Regional Gradation of L-Type Calcium Currents in the Feline Heart with a Healed Myocardial Infarct

I_Ca in Healed Myocardial Infarction. Introduction: Abnormal action potentials in myocytes adjacent to > 2-month-uld feline LV myocardial infarcts (MI) may reflect alterations in Ca²⁺ currents (I_Ca). Methods and Results: We compared I_Ca, at 36°C, in subendocardial myocytes isolated from areas adjacent to MI and to I_Ca in cells from remote areas (> 4 mm away; REM) and control cells from similar regions in normal hearts. Control (CON) myocytes had membrane capacitance of 234 ± 10 pF (n = 81 cells) compared to 305 ± 14 pF in REM (71 cells; P < 0.05 from CON) and 237 ± 11 pF (n = 55 cells) in MI (not different from CON). From V_h=?40 mV, peak I_Ca elicited by test potentials (?35 to +70 mV) were significantly larger in CON (?1746 ± 123 pA) and REM (?1795 ± 142 pA) compared to Ml (?1352 ± 129 pA) (P < 0.05). Peak 1(11 density was significantly reduced in REM (?6.0 ± 0.4 pA/pF) or MI (?5.7 ± 0.4 pA/pF, P < 0.05) compared to CON (?7.5 ± 0.4 pA/pF). Double exponential I_Ca decay was similar among groups. Half-inactivation potential (V_0.5) was significantly shifted (hyperpolarizing direction) for MI (?29.1 ± 2.6 mV) and REM (?24.6 ± 1.2 mV) myocytes compared to ?20.3 ± 1.0 mV in CON. MI slope factor (k; 9.0 ± 0.5) was significantly different from CON (6.8 ± 0.3) and RKM (7.3 ± 0.4). No differences in time course of recovery from inactivation were noted. Five millimolar Ba²⁺₀ produced significant increases in I_Ca in CON and REM but an attenuated response in MI. Bay k8644 (1 μM) produced similar I_Ca increase in all groups. I_Ca increase due to isoproterenol (1 I_CaM) in MI and REM was half that in CON, but there were no differences in increased I_Ca responses among groups following phenylephrine (10 μM). Conclusion: Reduced I_Ca density in REM reflects cell hypertrophy, whereas altered I_Ca of MI may reflect altered channel structure and/or function.     

Ventricular Action Potential and L-Type Calcium Channel in Infarct-Induced Hypertrophy in Rats

Calcium Channel in Infarct-Induced Hypertrophied Rat Ventricle. Introduction : The present investigation was aimed at characterization of: (1) action potential parameters; and (2) L-type calcium channels in the hypertrophied ventricular tissue surviving an extensive healed myocardial infarction in the rat.
Methods and Results : Myocardial infarction was produced in Wistar rats by ligation of the left coronary artery. One to 2 months later, their hearts were subjected to electrophysiologic study. The main difference in subendocardial transmembrane potentials recorded with intracellular microelectrodes was an increase in action potential duration (APD). In the left ventricle, the infarcted/sham-operated APD ratio ranged from 2.7 to 7.2, whereas in the right ventricle it ranged from 1.6 to 2.3 in different regions. When compared with control ceils, ventricular myocytes from infarcted hearts were found to be larger (P < 0.01) and showed a reduction (P < 0.05) in L-type calcium current (I_Ca,L) density obtained by whole cell, patch clamp (at 0 mV: 4.44 ± 0.41 in infarcted vs 8.03 ± 1.22 pA/pF in normal). The time course of decay of the currents could be fitted by two exponential functions in both normal and infarcted hearts. There was a tendency toward an increase in the time constant of the slower component of inactivation, T₂, significant only at +20 mV (215 ± 25 vs 151 ± 15 msec).
Conclusions : Cardiac hypertrophy of healed infarction in rats is associated with lengthening of the action potential in both ventricles. The main alteration observed in I^Ca,Lwas a decrease in the current density. Thus, alteration of the calcium channel is not the determinant factor of APD increase     

兔急性心肌梗死后梗死周边带心肌细胞L-型钙通道的变化

探讨L型钙通道在急性心肌梗死 (AMI)后室性心律失常发生中的作用及其机制。方法 :以开胸冠状动脉结扎法制备兔AMI模型 ,1周后处死动物分离心室肌细胞 ,采用全细胞膜片钳记录技术观察梗死周边缺血带心外膜心室肌细胞L型钙通道电流 (ICa L)的变化 ,以正常心肌ICa L为对照。结果 :AMI 1周时兔梗死周边区心室肌细胞L型钙电流受到抑制 ,其电流峰值由正常状态下的 - 5 .58± 1 .53pA/pF(对照组 ,n =1 0 )降至 - 3 .52± 0 .93pA/pF(AMI组 ,n=6) ,最大峰电流下降 2 9.1 % ,P <0 .0 5 ,I V曲线上移 ;其失活曲线左移 ,半数最大失活电位由 - 1 3 .1± 4 .2mV左移至 - 2 5 .9± 7.0mV ,P <0 .0 5 ,失活速度加快。结论 :AMI后 1周梗死周边带心外膜心室肌细胞L型钙通道受抑制 ,可能为AMI后室性心律失常发生的机制之一。     

Effects of Simvastatin on Ion Channel Currents in Ventricular Myocytes from Acute Infarcted Heart of Normocholesterolemic Rabbits

Objectives To investigate the effects of simvastatin on membrane ionic currents in left ventricular myocytes of rabbit heart suffering from acute myocardial infarction （ AMI）, so as to explore the ionic mechanism of statin treatment for antiarrhythmia. Methods Forty-five New Zealand rabbits were randomly divided into three groups： AMI group, simvastatin intervention group （ Statin group） and sham-operated control group （CON）. Rabbits were infarcted by ligation of the left anterior descending coronary artery after administration of oral simvastatin 5 mg · kg^-1·d^-1 （Statin group） or placebo （AMI group） for 3 days. Single ventricular myocytes were isolated enzymatically from the epicardial zone of the infracted region 72 h later. Whole cell patch clamp technique was used to record membrane ionic currents, including sodium current （INa）, L-type calcium current （Ica-L） and transient outward potassium current （Ito）. Results （1） There was not significant difference in serum cholesterol concentration among three groups. （2） The peak INa current density （at -30 mV） was significantly decreased in AMI group （ -25.26±5.28, n = 13 ）, comparing with CON （ - 42. 78± 5.48, n = 16）, P 〈 0. 05, while it was significantly increased in Statin group （ - 39.83 ±5.65 pA/pF, n = 12） comparing with AMI group, P 〈0. 01 ; The peak Ica-L current density （ at 0 mV） was significantly decreased in AMI group （ -3. 43 ±0. 92 pA/pF, n = 13） comparing with CON （ -4. 56 ±1.01 pA/pF, n = 15）, P 〈0. 05, while it was significantly increased in Statin group （ -4. 18±0. 96 pA/pF, n = 12） comparing with AMI group, P 〈0. 05; The Ito current density （ at ＋ 60 mV） was significantly decreased in AMI group （ 11.41 ± 1.94 pA/pF, n = 13 ） comparing with CON （17.41 ±3.13 pA/pF, n = 15）, P 〈0. 01, while it was significantly increased in Statin group （16. 11 ± 2. 43 pA/pF, n = 14） comparing with AMI group, P 〈 0. 01. Conclusions AMI induces signific     

急性心肌梗死后一周对心室肌细胞瞬间外向钾电流的影响

研究急性心肌梗死 (AMI)心室肌细胞瞬间外向钾电流 (Ito)的变化。采用结扎兔冠状动脉左前降支的方法建立AMI动物模型 ,应用膜片钳全细胞记录方法 ,研究AMI后 1周心外膜梗死区心肌细胞Ito的变化。结果 :正常对照组 (n =16 )心肌细胞在 - 30mV激活 ,心肌梗死 (简称心梗 )组细胞在 - 2 0mV激活 ,均呈线性电压依赖性。心梗组梗死区细胞 (n =12 )Ito的电流密度明显下降 ,I V曲线明显下移。心梗组Ito电流密度 (去极化电位 +6 0mV时 )明显低于对照组 (7.4 7± 2 .39vs 17.39± 5 .2 4pA/pF ,P <0 .0 1)。结论 :AMI可引起心室肌细胞Ito电流密度下降 ,导致梗死区细胞动作电位平台期相对延长 ,复极异常 ,造成心肌细胞之间动作电位及不应期离散度增大 ,容易形成折返 ,此可能是导致心肌梗死后出现折返性室性心律失常的原因。     

Ghrelin对大鼠心室肌细胞L型钙电流的影响

目的：研究一种具有生长激素释放活性的脑肠肽（Ghrelin）对大鼠心室肌细胞L型钙电流的影响。方法：采用酶解消化法获得大鼠单个心室肌细胞,通过全细胞膜片钳技术研究不同浓度（10 nmol/L、100 nmol/L及1μmol/L） Ghrelin对L型钙电流的影响。结果：10 nmol/L、100 nmol/L及1μmol/L Ghrelin依次抑制大鼠心室肌细胞L型钙电流峰电位,抑制率分别为：8.95%±2.13%、31.18%±4.78%及64.63%±8.57%,（P<0.05）;I-V曲线上移,通道半数失活电压从（-1.34±1.9） mV分别降至（-8.04±1.32）mV、（9.76±1.17）mV及（-11.81±0.73）mV（P<0.05）,并延长失活后恢复时间由给药前τ值63.23±9.32,分别增至98.95±10.74、109.56±13.42和127.39±16.13,差异有统计学意义（P<0.05）。结论：Ghrelin通过加快L型钙电流的失活及延长失活后恢复时间,具有浓度依赖性地抑制L型钙电流。     

兔急性心肌梗死后二月心室肌细胞钠离子通道活性的变化

研究急性心肌梗死 (AMI)后心室肌细胞钠离子通道活性的变化。采用结扎兔冠状动脉左前降支的方法建立AMI动物模型 ,应用膜片钳全细胞记录方法 ,观察AMI后 2个月心外膜梗死区心肌细胞钠通道电流 (INa)的变化。结果 :①正常对照组INa电流密度峰值 (去极化电位 - 30mV时 )为 45 .5± 5 .33pA/pF(n =12 ) ,心肌梗死 (简称心梗 )组为 16 .4± 4.43pA/pF(n =13) ,心梗组较对照组明显下降 ,P <0 .0 1。心梗组INa电流 电压关系曲线较对照组明显下移。②心梗组INa失活曲线较对照组明显左移 (即向超级化方向移动 ) ,对照组半数失活电压 (V0 .5)为 - 76 .2± 5 .3mV(n =5 ) ,心梗组V0 .5为 - 82 .4± 5 .6mV(n =12 ) ,P <0 .0 5。③心梗组钠通道灭活后恢复时程较对照组减慢 ,恢复曲线下移。结论 :AMI可导致梗死区心室肌细胞INa下降、钠通道动力学发生变化 ,引起心肌传导速度下降和不应性延长 ,此可能是导致AMI后出现折返性室性心律失常的原因。     

Effects of Mibefradil, a T-Type Calcium Current Antagonist, on Electrophysiology of Purkinje Fibers that Survived in the Infarcted Canine Heart

INTRODUCTION: We studied the effects of mibefradil (MIB), a nondihydropyridine T-type Ca2+ channel antagonist, on T- and L-type Ca2+ (I(CaT), I(CaL)) currents in Purkinje myocytes dispersed from the subendocardium of the left ventricle of normal (NZPC) and 48-hour infarcted (IZPC) hearts. METHODS AND RESULTS: Currents were recorded with Cs+- and EGTA-rich pipettes and in Na+-K+-free external solutions to eliminate overlapping currents. In all cells, I(Ca) was reduced by MIB (0.1 to 10 microM). No change in the time course of decay of peak I(Ca) was noted. Average peak T/L ratio decreased in NZPCs but not IZPCs with 1 microM MIB. Steady-state availability of I(CaL) was altered with 1 microM MIB in both cell types (mean +/- SEM) (V0.5 = -22 +/- 4 mV for NZPC and -25 +/- 5 mV for IZPC before drug; -63 +/- 9 mV for NZPC and -67 +/- 6 mV for IZPC after drug; P < 0.05). For I(CaT), V0.5 (-50 +/- 3 mV for NZPC and -52 +/- 1 mV for IZPC before drug) shifted to -60 +/- 2 mV (NZPC) and -62 +/- 3 mV (IZPC) (P < 0.05) after drug. We also determined the effects of MIB on spontaneously beating Purkinje normal fibers and on depolarized abnormally automatic fibers from the infarcted heart using standard microelectrode techniques. When NZPC and IZPC fibers were superfused with [K+]o = 2.7 mM, MIB 3 microM and 10 microM had no effect on rate or the maximum diastolic potential, but action potential plateau shifted to more negative values, the slope of repolarization phase 3 decreased, and action potential duration increased. CONCLUSION: MIB blocks L- and T-type Ca2+ currents in Purkinje myocytes but lacks an effect on either normal or abnormal automaticity in Purkinje fibers.     

心肌梗死后大鼠左心室肌细胞钾通道的重构

目的:探讨心肌梗死后大鼠左心室肌细胞钾通道的重构及增加葡萄糖代谢对钾流的作用。方法:取体重200～250 g的雄性Sprague-Dawley(SD)大鼠8只,结扎左冠状动脉建立心肌梗死模型(心肌梗死组),采用酶解法获得单个左心室肌细胞,应用膜片钳全细胞记录技术记录钾电流。对照组5只不结扎冠状动脉。结果:心肌梗死组大鼠心脏重量、心脏／体重比及左心室肌细胞膜电容均较对照组显著增加,但瞬间外向性钾流(Ito)密度则显著降低,分别用1．5 mmol／L二氯乙酸及5 mmol/L丙酮酸在体外预处理心肌梗死后大鼠左心窒肌细胞4～5小时,瞬间外向性钾流密度恢复到对照组水平。结论:心肌梗死后大鼠左心室肌细胞钾通道存在重构,而增加葡萄糖代谢能使之恢复,提示糖代谢和钾通道功能间存在一定关系。     

Reduced Inward Rectifying and Increased E-4031-Sensitive K+ Current Density in Arrhythmogenic Subendocardial Purkinje Myocytes from the Infarcted Heart

Outward Currents in Purkinje Cells from 48-Hour Infarcted Heart. Introduction : Subendocardial Purkinje myocytes from the 4K-hour infarcted heart (IZPCs) have reduced resting potentials, possibly due to altered inwardly rectifying K⁺ currents I_KI. Abnormal depolarization-activated outward K⁺ currents could contribute to long triangularly shaped action potentials of IZPCs.
Methods and Results : We used whole cell patch recordings to compare cesium-sensitive I_KI and 4-aminopyridine (4-AP)-resistant, noninactivating sustained I_K between normal Purkinje myocytes (NZPCs) and IZPCs. IZPCs showed decreased net membrane currents. Two IZPC groups were distinguished, based on 4-AP-resistant outward K⁺ currents. IZPC-I had isochronal I_KI current-voltage relations similar to NZPCs whereas IZPC-II showed significantly reduced I_KI and increased outward plateau currents. To study the sustained I_K in the presence of the Class III antiarrhythmic agent E-4031, a two-pulse protocol was used to inactivate transient outward currents, followed by step depolarizations. E-4031-sensitive currents were significantly greater in IZPCs at depolarized potentials (> 0 mV). Similar to NZPCs, IZPC E-4031 currents showed time dependence during depolarization, lack of rectification at positive steps, and voltage-dependent recovery from block.
Conclusion : Decreased I_KI may account for reduced resting potentials in IZPCs. E-4031-sensitive currents in NZPCs, unlike those in canine ventricular myocytes, are sensitive to 4-AP and are larger in IZPCs.     

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     

Influence of Infrasound Exposure on the Whole L-type Calcium Currents in Rat Ventricular Myocytes

This study was designed to examine the effect of infrasound exposure (5 Hz at 130 dB) on whole-cell L-type Ca²⁺ currents (WLCC) in rat ventricular myocytes and the underlying mechanism(s) involved. Thirty-two adult Sprague-Dawley rats were randomly assigned to infrasound exposure and control groups. [Ca²⁺]_i, WLCC, mRNA expression of the a_1c subunit of L-type Ca²⁺ channels (LCC), and SERCA2 protein were examined on day 1, 7, and 14 after initiation of infrasound exposure. Fluo-3/AM fluorescence and the laser scanning confocal microscope techniques were used to measure [Ca²⁺]_i in freshly isolated ventricular myocytes. The Ca²⁺ fluorescence intensity (FI), denoting [Ca²⁺]_i in cardiomyocytes, was significantly elevated in a time-dependent manner in the exposure groups. There was a significant increase in WLCC in the 1-day group and a further significant increase in the 7- and 14-day groups. LCC mRNA expression measured by RT-PCR revealed a significant rise in the 1-day group and a significant additional rise in the 7- and 14-day groups compared with control group. SERCA2 expression was significantly upregulated in the 1-day group followed by an overt decrease in the 7- and 14-day groups. Prolonged exposure of infrasound altered WLCC in rat cardiomyocytes by shifting the steady-state inactivation curves to the right (more depolarized direction) without altering the slope and biophysical properties of I _Ca,L. Taken together, our data suggest that changes in [Ca²⁺]_I levels as well as expression of LCC and SERCA2 may contribute to the infrasound exposure-elicited cardiac response. Zhaohui Pei and Zhiqiang Zhuang contributed equally to this work.     

Temperature Dependence of Macroscopic L-Type Calcium Channel Currents in Single Guinea Pig Ventricular Myocytes

Calcium Channels and Temperature. Introduction: Lowering temperature greatly reduces calcium influx through calcium channels. Studies on a number of tissues demonstrate that the peak inward current, I_Ca exhibits Q₁₀ values ranging from 1.8 to 3.5; however, it remains unclear which component(s) of calcium channel gating may give rise to this large temperature sensitivity. Components of gating that may affect channel availability include phosphorylation and changes in [Ca²⁺)_i, processes that vary in pertinence depending on the channel examined. This study addresses this problem by examining the temperature sensitivity (from 34° to 14°C) of cardiac I_Ca under control conditions, during attenuation or activation of protein kinase A (PKA) activity, and when intracellular [Ca²⁺] has been elevated. Methods and Results: I_Ca was studied using the whole cell configuration of the patch clamp technique. In control, lowering temperature from 34° to 24°C resulted in a shift in the potential for maximum slope (V_a and the peak current (Y_max) toward more positive membrane potentials. The Q₁₀, values for the decrease in Y_max and the macroscopic slope conductance (G_max), which reflects the number of available channels, were 3.15 ± 0.19 and 2.57 ± 0.13, respectively. At 0 mV the Ca²⁺ current decayed biexponentially, and the two time constants (T₁ and T₂) showed Q₁₀ values of 1.79 ± 0.21 and 2.06 ± 038, while their contribution to the total current (I₁ and I₂) showed a Q₁₀ of 5.99 ± 0.83 and 1.61 ± 0.22. In myocytes loaded with inhibitors of the PKA cycle sufficient to inhibit the increase of I_Ca to 1 μM isoprenaline, the Q₁₀ values for some of the kinetic parameters were increased with the Q₁₀ for I₁ increasing to 17.06 ± 3.48. Stimulation of I_Ca by exposing myocytes to 1 μM isoprenaline reduced the temperature sensitivity of Y_max, G_max, and I₁, yielding respective values of 2.00 ± 0.18, 1.85 ± 0.07, and 2.04 ± 0.15. Raising [Ca²⁺], to enhance Ca²⁺_i-dependent inactivation, while affecting inactivation and activation kinetics, affected temperature sensitivity little compared to control. The Q₁₀ for time to peak changed little under experimental conditions (2.3 to 2.4) Conclusions: Increasing the phosphorylated states of calcium channels, but not Ca²⁺_i-dependent inactivation, reduces temperature sensitivity of certain gating parameters. The data suggest that the rate of the transitions between the unavailable and also between the various closed states are changed in the opposite direction to that induced by PKA-dependent phosphorylation. Processes, e.g., inhibitory mechanisms, may be involved to maintain channels in unavailable or “unphosphorylated” states, and it may he these that contribute to the high Q₁₀ of macroscopic channel currents.     

Role of L-Type Calcium Channel Window Current in Generating Current-Induced Early Afterdepolarizations

Ionic Mechanism of EADs. Introduction: Early afterdepolarizations (EADs) can give rise to triggered activity and thereby produce cardiac arrhythmias. We used the whole-cell patch clamp technique to examine the relationship between L-type Ca²⁺ channel window current and the generation of EADs in single ventricular myocytes isolated from guinea pig hearts.
Methods and Results: With a high concentration of EGTA in the internal solution and Na⁺-containing physiologic external solution, EADs were induced in unclamped cells by injecting intracellular depolarizing current pulses. During voltage clamp protocols designed to simulate action potentials interrupted by EADs, we recorded an inward shift in total current up to 0.7 pA/pF over 400 msec at test steps in the range of the take-off potential for EADs. Cd^2t (0.2 mM) blocked most of the inward shift of current during the test steps and abolished EADs. When the same voltage clamp protocol was used following perfusion with an Na⁺-free, K⁺-free external solution, the Cd²⁺-sensitive inward currents recorded during the test steps were similar to those obtained in physiologic external solution. The overlapping range of potentials for partial activation of the d and f variables of L-type Ca²⁺ current ("window" region) measured in Na⁺-free, K⁺-free external solution was virtually the same as the voltage range of the Cd²⁺–sensitive inward currents.
Conclusion: Our experiments suggest that: (1) EADs can arise under conditions of high EGTA buffering of intraccllular [Ca²⁺]; and (2) under these conditions, L-type Ca²⁺ channel window current plays a major role in the initiation of EADs.     

咪达普利对陈旧性心肌梗死代偿区心肌T型钙电流的作用

应用咪达普利(Imi)对家兔陈旧性心肌梗死(OMI)模型进行干预,探讨其对OMI心室肌细胞T型钙电流(ICaT)的影响。选择健康家兔按体重随机分为OMI、假手术组与Imi组。OMI组:采用冠状动脉前降支结扎法制备OMI模型;假手术组:手术与OMI组同,只是不结扎冠状动脉,同样喂养8周;Imi处理组:对OMI家兔口服Imi0.625mg·kg-1·d-1连续8周进行干预。取心脏分离左室游离壁代偿区三层心肌细胞,全细胞膜片钳技术记录ICaT。结果显示,家兔OMI代偿区心肌细胞发生心肌肥厚,细胞膜ICaT密度较假手术组明显增加,当刺激电位为-30mV时,其电流密度从0.35±0.02pA/pF增至2.36±0.12pA/pF(P<0.01)。Imi组代偿区心肌细胞膜ICaT密度降至0.83±0.11pA/pF。ICaT的激活曲线左移,激活曲线斜率增加。Imi组二者改变均减小。但3组心肌细胞ICaT的失活曲线和失活后恢复曲线基本不变。结论:OMI家兔代偿区发生心肌肥厚,ICaT明显增加,Imi可以逆转家兔OMI代偿区心肌肥厚,降低ICaT的电流密度。     

缺氧预处理对缺氧复氧乳鼠心室肌细胞游离钙的影响

观察缺氧预处理对缺氧复氧乳鼠心室肌细胞游离钙的影响。建立培养乳鼠心肌细胞缺氧/复氧损伤模型。设正常对照组(A组)、缺氧复氧组(B组)、缺氧预适应组(C组)。经Flou-3/AM负载染色后,采用流式细胞分析技术,测定细胞内钙离子浓度;利用膜片钳技术,观察L型钙通道和钠钙交换电流的变化。结果:①与A组比较,B组可显著增加细胞内的游离钙离子浓度(P<0.01);L型钙电流密度明显下降,I-V曲线上移,半数失活电压(V1/2)减小,ICa,L失活曲线明显左移,而Na+/Ca2+交换电流显著增加。②与B组比较,C组可减轻缺氧再灌注时[Ca2+]i增加(P<0.01);可减轻再灌注对L型钙电流的抑制,使I-V曲线下移程度减轻,V1/2增加及稳态失活曲线的右移;减少Na+/Ca2+交换电流的增加;③与A组比较,C组钙内流和Na+/Ca2+交换电流有轻度增加(P<0.05)。结论:缺氧预处理使缺氧/复氧造成的[Ca2+]i增高的程度减轻,是通过抑制Na+/Ca2+交换电流的增加实现的。     

缬草单萜氧化物对兔单个心室肌细胞L-型钙电流的影响

利用全细胞膜片钳记录技术研究30μg/L和100μg/L缬草单萜氧化物(VMO)对兔单个心室肌细胞L型钙电流(ICaL)和动作电位的影响。结果:30μg/L和100μg/L的VMO使兔心室肌细胞ICaL峰值由6.04±0.59pA/pF分别减至3.99±0.31pA/pF和2.31±0.24pA/pF(n=8,P<0.01);VMO使ICaL的电流电压曲线上移,但不改变其激活电位、电位峰值和反转电位;VMO还使钙电流失活曲线左移。30μg/LVMO可使动作电位时程(APD)明显缩短,APD50和APD90分别缩短了50.3%和29.6%(n=16,P<0.05),而静息电位和动作电位幅值无明显改变。结论:VMO对LCaL具有浓度依赖性阻滞作用。这可能是其对心血管作用的重要机制之一。     

钾流在糖尿病大鼠心室肌细胞的改变机制

目的研究糖尿病对大鼠心室肌细胞瞬间外向钾流(I_(to))的影响及其分子机制,探讨糖尿病引起的心脏损害与心律失常的关系。方法取体质量150～200 g 的雄性 Sprague-Dawley 大鼠,单次腹腔注射链脲菌素(STZ,65 mg/kg,pH=4.5)建立糖尿病大鼠模型,采用酶解法获得单个心室肌细胞,应用膜片钳全细胞方法记录I_(to);并用反转录聚合酶链式反应技术进一步半定量编码该电流通道α亚单位基因(Kv4.2、Kv4.3和 Kv1.4)mR-NA 的表达水平。结果与对照组比较,+70 mV 时,糖尿病大鼠心室肌细胞 I_(to)密度显著降低[对照组:(30.6±3.8)比糖尿病组:(18.9±3.3)pA/pF,P<0.01);半定量分析法显示糖尿病大鼠心室肌细胞 I_(to)通道α亚单位编码基因 Kv4.2、Kv4.3 mRNA 表达水平分别下调56.9%和46.6%;而 Kv1.4 mRNA 表达则上调约48.0%,3组基因表达水平的改变差异均有统计学意义(P<0.05)。结论糖尿病大鼠心室肌细胞 I_(to)密度显著降低主要与编码该通道α亚单位的基因表达下调有关。     

陈旧性心肌梗死对家兔动作电位时程跨室壁离散及L型钙电流的影响

探讨陈旧性心肌梗死 (MI) (MI后 3个月 ) ,远离MI中心区的心肌细胞电活动的改变。结扎家兔左前降支造成MI模型 ,3个月后酶解分离左室游离壁远离MI中心区的三层 (Epi、M、Endo)心肌细胞 ,采用全细胞膜片钳技术记录单细胞动作电位 (AP)和L型钙电流 (ICa ,L) ,并观察三层心肌细胞AP时程 (APD)的离散性 (TD APD)变化。结果 :MI后 3个月 ,远离MI区的三层心肌细胞的APD明显延长 ,其中Endo心肌细胞的APD明显短于Epi和M心肌细胞(P <0 .0 1)。陈旧性MI组 (OMI)与假手术组 (Sham)及对照组 (Control)比较 ,TD APD显著增加 (2 88.32± 19.5 6vs2 2 8.4 5± 13.94 ,2 10 .32± 17.4 3ms ,P <0 .0 1)。且在OMI组 ,ICa ,L的幅值增加 ,但密度降低 ,其中Endo的ICa,L密度降低最明显 (+10mV时 ,从 16 .12± 1.6 0降至 10 .73± 0 .0 6pA/pF ,降低了 33.4 3% ,Epi从 16 .5 9± 0 .5 0降至 11.75±0 .6 9pA/pF ,降低了 2 9.17% ,M细胞从 18.70± 1.0 3降至 13.2 7± 1.0 5pA/pF ,降低了 2 9.0 3% ,P <0 .0 5 )。结论 :MI3个月后远离MI区的心肌细胞ICa ,L密度明显降低 ,且以Endo降低最明显 ,三层心肌细胞的APD明显延长 ,并且En do心肌细胞的APD明显短于Epi和M心肌细胞 ,TD APD明显增加     

T-Type Calcium Channel Blockade in the Management of Chronic Ischemic Heart Disease

The T-Type calcium channel offers a new therapeutic target for teatment of patients with cardiovascular disease. Mibefradil, a T channel blocker, produces heart rate slowing and coronary vasodilatation but without the negative inotropic effect commonly seen when L-Type channel blockers are used. The present study shows Mibefradil prevents ischemic episodes that are and are not preceded by an increase in heart rate. Although Mibefradil has been withdrawn because of multiple drug interactions, new T-Type calcium channel blockers are under development.