诱导的大鼠心律失常模型,研究匹罗卡品对心律失常的可能保护机制。方法 32只Wistar大鼠,雌雄不限,随机分成4组:模型组、4-二甲基氨基吡啶(4-DAMP)组、匹罗卡品组、匹罗卡品+4-DAMP组。4-DAMP组和匹罗卡品组大鼠,在股静脉静注乌头碱之前,分别提前5 min静注4-DAMP(0.12μg/kg)和匹罗卡品(0.2 mg/kg)。匹罗卡品+4-DAMP组大鼠,先给与M3-mAChR拮抗剂4-DAMP,5 min之后再给与匹罗卡品,再5min之后,用灌流泵经大鼠股静脉灌输乌头碱(0.06μg/min)诱发心律失常。观察每组大鼠的心律失常的严重程度,直至实验大鼠死亡。比较四组大鼠心律失常发生的起始时间、心肌室速和室颤发生时程、Mest评分和存活时间。采用激光共聚焦显微镜观察分离的心肌细胞内钙浓度变化。结果与模型组比较,匹罗卡品显著延迟大鼠心律失常发生的起始时间(P<0.001),减少大鼠心肌室速发生时程(P<0.001)和室颤的发生时程(P<0.001),降低心律失常的严重性和Mest评分(P<0.001),增加大鼠的存活时间(P<0.001)。预先给与匹罗卡品能够显著降低乌头碱诱导的心肌细胞发生的钙浓度的增加(P<0.05),与匹罗卡品组比较M3-mAchR选择性拮抗剂4-DAMP能部分阻断匹罗卡品对心律失常的保护作用(P<0.001),但4-DAMP本身对乌头碱诱导的心律失常并没有保护作用(P>0.05)。结论匹罗卡品对乌头碱诱导的大鼠心律失常具有保护作用,这种作用可能是通过激活M3-mAChR产生的,其机制可能与Ca~(2+)有关。
Objective To explore the underlying protective mechanism of pilocarpine by using aconitine-induced rat arrhythmia models.Methods Total 32 rats of either sex were randomly divided into 4 groups: model group,4-DAMP,pilocarpine,pilocarpine + 4-DAMP. Pilocarpine( 0. 2 mg/kg,intravenous) or 4-DAMP( 0. 12 mg/kg,intravenous) were administrated 5 minutes before aconitine respectively. For pilocarpine + 4-DAMP group,4-DAMP( 0. 12 mg/kg,intravenous),M3-mAChR antagonist was administrated 5 minutes before pilocarpine to verify the effect of M3-mAChR,then aconitine was injected via the femoral vein catheter with micro-infusion pump at a velocity of 0. 06 mg/min to induce ventricular arrhythmias. In each group,the arrhythmic severity was observed until the death of tested rats. Confocal microscopy was used to measure intracellular free-calcium concentrations( [Ca~(2+)]i) in isolated myocytes. Results Compared with the model group,pilocarpine significantly delayed onset of arrhythmias( P < 0. 001),decreased the time course of ventricular tachycardia( P < 0. 001) and fibrillation( P< 0. 001),reduced arrhythmia score( P < 0. 001),and increased the survival time of arrhythmic rats( P < 0. 001). [Ca~(2+)]i overload induced by aconitine was reduced in isolated myocytes pretreated with pilocarpine( P < 0. 05). Compared with the pilocarpine group,M3-muscarinic acetylcholine receptor( mAChR) antagonist 4-DAMP partially abolished the beneficial effects of pilocarpine( P < 0. 001),while 4-DAMP played no protection role in aconitine-induced rat arrhythmia. Conclusion Pilocarpine produced antiarrhythmic actions on arrhythmic rat induced by aconitine via stimulating the cardiac M3-mAChR. The mechanism may be related to the improvement of Ca~(2+) handling.
Objective To explore the underlying protective mechanism of pilocarpine by using aconitine-induced rat arrhythmia models.Methods Total 32 rats of either sex were randomly divided into 4 groups: model group,4-DAMP,pilocarpine,pilocarpine + 4-DAMP. Pilocarpine( 0. 2 mg/kg,intravenous) or 4-DAMP( 0. 12 mg/kg,intravenous) were administrated 5 minutes before aconitine respectively. For pilocarpine + 4-DAMP group,4-DAMP( 0. 12 mg/kg,intravenous),M3-mAChR antagonist was administrated 5 minutes before pilocarpine to verify the effect of M3-mAChR,then aconitine was injected via the femoral vein catheter with micro-infusion pump at a velocity of 0. 06 mg/min to induce ventricular arrhythmias. In each group,the arrhythmic severity was observed until the death of tested rats. Confocal microscopy was used to measure intracellular free-calcium concentrations( [Ca~(2+)]i) in isolated myocytes. Results Compared with the model group,pilocarpine significantly delayed onset of arrhythmias( P < 0. 001),decreased the time course of ventricular tachycardia( P < 0. 001) and fibrillation( P< 0. 001),reduced arrhythmia score( P < 0. 001),and increased the survival time of arrhythmic rats( P < 0. 001). [Ca~(2+)]i overload induced by aconitine was reduced in isolated myocytes pretreated with pilocarpine( P < 0. 05). Compared with the pilocarpine group,M3-muscarinic acetylcholine receptor( mAChR) antagonist 4-DAMP partially abolished the beneficial effects of pilocarpine( P < 0. 001),while 4-DAMP played no protection role in aconitine-induced rat arrhythmia. Conclusion Pilocarpine produced antiarrhythmic actions on arrhythmic rat induced by aconitine via stimulating the cardiac M3-mAChR. The mechanism may be related to the improvement of Ca~(2+) handling.
引文
[1]Khurana S,Jadeja R,Twaddell W,et al.Effect of modulating M3 muscarinic receptor activity on azoxymethane-induced liver injury in mice[J].Biochem Pharmacol,2013,86(2):329-338.
[2]Ponicke K,Heinroth-Hoffmann I,Brodde OE.Demonstration of functional M3 muscarinic receptors in ventricular cardiomyocytes of adult rats[J].Br J Pharmacol,2003,138(1):156-160.
[3]Hua N,Wei X,Liu X,et al.A novel muscarinic antagonist R2HBJJ inhibits non-small cell lung cancer cell growth and arrests the cell cycle in G0/G1[J].PLo S One,2012,7(12):e53170.
[4]Liu Y,Wang S,Wang C,et al.Upregulation of M muscarinic receptor inhibits cardiac hypertrophy induced by angiotensin II[J].J Transl Med,2013,11:209.
[5]Harvey RD.Muscarinic receptor agonists and antagonists:effects on cardiovascular function[J].Handb Exp Pharmacol,2012,(208):299-316.
[6]Abramochkin DV,Tapilina SV,Sukhova GS.Effect of selective stimulation of muscarinic M3 cholinoceptors on electrical and contractile activity of rat ventricular myocardium[J].Bull Exp Biol Med,2013,154(3):295-298.
[7]Wang S,Han HM,Jiang YN.Activation of cardiac M3 muscarinic acetylcholine receptors has cardioprotective effects against ischaemia-induced arrhythmia[J].Clin Exp Pharmacol Physiol,2012,39(4):343-349.
[8]Hang P,Zhao J,Qi J.Novel insight into the pervasive role of M3 muscarinic receptor in cardiac disease[J].Curr Drug Targets,2013,14(3):372-377.
[9]Li DL,Liu Y,Wang L,et al.Protective effects of pilocarpine on myocardial ischemia-reperfusion injury[J].Chin Pharm J,2007,42(19):1473-1476.
[10]解春花.匹鲁卡品对Wistar大鼠心律失常的影响[J].中国临床药理学杂志,2012,28(7):516-517,521.
[11]胡一冰,彭成.一种大鼠心律失常模型的建立[J].四川动物,2010,29(4):627-629.
[12]赵瑞富.防控钙通道相关室性心律失常的新靶点-钙库超载诱导钙释放(SOICR)[D].武汉:华中科技大学,2014.
[13]余丞浩.双酯型乌头碱致心肌细胞自发性钙释放机制研究[D].北京:中国中医科学院,2015.
[14]王相冲.乌头碱和新乌头碱致心律失常作用比较及其细胞学机制[D].石家庄:河北医科大学,2013.
[15]Gong DM,Shan HL,Zhou YH,et al.The ion targets of arrhythmias induced by ouabain and aconitine in guinea pig and rat ventricular myocytes[J].Yao Xue Xue Bao,2004,39(5):328-332.
[2]Ponicke K,Heinroth-Hoffmann I,Brodde OE.Demonstration of functional M3 muscarinic receptors in ventricular cardiomyocytes of adult rats[J].Br J Pharmacol,2003,138(1):156-160.
[3]Hua N,Wei X,Liu X,et al.A novel muscarinic antagonist R2HBJJ inhibits non-small cell lung cancer cell growth and arrests the cell cycle in G0/G1[J].PLo S One,2012,7(12):e53170.
[4]Liu Y,Wang S,Wang C,et al.Upregulation of M muscarinic receptor inhibits cardiac hypertrophy induced by angiotensin II[J].J Transl Med,2013,11:209.
[5]Harvey RD.Muscarinic receptor agonists and antagonists:effects on cardiovascular function[J].Handb Exp Pharmacol,2012,(208):299-316.
[6]Abramochkin DV,Tapilina SV,Sukhova GS.Effect of selective stimulation of muscarinic M3 cholinoceptors on electrical and contractile activity of rat ventricular myocardium[J].Bull Exp Biol Med,2013,154(3):295-298.
[7]Wang S,Han HM,Jiang YN.Activation of cardiac M3 muscarinic acetylcholine receptors has cardioprotective effects against ischaemia-induced arrhythmia[J].Clin Exp Pharmacol Physiol,2012,39(4):343-349.
[8]Hang P,Zhao J,Qi J.Novel insight into the pervasive role of M3 muscarinic receptor in cardiac disease[J].Curr Drug Targets,2013,14(3):372-377.
[9]Li DL,Liu Y,Wang L,et al.Protective effects of pilocarpine on myocardial ischemia-reperfusion injury[J].Chin Pharm J,2007,42(19):1473-1476.
[10]解春花.匹鲁卡品对Wistar大鼠心律失常的影响[J].中国临床药理学杂志,2012,28(7):516-517,521.
[11]胡一冰,彭成.一种大鼠心律失常模型的建立[J].四川动物,2010,29(4):627-629.
[12]赵瑞富.防控钙通道相关室性心律失常的新靶点-钙库超载诱导钙释放(SOICR)[D].武汉:华中科技大学,2014.
[13]余丞浩.双酯型乌头碱致心肌细胞自发性钙释放机制研究[D].北京:中国中医科学院,2015.
[14]王相冲.乌头碱和新乌头碱致心律失常作用比较及其细胞学机制[D].石家庄:河北医科大学,2013.
[15]Gong DM,Shan HL,Zhou YH,et al.The ion targets of arrhythmias induced by ouabain and aconitine in guinea pig and rat ventricular myocytes[J].Yao Xue Xue Bao,2004,39(5):328-332.