乙酰胆碱对离体豚鼠心房肌动作电位及其脱敏的影响

目的 :研究迷走神经递质乙酰胆碱 （ACh）对离体豚鼠心房肌细胞动作电位 （AP）的影响及其对心房肌的脱敏 ,探讨脱敏可能发生的机制。方法 :采用标准玻璃微电极细胞内记录方法 ,观察不同浓度 （0 .0 1,0 .1,1μm ol/ L） ACh对心房肌细胞 AP的影响及对动作电位时程 （APD）和收缩力的脱敏现象 ,并观察了受体或通道水平的阻断剂阿托品、氯化铯 （Cs Cl）对 ACh所致的心房肌 APD脱敏的影响。结果 :10 .0 1,0 .1,1μm ol/ L ACh分别缩短 APD的变化率为 （6 .0 2± 1.0 4） % ,（14.2 0± 3.79） % ,（30 .2 0± 3.6 5 ） %。 2 0 .0 1μmol/ L ACh对心房肌细胞没有脱敏 ;0 .1μm ol/ L ACh对心房肌细胞 APD有轻微脱敏 ,脱敏持续时间为 1m in;而 1μmol/ L ACh对心房肌细胞 APD脱敏明显 ,脱敏持续时间为 5 m in。 31μmol/ L 阿托品与 2 0 m mol/ L Cs Cl并不能阻断 1μmol/ L ACh对心房肌细胞APD的脱敏。结论 :ACh缩短 APD的作用随浓度增大而增大 ;ACh对离体豚鼠心房肌细胞 APD的脱敏有浓度依赖性与时间依赖性 ;脱敏的发生可能与毒蕈碱型胆碱能受体及 Ik,ACh电流有关。     

溶血磷脂酸对豚鼠心室肌动作电位及延迟整流钾电流的影响

目的观察溶血磷脂酸（LPA）对离体豚鼠心室乳头肌动作电位及心室肌细胞延迟整流钾电流的影响。方法采用标准玻璃微电极技术记录豚鼠乳头肌动作电位。应用全细胞电压钳方法记录心室肌细胞延迟整流钾电流（Ik）。结果LPA0．1、1．0、10umol/L可浓度依赖性增加心室肌动作电位幅度（APA）（P〈0．05，P〈0．01，P〈0．01），延长动作电位50％、90％时程（APD50、APD90）（P〈0．05），钾通道阻断剂TEA可部分阻断LPA对APD50的延长作用。LPA0．1、1．0、10umol／L可明显抑制Ik（P〈0．05）。结论LPA可增加豚鼠心脏乳头肌动作电位幅值、延长动作电位时程，并抑制豚鼠心室肌细胞Ik。     

乙酰胆碱诱发的豚鼠心室肌反跳作用研究

观察乙酰胆碱 (ACh)诱发的豚鼠心室肌肌力及钙电流的反跳作用 ,并探讨其作用机制。采用豚鼠离体左室乳头肌观察 1μmol/LACh单独对心室肌收缩力 (Fc)的影响及在 10nmol/L异丙肾上腺素 (Iso)存在时对Fc的影响 ,并应用膜片钳全细胞记录方法分别观察 1μmol/LACh和 1μmol/LACh +10nmol/LIso及迅速洗脱ACh对豚鼠心室肌细胞L型钙通道电流 (ICa L)的影响。结果 :1μmol/LACh对心室肌Fc有直接抑制作用 ,抑制率为 30 .8%± 10 .7%(P <0 .0 5 ) ,而快速洗脱ACh后 ,Fc反跳性增强了 2 5 .5 %± 10 .3% (n =7,P <0 .0 1)。而在应用 10nmol/LIso后 1μmol/LACh对Fc有间接的抑制作用 ,快速冲洗ACh后亦引起Fc的反跳性增强 ,与Iso组相比增强了 2 7.1%±13.2 % (n =7,P <0 .0 1)。 1μmol/LACh对心室肌细胞基础峰电流无明显影响 (n =8,P >0 .0 5 ) ;而以基础ICa L峰值(931± 16 1pA)作对照 ,在加入 10nmol/LIso后 ,ICa L峰电流增强到 1889± 331pA(n =7,P <0 .0 1) ;再给予 10nmol/LIso +1μmol/LACh快速灌流 2min ,峰电流降低为 15 12± 2 0 2pA(P <0 .0 1) ,用含 10nmol/LIso的细胞外液快速洗脱ACh ,峰电流增强到 2 10 7± 2 0 5pA ,较Iso组反跳性增强了 15 .8%± 4 .0 % (n =7,P <0 .0 1)。结论 :ACh对豚鼠心室肌收缩     

不同浓度的乙醇对离体豚鼠心肌动作电位的影响

目的 观察不同浓度的乙醇对豚鼠心肌电生理特性的影响。方法 采用标准玻璃微电极细胞内记录技术,记录离体豚鼠心肌细胞的动作电位。观察12.5,25.0,50.0,100.0及200.0 mmol/L乙醇对心房肌和心室乳头肌动作电位各参数的改变。结果 12.5~100.0 mmol/L 乙醇对心室乳头肌细胞的细胞膜静息电位（RMP）、动作电位幅值（APA）、动作电位时程（APD）、AP复极至50%的时程（APD50）及AP复极至90%的时程（APD90）等参数无明显影响;而200.0 mmol/L乙醇可显著降低APA（P<0.05）,50.0~200.0 mmol/L乙醇可明显延长心房肌细胞APD,APD50和APD90（P<0.05或P<0.01）。结论 乙醇对离体豚鼠心房肌、心室乳头肌电生理特性的影响有着明显差异。心房肌APD的延长可能与心房肌中分布的特殊离子通道被阻滞有关。     

阿米洛利对心肌复极的影响及离子机制研究

目的 :研究阿米洛利 （Amiloride）对心肌复极的影响及离子机制。方法 :用标准微电极技术记录阿米洛利对豚鼠乳头状肌细胞动作电位的影响 ;用膜片钳技术记录电压依赖性的钾电流。结果 :10 μmol/L、10 0 μmol/LAmiloride均可降低豚鼠乳头状肌动作电位幅度 ,延长动作电位时程 ;当累积浓度达 10 0 μmol/L时 ,Amiloride轻微抑制快激活延迟整流钾电流 （Ikr） ,对慢激活延迟整流钾电流 （Iks）无影响。 （1～ 10 0 ） μmol/L的Amiloride浓度依赖性地抑制内向整流钾电流 （Ik1）。结论 :阿米洛利通过抑制电压依赖性的钾电流 ,延长复极以发挥抗心律失常的作用。     

艾司洛尔对心室肌细胞动作电位及L-型钙离子通道的影响

目的 观察艾司洛尔对豚鼠心室肌细胞动作电位(AP)和L-型钙离子通道的影响.方法 Langendorff离体心脏逆向灌流法分离豚鼠心室肌细胞,随机选取心室肌细胞分为正常对照组和艾司洛尔(50 μmol/L和100 μmol/L)组.应用全细胞电流钳模式记录心室肌细胞AP,应用电压钳模式记录L-型钙离子通道电流(ICa-L).结果 艾司洛尔(100 μmol/L )可使心肌细胞AP时程APD20、APD50明显缩短(P<0.05),心室肌细胞ICa-L峰值电流明显降低(P<0.05).结论 艾司洛尔缩短心室肌细胞AP时程和抑制钙通道可能是其抑制交感风暴的机制之一.     

乙酰胆碱对豚鼠心房、心室直接作用及其机制的比较研究

目的:自主神经系统是哺乳动物心脏的重要调节因素之一。一般认为,交感神经支配心脏的各个部位,而副交感神经则主要支配室上性组织。副交感神经递质乙酰胆碱(ACh)激活突触后膜上的毒蕈碱型胆碱受体(M- Ch R) ,对心房和心室产生不同的作用。ACh主要通过毒蕈碱敏感性钾通道(KACh)对心房产生直接负性作用,对心室则在β-肾上腺素受体被预先激动后产生间接的抑制作用。ACh对哺乳动物的心室肌无直接负性变力作用,雪貂、大鼠例外。对于豚鼠的心室肌来说,资料显示结果不一。而且ACh对心房、心室作用的比较以及收缩力与动作电位时程(APD)的关系…     

卡维地洛对豚鼠心室肌细胞的电生理作用

目的:观察卡维地洛对豚鼠心室肌细胞膜离子流及跨膜动作电位的影响,探讨卡维地洛对心室肌细胞的直接电生理作用.方法:①应用膜片钳全细胞记录技术记录单个豚鼠心室肌细胞膜L型钙内流(ICa-L)、内向整流性钾流(IJ1)、延迟整流性钾流(IK)和快钠内流(INa),观察不同浓度的卡维地洛对各离子流的影响.②用标觋准微电极技术记录豚鼠右室乳头肌细胞动作电位,观察不同浓度卡维地洛对动作电位各参数的影响.结果:①卡维地洛浓度依赖性地抑制ICa-L、INa,半数抑制浓度(IC50)分别为3.18,0.16 μmol/L.②卡维地洛25μmol/L对IK1最大内向和外向电流差异无统计学意义(n=5,P＞0.05),但使其整流范围由-20～ 40 mV增大至-20～ 80 mV.③卡维地洛0.3μmol/L以上即可浓度依赖性抑制IK尾电流(IK.tail),且对-10 mV钳制电压下IK.tail的抑制作用明显大于对 50 mV下的抑制,在高浓度还抑制缓慢激活型IK(IKs),IC50为12.43μmol/L.④卡维地洛呈反转频率依赖性延长动作电位时程(APD90),还降低动作电位幅度(APA)和0相最大去极化速率(Vmax).结论:卡维地洛低浓度下即可抑制心肌细胞INa、快速激活相IK,高浓度下抑制ICa-L、IKs,还降低Vmax、APA,呈反转频率依赖性延长APD90.     

别隐品碱抗心律失常效应及其对动作电位的影响

目的研究别隐品碱(ALL)抗心律失常效应及其对心室肌动作电位的影响.方法选择大鼠和豚鼠以静脉给乌头碱、哇巴因和氯化钡造动物心律失常的模型,观察ALL对心律失常的效应,分离豚鼠乳头状肌观察ALL对动作电位的影响.结果ALL(30和100mg/kg)对乌头碱、哇巴因和氯化钡致动物实验性心律失常有良好的对抗作用,使各种试剂诱发的室早、室速、室颤及停搏的剂量增加.ALL可降低动作电位上升幅度(APA)和0相最大上升速率(Vmax).3.0～100.0 μmol/L的ALL以浓度依赖性方式使离体乳头状肌动作单位时程(APD)延长,其中30.0μmol/L的ALL使APD90延长67.5%.结论ALL具有抗实验性心律失常的效应,此效应可能与其影响心肌组织的电生理密切相关.     

5-HT4 受体部分激动剂RS67506可作为心肌IK1通道抑制剂

目的 验证5-HT4受体部分激动剂RS67506对心室肌内向整流钾通道（IK1）和动作电位（action potential，AP）的作用特征。方法 取成年雄性SD大鼠心脏经Langendorff离体灌流、胶原酶法急性分离单个左心室肌细胞，采用膜片钳技术检测1~30 μmol/L RS67506对心室肌静息电位（resting potential，RP）、动作电位（action potential，AP ）及IK1通道电流的影响。结果 1、10、30 μmol/L RS67506可剂量依赖性降低静息电位（由-75.2±0.4 mV 分别降低至-72.7±0.5 mV，-70.1±0.4 mV和-67.9±0.9 mV）（P<0.01），延长动作电位复极时间（APD）（APD90由31.1±1.1ms 分别延长至34.7±0.6 ms，40.4±1.2ms和45.0±0.5ms）（P<0.05 或P<0.01）；同时抑制IK1，30 μmol/L RS67506可使IK1外向电流密度（-60 mV）降低58.4%（P<0.05）；内向电流密度（-100 mV）降低53.2%（P<0.01）。结论 RS67506对大鼠心室肌IK1和AP的作用特征符合IK1抑制剂的特点，可能作为大鼠心肌IK1抑制剂，为临床防治心律失常提供新的药理学工具药。     

Potentiating Effect of Acetylcholine on Stimulation by Isoproterenol of L-Type Ca2+ Current and Arrhythmogenic Triggered Activity in Guinea Pig Ventricular Myocytes

ACh Facilitation of Triggered Activity. Introduction : The objective of this study was to determine whether the effect of isoproterenol (Iso) to increase L-type Ca²⁺ current [I_ca(L)] and action potential duration (APD) was potentiated in ventricular myocytes following termination of an exposure of these cells to acetylcholine (ACh), and whether this potentiating effect of Ach could he arrhythmogenic.
Methods and Results : Transmembrane currents and potentials of guinea pig Isolated ventricular myocytes were measured using the whole cell, patch clamp technique. Stimulation of I_ca(L) and prolongation of APD caused by Iso (10 nmol/L) were attenuated in the presence of Ach (10 μ mol/L), but were transiently enhanced by 111%± 20% and 214%± 44%, respectively, following termination of a 2- to 4-minute exposure of myocytes to ACh. No changes were observed in the absence of Iso. Both the amplitude and incidence of isoinduced transient inward current, afterdepolarizations, and sustained triggered activity were greater immediately after termination of exposure to ACh than before application of ACh.
Conclusion : Stimulation by Iso of I_ca(L) is transiently enhanced in guinea pig ventricular myocytes following termination of exposure of these cells to ACh. The rebound increase of Iso-stimulated I_ca(L) is associated with an increase of APD and induction of arrhythmogenic triggered activity.     

丹参素对豚鼠心室肌细胞L-型钙通道的影响

目的　研究丹参素对豚鼠心室肌细胞膜L 型钙通道的影响 ,探讨丹参素在离子通道水平的药理机制。方法　急性酶解法获得豚鼠的单个心肌细胞 ,用标准的全细胞膜片钳技术记录钙电流。结果　在保持电位 (Holdingpoten tial,HP)为 - 80mV ,预先去极化至 - 4 0mV ,再去极化至0mV ,波宽为 30 0mS的刺激参数下 ,可记录到一具有电压和时间依赖性内向的电流 ,当用含 1μmol/L硝苯地平的台氏液灌流时该电流被迅速消除 ,在灌流液中加入丹参素 1mg/mL和 10mg/mL ,1mg/mL组对钙电流无明显改变 (P >0 0 5 ,n =5 ) ,10mg/mL组使钙电流减少 [- (7 76± 0 85 )pA/pFvs - (2 6 2 6± 0 5 9)pA/pFP <0 0 5 ,n =8],并使心肌细胞钙电流 -电压曲线上移 ,原有的电流 -电压依赖特征不变。结论　丹参素浓度依赖性地抑制心肌L 型钙通道     

氧化苦参碱对豚鼠心室肌细胞膜L-型钙通道的影响

研究氧化苦参碱 (Oxy)对豚鼠心室肌细胞膜L 型钙通道的影响 ,探讨Oxy在离子通道水平的药理作用机制。用急性酶解法分离豚鼠心室肌细胞 ,应用膜片钳全细胞记录技术 ,观察不同浓度的Oxy对L 型钙通道的影响。结果 :用 0 .0 1 ,0 .1 ,1 ,1 0 μmol/L可浓度依赖性地增加L 型钙电流 (ICa L)。在 0 .1 μmol/L时 ,给药后电流密度增加约 31 %(P <0 .0 1 ) ,可使心肌细胞钙电流 电压曲线下移 ,但激活电位、峰电位及反转电位无改变 ;Oxy使激活曲线向负电位方向变化 ,半数激活电压增加 ;对失活曲线无明显影响 ,未改变ICa L失活特性。结论 :Oxy可浓度依赖性和电压依赖性地增加心肌细胞膜L 型钙通道电流     

Antimalarial drugs inhibit the acetylcholine-receptor-operated potassium current in atrial myocytes

BACKGROUND: It has been reported that halofantrine, an antimalarial drug, was associated with electrocardiographic prolongation of the QT interval and ventricular arrhythmias. Inhibition of the delayed rectifier potassium channel, a voltage-gated potassium channel, by halofantrine was the likely underlying cellular mechanism for this cardiotoxicity. However, influences of anti-malarial drugs on the ligand-gated potassium channels have not been well-documented. The influences of three different antimalarial drugs, chloroquine, primaquine and pyrimethamine, on the acetylcholine-receptor-operated potassium current (I(K.ACh)), a ligand-gated potassium current, were compared with the effect of quinidine in isolated guinea pig atrial myocytes using patch-clamp techniques. METHODS: The whole-cell patch-clamp method was used in the present studies he I(K.ACh) was induced by extracellular application of carbachol (1 micromol/L) or intracellular loading of guanosine 5'-O-(3-thiotriphosphate) GTPgammaS (100 micromol/L) in acutely isolated guinea pig atrial myocytes. RESULTS: The I(K.ACh) induced by carbachol was inhibited by chloroquine, primaquine, pyrimethamine and quinidine in a concentration-dependent manner, and the concentrations required to produce 50% of the maximal inhibitory effect (IC(50) values) were 0.7, 2.5, 12 and 1.8 micromol/L, respectively. These drugs also inhibited the intracellular GTPgammaS-activated I(K.ACh), and the IC(50) values were 0.8,13,19 and 21 micromol/L, respectively. CONCLUSIONS: Chloroquine and pyrimethamine may inhibit I(K.ACh) by interacting with the muscarinic potassium channel itself and/or associated guanosine 5'-triphosphate-binding proteins, whereas primaquine and quinidine may mainly inhibit the current by the blockade of the muscarinic receptors. These results indicate that antimalarial drugs exert anticholinergic effects via different molecular mechanisms.     

17β-雌二醇对豚鼠心室肌细胞动作电位及L型钙通道电流的影响

探讨雌激素对心血管保护作用的机制,采用标准玻璃微电极技术及全细胞膜片钳技术观察17β-雌二醇（ES）对豚鼠心室乳头肌细胞动作电价的影响及其对分离的豚鼠心室肌细胞的L型钙通道电流的作用。结果：30μmol／LES灌流心肌30min可明显缩短动作电位时程;复极20％动作电位时程及复极50％动作电位时程分别由146±22ms,213±21ms缩短至87±6ms,107±11ms（P均＜0．01）。1,10,20,30μmol／LES是浓度依赖性抑制L型钙通道电流（P均＜0．01）,抑制率分别为8％,18％,33％,43％。结论：ES是一种内源性的钙离子通道拮抗剂,这可能是其心血管保护作用的重要机制之一．     

Sympathetic activation, ventricular repolarization and Ikr blockade: Implications for the antifibrillatory efficacy of potassium channel blocking agents

Objectives. The aim of the present study was to test, in vivo and in vitro, the influence of adrenergic activation on action potential prolongation induced by the potassium channel blocking agent d-sotalol.

Background. d-Sotalol is not effective against myocardial ischemia-dependent ventricular fibrillation in the presence of elevated sympathetic activity. Most potassium channel blockers, such as d-sotalol, affect only one of the two components of I_k (I_kr) but not the other (I_ks). I_ks is activated by isoproterenol. An unopposed activation of I_ks might account for the loss of anti-fibrillatory effect by d-sotalol in conditions of high sympathetic activity.

Methods. In nine anesthetized dogs we tested at constant heart rate (160 to 220 beats/min) the influences of left stellate ganglion stimulation on the monophasic action potential prolongation induced by d-sotalol. In two groups of isolated guinea pig ventricular myocytes we tested the effect of isoproterenol (10⁻⁹ mol/liter) on the action potential duration at five pacing rates (from 0.5 to 2., Hz) in the absence (n = 6) and in the presence (n = 8) of d-sotalol.

Results. In control conditions, both in vivo and in vitro, adrenergic stimulation did not significantly change action potential duration. d-Sotalol prolonged both monophasic action potential duration in dogs and action potential duration of guinea pig ventricular myocytes by 19% to 24%. Adrenergic activation, either left stellate ganglion stimulation in vivo or isoproterenol in vitro, reduced by 40% to 60% the prolongation of action potential duration produced by d-sotalol.

Conclusions. Sympathetic activation counteracts the effects of potassium channel blockers on the duration of repolarization and may impair their primary antifibrillatory mechanism. An intriguing clinical implication is that potassium channel blockers may not offer effective protection from malignant ischemic arrhythmias that occur in a setting of elevated sympathetic activity.     

Acetylcholine, Ca2+ overload and oscillatory potentials in isolated ventricular myocytes.

The events caused by overdrive-induced calcium overload were studied in guinea pig isolated ventricular myocytes. Overdrive may induce oscillatory potentials (Vos) and a prolonged depolarization (Vex), as well as the underlying currents (Ios and Iex, respectively). Acetylcholine (ACh) reduced or abolished these events, an action which was blocked by atropine. Norepinephrine exaggerated the effects of overdrive, and ACh markedly antagonized such an enhancement. Caffeine at low concentrations increased both Ios and Iex whereas at high concentrations caffeine abolished Ios but increased Iex. Quinacrine abolished both events. Voltage clamp depolarizing steps abolished Ios (and did not reverse it). Thus, the effects of Ca overload are antagonized by ACh in the absence and presence of sympathetic enhancement. Also, the mechanism underlying Ios appears to involve an electrogenic Ca extrusion and not an increase in a nonspecific conductance.     

Effects of sildenafil on cardiac repolarization

OBJECTIVES: Sudden death has occasionally been reported in patients taking sildenafil. The objective of this study was to investigate the effect of sildenafil on cardiac repolarization. METHODS: We used conventional microelectrode recording technique in isolated guinea pig papillary muscles and canine Purkinje fibers, whole-cell patch clamp techniques in guinea pig ventricular myocytes, and in vivo ECG measurements in guinea pigs. RESULTS: Action potential duration at 90% repolarization (APD(90)) was not affected by sildenafil in the therapeutic ranges (< or =1 microM), but shortened by higher concentration (> or =10 microM) in both guinea pig papillary muscles and canine Purkinje fibers. D-Sotalol prolonged APD(90) in the same preparations with concentrations > or =1 microM in a reverse frequency-dependent manner. Co-administration of sildenafil (10 and 30 microM) abolished the APD-prolonging effects of D-sotalol (30 microM) and amiodarone (100 microM). Sildenafil, with concentrations up to 30 microM, had no significant effect on both the rapid (I(Kr)) and the slow (I(Ks)) components of the delayed rectifier potassium currents in guinea pig ventricular myocytes. Sildenafil dose-dependently blocked L-type Ca(2+) current (I(Ca,L)), but had no effect on persistent Na(+) current in guinea pig ventricular myocytes. ECG recordings in intact guinea pigs revealed significant shortening of QTc interval by sildenafil (10 and 30 mg/kg orally). The QT-prolonging effects by D,L-sotalol (50 mg/kg) and amiodarone (100 mg/kg) were abolished by sildenafil (30 mg/kg). CONCLUSIONS: Sildenafil does not prolong cardiac repolarization. Instead, in supra-therapeutic concentrations, it accelerates cardiac repolarization, presumably through its blocking effect on I(Ca,L).