薯蓣皂苷对大鼠心肌收缩与钠-钙交换体的影响
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摘要
目的:观察薯蓣皂苷(Dio)对大鼠心肌收缩作用以及胞内Ca~(2+)浓度的影响,并初步探讨其作用机制与Na~+-Ca~(2+)交换体(NCX)的关系。方法:采用Langendorff逆行主动脉灌流法对大鼠离体心脏进行灌流,利用压力感受器插管法测定左心室相关心功能参数,记录及其在应用NCX选择性抑制剂SEA0400情况下对左心室收缩压(LVSP)、左心室舒张末期压(LVEDP)、左心室内压最大上升/下降速率(±dp/dt_(max))以及心率(HR)的影响;利用激光共聚焦显微观察薯蓣皂苷及SEA0400对大鼠心肌细胞H9c2细胞内Ca~(2+)浓度的影响。结果:离体心脏灌流结果显示,1μmol/L Dio可显著增加LVSP,增加约19.7%(P<0.01);增加左室内压最大上升速率(+dp/dt_(max)),增加约9.6%;激光共聚焦测定Ca~(2+)荧光强度实验结果显示:1μmol/L Dio可使H9c2细胞中Ca~(2+)相对荧光强度增加(P<0.01);而在SEA0400存在的情况下,1μmol/L的Dio使细胞内Ca~(2+)相对荧光强度变为(17.09±0.63),给予Dio后差异有显著性(P<0.01)。在细胞液中无Ca~(2+)或无Na~+时,给予1μmol/L的Dio使Ca~(2+)相对荧光强度减小,与给予1μmol/L的Dio差异有显著性(P<0.01)。结论:Dio可增加左心室收缩压和最大上升速率,表现正性肌力作用;Dio可使细胞内Ca~(2+)浓度增加,其作用机制与增加Na~+内流,促进NCX反向转运有关。
Objective: To investigate the inotropic effects of dioscin(Dio)in rat isolated-heart and intracellular free calcium concentration in isolated rat ventricular myocytes and to explore its mechanism preliminarily. Methods: Left ventricle contractile function was measured using the Langendorff non-recirculating mode of isolated rat heart perfusion. Effects of dioscin and dioscin + SEA0400,sodium-calcium exchanger( NCX) inhibitor,were investigated by measuring left ventricular systolic pressure( LVSP) and left ventricular end diastolic pressure( LVEDP). Also,heart rate( HR),peak rate of rise / fall of left ventricular pressure( ± dp / dt_(max)) of isolated rat heart were calculated; Effects of dioscin and SEA0400 on intracellular free calcium concentration in rat H9c2 cells were measured by Fluo3-AM and then detected the fluorescence intensity with confocal microscopy. Results: With 1 μmol / L dioscin,LVSP was significantly increased by 19. 7%( P < 0. 01) and dp / dt_(max)was increased by 9. 6%; With 1 μmol / L dioscin,the relative fluorescence intensity of intracellular free calcium concentrations were strong significantly( P < 0. 01). While in presence of SEA0400,the relative fluorescence intensity was changed to 17. 09 ± 0. 63( P < 0. 01) by 1 μmol / L dioscin. With 1 μmol / L dioscin,the relative fluorescence intensity was weak( P < 0. 01) without calcium or sodium in the extracellular fluid. Conclusion: Dioscin shows positive inotropic effect on isolated rat heart,enhancing the LVSP and + dp / dt_(max); Dioscin increases the intracellular concentration of Ca~(2+) in the cardiac myocytes by increasing Na~+influx and facilitating the reverse mode of the sodium-calcium exchanger.
Objective: To investigate the inotropic effects of dioscin(Dio)in rat isolated-heart and intracellular free calcium concentration in isolated rat ventricular myocytes and to explore its mechanism preliminarily. Methods: Left ventricle contractile function was measured using the Langendorff non-recirculating mode of isolated rat heart perfusion. Effects of dioscin and dioscin + SEA0400,sodium-calcium exchanger( NCX) inhibitor,were investigated by measuring left ventricular systolic pressure( LVSP) and left ventricular end diastolic pressure( LVEDP). Also,heart rate( HR),peak rate of rise / fall of left ventricular pressure( ± dp / dt_(max)) of isolated rat heart were calculated; Effects of dioscin and SEA0400 on intracellular free calcium concentration in rat H9c2 cells were measured by Fluo3-AM and then detected the fluorescence intensity with confocal microscopy. Results: With 1 μmol / L dioscin,LVSP was significantly increased by 19. 7%( P < 0. 01) and dp / dt_(max)was increased by 9. 6%; With 1 μmol / L dioscin,the relative fluorescence intensity of intracellular free calcium concentrations were strong significantly( P < 0. 01). While in presence of SEA0400,the relative fluorescence intensity was changed to 17. 09 ± 0. 63( P < 0. 01) by 1 μmol / L dioscin. With 1 μmol / L dioscin,the relative fluorescence intensity was weak( P < 0. 01) without calcium or sodium in the extracellular fluid. Conclusion: Dioscin shows positive inotropic effect on isolated rat heart,enhancing the LVSP and + dp / dt_(max); Dioscin increases the intracellular concentration of Ca~(2+) in the cardiac myocytes by increasing Na~+influx and facilitating the reverse mode of the sodium-calcium exchanger.
引文
[1]徐毅,罗卓卡,黄霏霏,等.正性肌力药物作用靶点的研究进展[J].中国新药杂志,2012,21(3):268-272.
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[11]Birinyi P,Acsai K,Bányász T,et al.Effects of SEA0400 and KB-R7943 on Na+/Ca2+exchange current and L-type Ca2+current in canine ventricular myocytes[J].Naunyn Schmiedebergs Arch Pharmacol,2005,372(1):63-70.
[12]Matsuda T,Arakawa N,Takuma K,et al.SEA0400,a novel and selective inhibitor of the Na+-Ca2+exchanger,attenuates reperfusion injury in the in vitro and in vivo cerebral ischemic models[J].J Pharmacol Exp Ther,2001,298(1):249-256.
[13]Iwamoto T,Kita S,Uehara A,et al.Molecular determinants of Na+/Ca2+exchange(NCX1)inhibition by SEA0400[J].J Bio Chem,2004,279(9):7544-7553.
[14]汪玲芳,赵云茜,高卫真,等.薯蓣皂苷增强大鼠缺血再灌注损伤后心肌抗氧化能力[J].中药药理与临床,2009,25(5):44-46.
[15]胡长鹰,于文喜.山药皂苷及其对离体心脏缺血再灌注损伤的保护作用[J].食品工业科技,2011,(2):309-312.
[16]Kaneko Y,Nakajima T,Irie T,et al.Ventricular fibrillation following bidirectional tachycardia due to digitalis toxicity[J].Intern Med,2011,50(19):2243-2243.
[17]刘静,吴红,丛恬马先,等.薯蓣皂苷含药血清对大鼠心室肌细胞钠离子通道的影响[J].中草药,2014,45(16):2370-2374.
[18]Farkas AS,Acsai K,Nagy N,et al.Na+/Ca2+exchanger inhibition exerts a positive inotropic effect in the rat heart,but fails to influence the contractility of the rabbit heart[J].Br J Pharmacol,2008,154(1):93-104.
[2]Pott C,Eckardt L,Goldhaber JI.Triple Threat:The Na+/Ca2+Exchanger in the Pathophysiology of Cardiac Arrhythmia,Ischemia and Heart Failure[J].Curr Drug Targets,2011,12(5):737-747.
[3]Passos AG,Gondim AN,Roman-Campos D,et al.The positive inotropic effect of the ethyl acetate fraction from Erythrinavelutina leaves on the mammalian myocardium:the role of adrenergic receptors[J].J Pharm Pharmacol,2013,65(6):928-936.
[4]Xie Z,Xie J.The Na/K-ATPase-mediated signal transduction as a target for new drug development[J].Front Biosci,2005,10:3100-3109.
[5]Chen L,Xu Y,Li W,et al.The novel compound liguzinediol exerts positive inotropic effects in isolated rat heart via sarcoplasmic reticulum Ca2+ATPase-dependent mechanism[J].Life Sci,2012,91(11):402-408.
[6]张伟锋,刘宝山.薯蓣皂苷的药理作用研究进展[J].世界中西医结合杂志,2010,5(6):543-545.
[7]侯娟,何文辉,王明霞,等.薯蓣皂苷的药理作用[J].河北医药,2004,26(1):71.
[8]王晓鹏,陆羡.薯蓣皂苷生物活性研究新进展[J].国外医学:中医中药分册,2004,26(3):138-141.
[9]宋宇,梁长青,何忠梅,等.薯蓣皂苷元体外抗肿瘤作用的研究[J].中国肿瘤,2004,13(10):651-653.
[10]Hobai IA,O'Rourke B.The potential of Na+/Ca2+exchange blockers in the treatment of cardiac disease[J].Expert Opin Inv Drugs,2004,13(6):653-664.
[11]Birinyi P,Acsai K,Bányász T,et al.Effects of SEA0400 and KB-R7943 on Na+/Ca2+exchange current and L-type Ca2+current in canine ventricular myocytes[J].Naunyn Schmiedebergs Arch Pharmacol,2005,372(1):63-70.
[12]Matsuda T,Arakawa N,Takuma K,et al.SEA0400,a novel and selective inhibitor of the Na+-Ca2+exchanger,attenuates reperfusion injury in the in vitro and in vivo cerebral ischemic models[J].J Pharmacol Exp Ther,2001,298(1):249-256.
[13]Iwamoto T,Kita S,Uehara A,et al.Molecular determinants of Na+/Ca2+exchange(NCX1)inhibition by SEA0400[J].J Bio Chem,2004,279(9):7544-7553.
[14]汪玲芳,赵云茜,高卫真,等.薯蓣皂苷增强大鼠缺血再灌注损伤后心肌抗氧化能力[J].中药药理与临床,2009,25(5):44-46.
[15]胡长鹰,于文喜.山药皂苷及其对离体心脏缺血再灌注损伤的保护作用[J].食品工业科技,2011,(2):309-312.
[16]Kaneko Y,Nakajima T,Irie T,et al.Ventricular fibrillation following bidirectional tachycardia due to digitalis toxicity[J].Intern Med,2011,50(19):2243-2243.
[17]刘静,吴红,丛恬马先,等.薯蓣皂苷含药血清对大鼠心室肌细胞钠离子通道的影响[J].中草药,2014,45(16):2370-2374.
[18]Farkas AS,Acsai K,Nagy N,et al.Na+/Ca2+exchanger inhibition exerts a positive inotropic effect in the rat heart,but fails to influence the contractility of the rabbit heart[J].Br J Pharmacol,2008,154(1):93-104.