交泰丸含药血清对豚鼠心室肌细胞ICa-L的影响
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摘要
[目的]观察交泰丸及其拆方含药血清对豚鼠心室肌细胞L-型钙离子通道(ICa-L)的影响。[方法]急性分离心室肌细胞,实验分为空白组、黄连组、肉桂组、黄连加肉桂组和交泰丸组,各组加入含药血清浓度为10%和15%,后放于37℃、5%CO_2的孵育箱中孵育24 h,运用膜片钳技术观察药物对ICa-L电流密度和动力学参数的影响。[结果]各组均不同程度抑制钙离子电流(P<0.05),使ICa-L激活和失活时间常数及失活后恢复时间延长(P<0.05)。其中交泰丸对ICa-L平均电流密度抑制率最高达57.5%,肉桂显著延长ICa-L激活时间常数和失活时间常数,黄连使ICa-L失活后恢复时间常数减小最显著。[结论]交泰丸及各拆方含药血清对钙离子通道有不同程度的抑制作用,交泰丸抑制率最大,抑制钙电流影响心肌细胞有效不应期,这可能是交泰丸在临床及实验中具有抗心律失常作用的机制。
[Objective] To explore the effect of Jiaotai Pills and its disassembled prescription in drug serum on ICa-L in guinea-pig ventricular myocytes. [Methods] Acute isolated guinea pig ventricular muscle cells and explore the effect of drug on ICa-L current and and channel dynamics parameters. The myocytes were randomly divided into five groups: the control group, the Rhizoma Coptidis group(decoction of 100 g goldthread), the Cortex Cinnamomi Cassiae group(decoction of 10 g cinnamon), the Rhizoma Coptidis and Cortex Cinnamomi Cassiae group(single decoction of 100 g goldthread + equal volume decoction of 10 g cinnamon), the Jiaotai Pills group(codecoction of 100 g goldthread and 10 g cinnamon). Cells were divided into 10% and 15% drug concentration groups and incubated at 37 ℃ in ahumidified, 5% CO_2 incubator after 24 h, patch clamp test was performed. [Results] Compared with the control group, the drug groups(10%, 15%) inhibited ICa-L density to different degrees(P<0.05), and the highest inhibition rate of 10% concentration Jiaotai Pills was 57.5%. All groups made the activation, inactivation and recovery time contant prolonged(P <0.05). The rate of activation and inactivation time constant of Cortex Cinnamomi Cassiae group was the largest, and the rate of recovery time constant of Rhizoma Coptidis group was the largest. [Conclusion] Drug containing serum of Jiaotai Pills and disassembled prescription have different degrees of inhibition on calcium ion channels, and the mechanism of action are same with calcium blockers, it could be the mechanism of Jiaotai Pills antiarrhythmic.
[Objective] To explore the effect of Jiaotai Pills and its disassembled prescription in drug serum on ICa-L in guinea-pig ventricular myocytes. [Methods] Acute isolated guinea pig ventricular muscle cells and explore the effect of drug on ICa-L current and and channel dynamics parameters. The myocytes were randomly divided into five groups: the control group, the Rhizoma Coptidis group(decoction of 100 g goldthread), the Cortex Cinnamomi Cassiae group(decoction of 10 g cinnamon), the Rhizoma Coptidis and Cortex Cinnamomi Cassiae group(single decoction of 100 g goldthread + equal volume decoction of 10 g cinnamon), the Jiaotai Pills group(codecoction of 100 g goldthread and 10 g cinnamon). Cells were divided into 10% and 15% drug concentration groups and incubated at 37 ℃ in ahumidified, 5% CO_2 incubator after 24 h, patch clamp test was performed. [Results] Compared with the control group, the drug groups(10%, 15%) inhibited ICa-L density to different degrees(P<0.05), and the highest inhibition rate of 10% concentration Jiaotai Pills was 57.5%. All groups made the activation, inactivation and recovery time contant prolonged(P <0.05). The rate of activation and inactivation time constant of Cortex Cinnamomi Cassiae group was the largest, and the rate of recovery time constant of Rhizoma Coptidis group was the largest. [Conclusion] Drug containing serum of Jiaotai Pills and disassembled prescription have different degrees of inhibition on calcium ion channels, and the mechanism of action are same with calcium blockers, it could be the mechanism of Jiaotai Pills antiarrhythmic.
引文
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[9]丽英,杨艳敏,浦介麟,等.盐酸关附甲素对豚鼠和大鼠心肌细胞钾通道的阻断作用[J].中国心脏起搏与心电生理杂志,2006,20(3):255-258.
[10]Collins TP,Terrar DA.Ca2+-stimulated adenylyl cyclases regulate the L-type Ca2+current in guinea-pig atrial myocytes[J].J Physiol,2012,590(8):1881-1893.
[11]姚岚,方芳.作用于心肌细胞离子通道的抗心律失常中药研究进展[J].中西医结合心脑血管病杂志,2017,15(9):1057-1059.
[12]孙丽.浅谈中药制剂抗心律失常作用研究进展[J].中国现代药物应用,2017,11(10):194-195.
[13]谢凌鹏,袁立霞,周楚莹,等.桂枝甘草配伍治疗心律失常浅析[J].江苏中药,2015,47(4):57-58.
[14]朱明军,刘红军,王永霞,等.心悸宁浸膏粉溶液对豚鼠心室肌细胞动作电位的影响[J].中国实验方剂学杂志,2008,14(8):45-47.
[15]Tolkacheva EG,Anumonwo JM,Jalife J.Action potential duration restitution portraits of mammalian ventricular myocytes:role of calcium current[J].Biophys J,2006,91(7):2735-2745.
[16]Limpitikul WB,Dick IE,Joshi-Mukherjee R,et al.Calmodulin mutations associated with long QT syndrome prevent inactivation of cardiac L-type Ca2+currents and promote proarrhythmic behavior in ventricular myocytes[J].J Mol Cell Cardiol,2014,74:115-124.
[17]Viola HM,Jordan MC,Roos KP,et al.Decreased myocardial injury and improved contractility after administration of a peptide derived against the alpha-interacting domain of the L-type calcium channel[J].J Am Heart Assoc,2014,3(3):e000961.
[2]Yasuda J,Okada M,Yamawaki H.Endostatin inhibits T-type Ca2+channel current in guinea pig ventricular myocyte[J].J Vet Med Sci.2015;77(10):1289-1291.
[3]Ji J,Kang J,Rampe D.L-type Ca2+channel responses to bay k 8644in stem cell-derived cardiomyocytes are unusually dependent on holding potential and charge carrier[J].Assay Drug Dev Technol,2014,12(6):352-360.
[4]Frolov RV,Singh S.Evidence of more ion channels inhibited by celecoxib:KV1.3 and L-type Ca2+channels[J].BMC Res Notes,2015:8,62.
[5]曹英杰,原双兴,高原,等.交泰丸指纹图谱研究[J].中国医院药学杂志,2015,35(10):920-922.
[6]Thomas P,Collins and Derek A.Terrar.Ca2+-stimulated adenylyl cyclases regulate the L-type Ca2+current in guinea-pig atrial myocytes[J].The Journal of Physiology,2012;590(8):1881-1893.
[7]Marvin WJ,Jr,Chittick VL,Rosenthal JK,et al.The Isolated Sinoatrial Node Cell in Primary Culture from the Newborn Rat[J].Circulation Research,1984;55(2):252-260.
[8]周彤,毛晓波,毛奕,等.细胞外液成分对耐钙心肌细胞急性分离及电生理记录的影响[J].山西医科大学学报,2009,40(5):393-397.
[9]丽英,杨艳敏,浦介麟,等.盐酸关附甲素对豚鼠和大鼠心肌细胞钾通道的阻断作用[J].中国心脏起搏与心电生理杂志,2006,20(3):255-258.
[10]Collins TP,Terrar DA.Ca2+-stimulated adenylyl cyclases regulate the L-type Ca2+current in guinea-pig atrial myocytes[J].J Physiol,2012,590(8):1881-1893.
[11]姚岚,方芳.作用于心肌细胞离子通道的抗心律失常中药研究进展[J].中西医结合心脑血管病杂志,2017,15(9):1057-1059.
[12]孙丽.浅谈中药制剂抗心律失常作用研究进展[J].中国现代药物应用,2017,11(10):194-195.
[13]谢凌鹏,袁立霞,周楚莹,等.桂枝甘草配伍治疗心律失常浅析[J].江苏中药,2015,47(4):57-58.
[14]朱明军,刘红军,王永霞,等.心悸宁浸膏粉溶液对豚鼠心室肌细胞动作电位的影响[J].中国实验方剂学杂志,2008,14(8):45-47.
[15]Tolkacheva EG,Anumonwo JM,Jalife J.Action potential duration restitution portraits of mammalian ventricular myocytes:role of calcium current[J].Biophys J,2006,91(7):2735-2745.
[16]Limpitikul WB,Dick IE,Joshi-Mukherjee R,et al.Calmodulin mutations associated with long QT syndrome prevent inactivation of cardiac L-type Ca2+currents and promote proarrhythmic behavior in ventricular myocytes[J].J Mol Cell Cardiol,2014,74:115-124.
[17]Viola HM,Jordan MC,Roos KP,et al.Decreased myocardial injury and improved contractility after administration of a peptide derived against the alpha-interacting domain of the L-type calcium channel[J].J Am Heart Assoc,2014,3(3):e000961.