Objective：To study the effects of sodium ferulate on the ultrarapid delayed rectifier K^＋ current（IKur） in human atrial myocytes. Methods：Human atrial myocytes were isolated by enzyme dispersion method. IKur, in ...Objective：To study the effects of sodium ferulate on the ultrarapid delayed rectifier K^＋ current（IKur） in human atrial myocytes. Methods：Human atrial myocytes were isolated by enzyme dispersion method. IKur, in human atrial myocytes were recorded by using the whole cell patch clamp. The changes of IKur were compared in the absence and the presence of sodium ferulate. Results：There was no effect of 0.4 g/L sodium ferulate on I-V relation of IKur. However, 0.4 g/L sodium ferulate inhibited IKur to some degrees at each test pulse. The current densities of IKur at ＋60 mV decreased from 4.997 ± 0.35 PA/PF to 3.331 ± 0.26 PA/PF（n = 6, P 〈 0.05）. The inhibitory effect was concentration-dependent. IC50 was（0.41 ±0.03）g/L and the Hill coefficient was 0.95 ± 0.05. Conclusion：Sodium ferulate as a potassium channel blocker can inhibit IKur in human atrial myocytes effectively.展开更多
Aim Evidence has shown that stimulation of alA-adrenorecetors receptor （alA-AR） or angiotensin II type 1 receptor （AT1R） acutely down-regulates the rapid component of the delayed rectifier K ＋ current （IKr） via...Aim Evidence has shown that stimulation of alA-adrenorecetors receptor （alA-AR） or angiotensin II type 1 receptor （AT1R） acutely down-regulates the rapid component of the delayed rectifier K ＋ current （IKr） via protein kinase C （PKC）. This study was designed to investigate which PKC isozymes mediate down-regulations of IKr by alA-AR and AT1R. Method The whole-cell patch-clamp technique was used to record IKr in native cardio- myocytes and in human embryonic kidney （HEK） 293 cells co-transfected with human ether-a-go-go related gene （hERG） encoding α-subunit of IKr and human alA-AR or AT1R gene. Result In isolated guinea-pig ventricular cardiomyocytes the inhibitory action of Ang II on IKr was little affected by Go6976 （selectively inhibiting PKCα, β and γ） and Go6983 （selectively inhibiting PKCα, β, γ , δ, and ζ）, but was significantly antagonized by an inter- nal dialysis with PKCe-selective inhibitory peptide εV1 -2. In contrast, the inhibitory action of alA-AR agonist A61603 on IKr was remarkably attenuated by Go6976 or Go6983, but not affected by peptide εV1 -2. Moreover, specific PKC-selective inhibitory peptide antagonized the effect of A61603. The results suggested that PKCe and PKCα isoform respectively mediated the inhibitory effect of AT1R and a1A-AR. In heterologous expression system, both PKCα and e-selective activator peptides down regulated hERG current with different manner. PKCα activator peptide shifted the activation curve of the channel to the right, but PKCe-selective activator peptide did not. Simi- larly, A61603 shifted the activation curve to the right, whereas Ang Ⅱ had no effect. In addition, both A61603 and PKCα activator peptide showed inhibitory action on bERG A PKC current （an bERG mutant in which 17 of the 18 ROSITE-predicted PKC acceptor serines/threonines were changed to alanine） with a similar potency to wild type bERG current. But, both Ang Ⅱ and PKCe-selective activator peptide exhibited no effects on bERG △ PKC cur- rent. The results indicated that PKCα and PKCe isoforms down-regulated bERG current through different mecha- nism. Conclusion PKCα and PKCe isoform respectively mediates the inhibition on IKr by stimulation of AT1R and alA-AR via different molecular mechanism.展开更多
The aim of this study was to compare the effects of d, l-Sotalol and dSotalol on the delayed rectifier K+ outward current in the presence of isoproterenol at different concentrations. Time-dependent delayed rectifier...The aim of this study was to compare the effects of d, l-Sotalol and dSotalol on the delayed rectifier K+ outward current in the presence of isoproterenol at different concentrations. Time-dependent delayed rectifier K+ outward currents were measured in isolated guinea pig single myocytes using the whole-cell configuration of the patch-clamp technique. Currents were measured in response to 300 ms depolarizing pulses from a holding potential of -40 mV in three experimental protocols [control, isoproterenol (10^(9)mol/L - 10^(-6) mol/L ), and isoproterenol (10^(-9)mol/L - 10^(-6)mol/L ) plus either d, l-Sotalol (10^(-4) mol/L) or d-Sotalol (10^(-4) mol/L)]. IK tail currents were measured upon repolarization to -40 mV. It was found that Ik was significantly amplified in the presence. of isoproterenol (10^(-9) mol/L- 10^(-6) mol/L) plus d-Sotalol. At 10-8 mol/L isoproterenol, Ik was increased by 92. 7%±17. 1 % (P<0. 05) and 54. 3 %±13. 4 % after d-Sotalol addition (P<0. 05). In contrast, d, l-Sotalol completely conteracted the increase of iK by isoproterenol (<10^(-8) mol/L), and compared to control, Ic was decreased by 35. 6 % ±8. 1% at 10^(-8) mol/L isoproterenol plus d, l-Sotalol (P<0. 05). It is concluded that the β-adrenergic blocking property of d, l-Sotalol but not that of dSotalol maintains the delayed rectifier K+ outward current blockade in the presence of isoproterenol in guinea pig myocytes. This might contribute to a superior antiarrhythmic efficacy as compared to d-Sotalol.展开更多
To investigate the effect of intedeukin-1β （IL-1β） on IA and IK currents in cultured murine trigeminal ganglion （TG） neurons, whole-cell patch clamp technique was used to record the IA and IK currents before and...To investigate the effect of intedeukin-1β （IL-1β） on IA and IK currents in cultured murine trigeminal ganglion （TG） neurons, whole-cell patch clamp technique was used to record the IA and IK currents before and after 20 ng/mL IL-1β perfusion. Our results showed that 20 ng/mL IL-1β inhibited IA currents （18.3±10.7）% （n=6, P〈0.05）. IL-1β at 20 ng/mL had no effect on G-V curve of IA but moved the H-infinity curve V0.5 from -36.6±6. 1 mV to-42.4±5.2 mV （n=5, P〈0.01）. However, 20 ng/mL IL-1β had effect on neither the amplitude nor the G-V curve of IK. IL-1β was found to selectively inhibit IA current in TG neurons and the effect may contribute to hyperalgesia under various inflammatory conditions.展开更多
Objective To investigate changes in the delayed rectifier K + channel (Kv) function and the regulation of Kv by the protein kinase C (PKC) pathway in bronchial myocytes from asthmatic rats. Methods The Kv currents an...Objective To investigate changes in the delayed rectifier K + channel (Kv) function and the regulation of Kv by the protein kinase C (PKC) pathway in bronchial myocytes from asthmatic rats. Methods The Kv currents and membrane potentials in bronchial myocytes from asthmatic rats and from controls were observed, using whole cell voltage- and current-patch clamp techniques.Results Bronchial myocytes from asthmatic rats showed a significant reduction in Kv-current density (51.6±9.4 pA/pF, n=14, P<0.01) in comparison with those from control rats (72.4±12.3 pA/pF, n=14) at +50 mV. The current-voltage relationship curve exhibited a significant downward shift. Bronchial myocytes from asthmatic rats had no significantly different capacitances (P>0.05), but had more positive membrane potential ( P<0.01) compared with those from controls. 1 μmol/L phorbol 12-myristate 13-acetate, a PKC activator, caused an obvious reduction in Kv-current density (P<0.01) and a significant downward shift in the current-voltage relationship curve, an effect which was partly abolished by 1 μmol/L Ro31-8220 (a PKC inhibitor); 1 μmol/L phorbol 12-myristate 13-acetate caused more positive membrane potential (Em), from -36.8±5.7 mV to -30.4±7.3 mV, in rat bronchial myocytes (P<0.05). This effect was partly abolished by 1 μmol/L Ro31-8220. Conclusions Bronchial myocytes from asthmatic rats have inhibited Kv function, more positive membrane potential, and higher excitability, all of which can also be induced by PKC activation. These characteristics may contribute to the development of airway hyperreactivity in asthma.展开更多
Objective Catecholamines antagonize the clinical efficacy of pure class Ⅲ antiarrhythmic agents in vivo. The antiarrhythmic agent d, l sotalol has β adrenergic blocking properties and class Ⅲ activity. However, ...Objective Catecholamines antagonize the clinical efficacy of pure class Ⅲ antiarrhythmic agents in vivo. The antiarrhythmic agent d, l sotalol has β adrenergic blocking properties and class Ⅲ activity. However, its d isomer without β blockade has been shown to exert significant proarrhythmia. To determine the role of β adrenergic blocking properties of d, l sotalol on its antiarrhythmic effect, we compared the effects of d, l sotalol and d sotalol on delayed rectifier K + outward current in the presence of isoproterenol at different concentrations. Methods Time dependent delayed rectifier K + outward currents, I K (I Kr and I Ks ) and tail current (I K tail ) were measured in isolated guinea pig myocytes using the whole cell configuration of the patch clamp technique. Currents were measured in response to 300 ms depolarizing pulses from a holding potential of Department of Cardiology, University Hospital Heidelberg, Germany (Yao XZ, Yannoulis NC, Kiehn J and Brachmann J) 40 mV in three experimental protocols [control, isoproterenol (10 9 -10 6 mol/L), and isoproterenol (10 9 -10 6 mol/L) plus either d, l sotalol (10 4 mol/L) or d sotalol (10 4 mol/L)]. I K tail currents were measured upon repolarization to 40 mV. Results Isoproterenol significantly inreased I K and I K tail in a concentration dependent manner. I K was significantly amplified in the presence of isoproterenol (10 9 -10 6 mol/L) plus d sotalol. At 10 8 mol/L isoproterenol, I K was increased by 92.3%±23.7% before and 54.3%±13.4% after d sotalol. In contrast, d, l sotalol strongly suppressed the effect of isoproterenol on I K, and compared to control, I K was decreased by 35.6%±8.1% at 10 8 mol/L isoproterenol. Conclusions The β adrenergic blocking property of d, l sotalol maintains delayed rectifier K + outward current block in the presence of isoproterenol in guinea pig myocytes. This may result in its supperior antiarrhythmic efficacy compared to d sotalol.展开更多
基金This work was supported by the National Natural Science Foundation of China(No.30700747)
文摘Objective：To study the effects of sodium ferulate on the ultrarapid delayed rectifier K^＋ current（IKur） in human atrial myocytes. Methods：Human atrial myocytes were isolated by enzyme dispersion method. IKur, in human atrial myocytes were recorded by using the whole cell patch clamp. The changes of IKur were compared in the absence and the presence of sodium ferulate. Results：There was no effect of 0.4 g/L sodium ferulate on I-V relation of IKur. However, 0.4 g/L sodium ferulate inhibited IKur to some degrees at each test pulse. The current densities of IKur at ＋60 mV decreased from 4.997 ± 0.35 PA/PF to 3.331 ± 0.26 PA/PF（n = 6, P 〈 0.05）. The inhibitory effect was concentration-dependent. IC50 was（0.41 ±0.03）g/L and the Hill coefficient was 0.95 ± 0.05. Conclusion：Sodium ferulate as a potassium channel blocker can inhibit IKur in human atrial myocytes effectively.
文摘Aim Evidence has shown that stimulation of alA-adrenorecetors receptor （alA-AR） or angiotensin II type 1 receptor （AT1R） acutely down-regulates the rapid component of the delayed rectifier K ＋ current （IKr） via protein kinase C （PKC）. This study was designed to investigate which PKC isozymes mediate down-regulations of IKr by alA-AR and AT1R. Method The whole-cell patch-clamp technique was used to record IKr in native cardio- myocytes and in human embryonic kidney （HEK） 293 cells co-transfected with human ether-a-go-go related gene （hERG） encoding α-subunit of IKr and human alA-AR or AT1R gene. Result In isolated guinea-pig ventricular cardiomyocytes the inhibitory action of Ang II on IKr was little affected by Go6976 （selectively inhibiting PKCα, β and γ） and Go6983 （selectively inhibiting PKCα, β, γ , δ, and ζ）, but was significantly antagonized by an inter- nal dialysis with PKCe-selective inhibitory peptide εV1 -2. In contrast, the inhibitory action of alA-AR agonist A61603 on IKr was remarkably attenuated by Go6976 or Go6983, but not affected by peptide εV1 -2. Moreover, specific PKC-selective inhibitory peptide antagonized the effect of A61603. The results suggested that PKCe and PKCα isoform respectively mediated the inhibitory effect of AT1R and a1A-AR. In heterologous expression system, both PKCα and e-selective activator peptides down regulated hERG current with different manner. PKCα activator peptide shifted the activation curve of the channel to the right, but PKCe-selective activator peptide did not. Simi- larly, A61603 shifted the activation curve to the right, whereas Ang Ⅱ had no effect. In addition, both A61603 and PKCα activator peptide showed inhibitory action on bERG A PKC current （an bERG mutant in which 17 of the 18 ROSITE-predicted PKC acceptor serines/threonines were changed to alanine） with a similar potency to wild type bERG current. But, both Ang Ⅱ and PKCe-selective activator peptide exhibited no effects on bERG △ PKC cur- rent. The results indicated that PKCα and PKCe isoforms down-regulated bERG current through different mecha- nism. Conclusion PKCα and PKCe isoform respectively mediates the inhibition on IKr by stimulation of AT1R and alA-AR via different molecular mechanism.
文摘The aim of this study was to compare the effects of d, l-Sotalol and dSotalol on the delayed rectifier K+ outward current in the presence of isoproterenol at different concentrations. Time-dependent delayed rectifier K+ outward currents were measured in isolated guinea pig single myocytes using the whole-cell configuration of the patch-clamp technique. Currents were measured in response to 300 ms depolarizing pulses from a holding potential of -40 mV in three experimental protocols [control, isoproterenol (10^(9)mol/L - 10^(-6) mol/L ), and isoproterenol (10^(-9)mol/L - 10^(-6)mol/L ) plus either d, l-Sotalol (10^(-4) mol/L) or d-Sotalol (10^(-4) mol/L)]. IK tail currents were measured upon repolarization to -40 mV. It was found that Ik was significantly amplified in the presence. of isoproterenol (10^(-9) mol/L- 10^(-6) mol/L) plus d-Sotalol. At 10-8 mol/L isoproterenol, Ik was increased by 92. 7%±17. 1 % (P<0. 05) and 54. 3 %±13. 4 % after d-Sotalol addition (P<0. 05). In contrast, d, l-Sotalol completely conteracted the increase of iK by isoproterenol (<10^(-8) mol/L), and compared to control, Ic was decreased by 35. 6 % ±8. 1% at 10^(-8) mol/L isoproterenol plus d, l-Sotalol (P<0. 05). It is concluded that the β-adrenergic blocking property of d, l-Sotalol but not that of dSotalol maintains the delayed rectifier K+ outward current blockade in the presence of isoproterenol in guinea pig myocytes. This might contribute to a superior antiarrhythmic efficacy as compared to d-Sotalol.
基金The project was supported by a grant from the National Natural Sciences Foundation of China (No. 30271500)
文摘To investigate the effect of intedeukin-1β （IL-1β） on IA and IK currents in cultured murine trigeminal ganglion （TG） neurons, whole-cell patch clamp technique was used to record the IA and IK currents before and after 20 ng/mL IL-1β perfusion. Our results showed that 20 ng/mL IL-1β inhibited IA currents （18.3±10.7）% （n=6, P〈0.05）. IL-1β at 20 ng/mL had no effect on G-V curve of IA but moved the H-infinity curve V0.5 from -36.6±6. 1 mV to-42.4±5.2 mV （n=5, P〈0.01）. However, 20 ng/mL IL-1β had effect on neither the amplitude nor the G-V curve of IK. IL-1β was found to selectively inhibit IA current in TG neurons and the effect may contribute to hyperalgesia under various inflammatory conditions.
基金ThisstudywassupportedbytheNationalNaturalScienceFoundationofChina (No 3 0 2 70 5 83 ) .
文摘Objective To investigate changes in the delayed rectifier K + channel (Kv) function and the regulation of Kv by the protein kinase C (PKC) pathway in bronchial myocytes from asthmatic rats. Methods The Kv currents and membrane potentials in bronchial myocytes from asthmatic rats and from controls were observed, using whole cell voltage- and current-patch clamp techniques.Results Bronchial myocytes from asthmatic rats showed a significant reduction in Kv-current density (51.6±9.4 pA/pF, n=14, P<0.01) in comparison with those from control rats (72.4±12.3 pA/pF, n=14) at +50 mV. The current-voltage relationship curve exhibited a significant downward shift. Bronchial myocytes from asthmatic rats had no significantly different capacitances (P>0.05), but had more positive membrane potential ( P<0.01) compared with those from controls. 1 μmol/L phorbol 12-myristate 13-acetate, a PKC activator, caused an obvious reduction in Kv-current density (P<0.01) and a significant downward shift in the current-voltage relationship curve, an effect which was partly abolished by 1 μmol/L Ro31-8220 (a PKC inhibitor); 1 μmol/L phorbol 12-myristate 13-acetate caused more positive membrane potential (Em), from -36.8±5.7 mV to -30.4±7.3 mV, in rat bronchial myocytes (P<0.05). This effect was partly abolished by 1 μmol/L Ro31-8220. Conclusions Bronchial myocytes from asthmatic rats have inhibited Kv function, more positive membrane potential, and higher excitability, all of which can also be induced by PKC activation. These characteristics may contribute to the development of airway hyperreactivity in asthma.
文摘Objective Catecholamines antagonize the clinical efficacy of pure class Ⅲ antiarrhythmic agents in vivo. The antiarrhythmic agent d, l sotalol has β adrenergic blocking properties and class Ⅲ activity. However, its d isomer without β blockade has been shown to exert significant proarrhythmia. To determine the role of β adrenergic blocking properties of d, l sotalol on its antiarrhythmic effect, we compared the effects of d, l sotalol and d sotalol on delayed rectifier K + outward current in the presence of isoproterenol at different concentrations. Methods Time dependent delayed rectifier K + outward currents, I K (I Kr and I Ks ) and tail current (I K tail ) were measured in isolated guinea pig myocytes using the whole cell configuration of the patch clamp technique. Currents were measured in response to 300 ms depolarizing pulses from a holding potential of Department of Cardiology, University Hospital Heidelberg, Germany (Yao XZ, Yannoulis NC, Kiehn J and Brachmann J) 40 mV in three experimental protocols [control, isoproterenol (10 9 -10 6 mol/L), and isoproterenol (10 9 -10 6 mol/L) plus either d, l sotalol (10 4 mol/L) or d sotalol (10 4 mol/L)]. I K tail currents were measured upon repolarization to 40 mV. Results Isoproterenol significantly inreased I K and I K tail in a concentration dependent manner. I K was significantly amplified in the presence of isoproterenol (10 9 -10 6 mol/L) plus d sotalol. At 10 8 mol/L isoproterenol, I K was increased by 92.3%±23.7% before and 54.3%±13.4% after d sotalol. In contrast, d, l sotalol strongly suppressed the effect of isoproterenol on I K, and compared to control, I K was decreased by 35.6%±8.1% at 10 8 mol/L isoproterenol. Conclusions The β adrenergic blocking property of d, l sotalol maintains delayed rectifier K + outward current block in the presence of isoproterenol in guinea pig myocytes. This may result in its supperior antiarrhythmic efficacy compared to d sotalol.
文摘目的观察枸杞多糖(lycium bararum polysaccharides,LBP)对体外高压诱导调亡的视网膜神经节细胞(retinal ganglion cells,RGCs)钾电流的影响。方法生后2~3 d的SD乳大鼠RGCs原代培养,分为对照组、加压组和LBP组,对照组为常规培养6 d;加压组为常规培养6 d后用自行设计的加压装置加压1 h 80 mm Hg(1 k Pa=7.5 mm Hg);LBP组为常规培养5 d后,加入LBP共培养24 h后,再加压80 mm Hg 1 h。通过全细胞膜片钳技术观察各组钾电流、半数最大激活电压(V1/2)、斜率(K)、最大电导(Gmax)的变化。结果加压能使电流幅度显著增加,LBP能抑制加压引起的电流增加。在刺激电位为-10~60 m V时,加压组的电流密度明显大于对照组,差异有统计学意义(均为P<0.01,n=5/6);LBP组电流密度明显小于加压组,差异有统计学意义(均为P<0.01,n=6/8)。三组钾电流V1/2总体差异有统计学意义(F=55.60,P<0.01),加压组V1/2(11.65±1.30)m V与对照组(21.42±1.33)m V、LBP组(19.33±13.75)m V比较,差异均有统计学意义(均为P<0.01);LBP组V1/2与对照组V1/2比较,差异无统计学意义(P>0.05)。三组K值分别是17.09±1.24、16.58±1.18、18.13±1.29,总体差异无统计学意义(F=1.39,P>0.05)。三组Gmax总体差异有统计学意义(F=3.77,P<0.05),加压组Gmax(0.59±0.13)与对照组Gmax(0.46±0.06)、LBP组Gmax(0.46±0.07)比较,差异均有统计学意义(均为P<0.05);LBP组Gmax与对照组Gmax比较,差异无统计学意义(P>0.05)。结论 LBP能抑制加压引起的RGCs钾电流增加,对RGCs具有保护作用。
