调脉饮及其拆方对快速心律失常大鼠心肌L型钙通道mRNA表达的影响
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摘要
目的:观察调脉饮全方及其拆方对快速性心律失常模型大鼠心电图及心室肌L型钙通道(L-type calcium channels,LTCC)α1c亚基mRNA表达的影响。方法:把60只Wistar大鼠随机分成6组,每组各10只,分别为模型组,调脉饮全方组(22 g·kg~(-1)),凉血清热组(7 g·kg~(-1)),行气通脉组(7.5 g·kg~(-1)),益气养心组(7.5 g·kg~(-1))和西药组(胺碘酮,100 mg·kg~(-1))。除模型组外,每组按相应剂量分别灌胃给药7 d。采用乌头碱舌下静脉注射造模,分别记录室性早搏、室性心动过速和室颤及死亡出现的时间,通过实时荧光定量聚合酶链式反应(Real-time PCR)检测和蛋白免疫印迹法(Western blot)测定各组心室肌组织LTCCα1c亚基mRNA和蛋白表达情况。结果:与模型组比较,全方组、凉血清热组、胺碘酮组大鼠出现室性早搏时间均有延迟,其中全方组、胺碘酮组大鼠出现室性早搏时间延迟明显(P<0.05,P<0.01);全方组、凉血清热组、胺碘酮组出现室性心动过速时间明显延迟(P<0.05);全方组、凉血清热组、胺碘酮组出现室颤时间明显延迟(P<0.05,P<0.01);全方组、凉血清热组、胺碘酮组出现死亡时间明显延迟(P<0.05,P<0.01)。与模型组相比,全方组、凉血清热组、胺碘酮组大鼠心肌LTCCα1c亚基mRNA表达均有所降低,其中全方组和胺碘酮组与模型组相比有显著差异(P<0.05);与模型组比较,全方组、凉血清热组、胺碘酮组大鼠心肌LTCCα1c亚基蛋白表达均有所降低,有显著差异(P<0.01)。结论:调脉饮全方及其中凉血清热药具有抗实验性快速心律失常的作用,作用机制可能与干预心肌组织LTCC mRNA的表达水平有关。
Objective: To observe the effect of the whole prescription and decomposed recipes of Tiaomai drink on the electrocardiogram changes of experimental tachyarrthythmia rats and the mRNA expression levels ofα1 c sub-units in L-type calcium channels( LTCC) of myocardium. Method: The 60 Wistar rats were randomly divided into the model group,Tiaomai drink whole prescription group( 22 g·kg~(-1)),blood cooling and heat clearing medicine group( 7 g·kg~(-1)),Qi-regulating and blood flow promoting medicine group( 7. 5 g·kg~(-1)),Qi and heart tonifying medicine group( 7. 5 g·kg~(-1)),and western medicine group( amiodarone,100 mg·kg~(-1)).Except those in model group,corresponding dose of medicines was given in other groups for 7 days. Sublingual intravenous injection of aconitine was used for modeling,and then the time to have ventricular premature beat,ventricular tachycardia, ventricular fibrillation and death was recorded respectively. Quantitative Real-time polymerase chain reaction( Real-time PCR) and Western blot were respectively used to detect the mRNA and protein expression levels of LTCCα1 c sub-units in rat myocardial tissue. Result: As compared with the model group,there was a delay in ventricular premature beats in the whole prescription group,blood cooling group and the western medicine group,among which the time delay of ventricular premature beat was obvious in the whole prescription group and the western medicine group( P < 0. 005,P < 0. 01); the time of ventricular tachycardia was significantly delayed in the whole prescription group,blood cooling group and western medicine group( P < 0. 05);the time of ventricular fibrillation was significantly delayed in the whole prescription group,blood cooling group and western medicine group( P < 0. 05,P < 0. 01); the death time was significantly delayed in the whole prescription group,blood cooling group and western medicine group( P < 0. 05,P < 0. 01). As compared with the model group,the mRNA expression of myocardial LTCCα1 c was decreased in the whole prescription group,blood cooling group and the western medicine group,and the whole prescription group and the western medicine group were significantly different from the model group( P < 0. 01); the protein expression of myocardial LTCCα1 c was significantly decreased in the whole prescription group,blood cooling group and the western medicine group.Conclusion: The whole prescription as well as blood cooling and heat clearing medicine were effective for experimental tachyarrhythmia,and the mechanism may be related to the intervention on the mRNA expression level of the myocardial tissue L-type calcium channel.
Objective: To observe the effect of the whole prescription and decomposed recipes of Tiaomai drink on the electrocardiogram changes of experimental tachyarrthythmia rats and the mRNA expression levels ofα1 c sub-units in L-type calcium channels( LTCC) of myocardium. Method: The 60 Wistar rats were randomly divided into the model group,Tiaomai drink whole prescription group( 22 g·kg~(-1)),blood cooling and heat clearing medicine group( 7 g·kg~(-1)),Qi-regulating and blood flow promoting medicine group( 7. 5 g·kg~(-1)),Qi and heart tonifying medicine group( 7. 5 g·kg~(-1)),and western medicine group( amiodarone,100 mg·kg~(-1)).Except those in model group,corresponding dose of medicines was given in other groups for 7 days. Sublingual intravenous injection of aconitine was used for modeling,and then the time to have ventricular premature beat,ventricular tachycardia, ventricular fibrillation and death was recorded respectively. Quantitative Real-time polymerase chain reaction( Real-time PCR) and Western blot were respectively used to detect the mRNA and protein expression levels of LTCCα1 c sub-units in rat myocardial tissue. Result: As compared with the model group,there was a delay in ventricular premature beats in the whole prescription group,blood cooling group and the western medicine group,among which the time delay of ventricular premature beat was obvious in the whole prescription group and the western medicine group( P < 0. 005,P < 0. 01); the time of ventricular tachycardia was significantly delayed in the whole prescription group,blood cooling group and western medicine group( P < 0. 05);the time of ventricular fibrillation was significantly delayed in the whole prescription group,blood cooling group and western medicine group( P < 0. 05,P < 0. 01); the death time was significantly delayed in the whole prescription group,blood cooling group and western medicine group( P < 0. 05,P < 0. 01). As compared with the model group,the mRNA expression of myocardial LTCCα1 c was decreased in the whole prescription group,blood cooling group and the western medicine group,and the whole prescription group and the western medicine group were significantly different from the model group( P < 0. 01); the protein expression of myocardial LTCCα1 c was significantly decreased in the whole prescription group,blood cooling group and the western medicine group.Conclusion: The whole prescription as well as blood cooling and heat clearing medicine were effective for experimental tachyarrhythmia,and the mechanism may be related to the intervention on the mRNA expression level of the myocardial tissue L-type calcium channel.
引文
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[15]张林,王停,杜守颖,等.芍药苷、薄荷脑对葛根素在Calu-3细胞模型转运过程中膜流动性、钠-钾离子ATP酶和钙离子ATP酶的影响[J].中国中药杂志,2018,43(4):731-735.
[16]刘和兰,莫丹,梁荣寿,等.芍药苷对异丙肾上腺素诱导大鼠心肌肥大的抑制作用及其机制研究[J].中国实验方剂学杂志,2015,21(16):88-92.
[17]张广钦,郝雪梅,陈世忠,等.芍药苷对大鼠心肌细胞L钙通道的阻断作用[J].中国药理学通报,2003,19(8):863-866.
[18]刘丹,曹广尚,司席席,等.黄连中生物碱类成分抗心律失常研究概述[J].山东中医杂志,2017,36(2):164-171.
[19]徐尚忠,张翼,任建英,等.小檗碱对豚鼠心室肌细胞L-及T-型钙离子通道的影响[J].中国药理学报,1997,18(6):515-518.
[20]司晓晨,R.Clintton Webb.黄连素片对钙离子通道的拮抗作用[J].南京中医药大学学报,1996,12(6):20-22.
[21]魏婷,梁喆,金彦,等.小檗碱、莲心碱和甲基莲心碱对HERG通道表达的影响[J].中国中药杂志,2013,38(2):239-244.
[22]Furukawa Y,XUE Y X,Chiba S,et al.Effects of zatebradine on ouabain,two-stage coronary ligation and epinephrine-induced ventricular tachyarrhythmias[J].Eur J Pharmacol,1996,300(3):203-210.
[23]Faber E M,Rudy Y.Action potential and contraetility changes in[Na+]ioverloaded cardiac myocytes:a simulation study[J].Bio Phys J,2000,78:2392-2404.
[24]Friese J,Gleitz J,Gutser U T,et al.Aconitum sp.alkaloids:the modulation of voltage-dependent Na+channels,toxieity and antinociceptive properties[J].Eur J Pharmaeol,1997,337(2/3):165-174.
[25]刘影,单宏丽,孙宏丽,等.乌头碱对大鼠心室肌细胞包浆钙及L-型钙电流的影响[C]∥中国病理生理学会动脉粥样硬化专业委员会.中国病理生理学会动脉粥样硬化专业委员会五届一次会议论文集:2002卷.衡阳:中国动脉硬化杂志,2002:101-102.
[26]龚冬梅,单宏丽,周宇宏,等.哇巴因和乌头碱诱发豚鼠和大鼠心律失常的离子作用靶点[J].药学学报,2004,39(5):328-332.
[27]董晞,赵世萍,刘岩,等.甘草苷对乌头碱致心肌细胞损伤的保护作用[J].中华中医药杂志,2009,24(2):163-165.
[28]付敏.乌头碱致心律失常的细胞分子机制的实验研究[D].北京:北京中医药大学,2007.
[29]张雪,赵炳祥,董艳红,等.人参皂苷Rb1配伍乌头碱对新生大鼠心肌细胞ATP酶及相关离子的影响[J].中国实验方剂学杂志,2016,22(7):112-115.
[2]杨宝峰,蔡本志.心律失常发病机制研究进展[J].国际药学研究杂志,2010,37(2):81-88.
[3]范新荣.心肌细胞内高钙引发心室肌电扰乱及心律失常[D].武汉:武汉科技大学,2009.
[4]曾秋棠,黄恺,祝武强,等.颅痛定对单个豚鼠心室肌细胞缺氧后复氧早期L-Ca电流的影响[J].临床心血管病杂志,2001,17(5):215-217.
[5]Pogwizd S M,Mc Kenzie J P,Cain M E.Mechanism underlying spontaneous and in induced ventricular arrhythmias in patients with idiopathic dilated cardiomyopthy[J].Circulation,1998,98(4):2404-2414.
[6]刘丽平,杨琳.心力衰竭的钙循环与室性心律失常的研究进展[J].世界核心医学期刊文摘:心脏病学,2005,1(5):13-16.
[7]魏执真,曹若楠,吴江丽,等.调脉汤治疗快速性心律失常的研究[J].中国医药学报,1992,7(3):14-16.
[8]解欣然,易京红,王军,等.调脉饮拆方抗心律失常作用的研究[J].中国实验方剂学杂志,2011,17(2):109-112.
[9]李红,牛欣,李国彰,等.调脉饮注射液抗心律失常的实验研究[J].中国中药杂志,2006,31(9):759-762.
[10]魏执真,谢欣然,李景,等.调脉饮及拆方对实验性室性心律失常的作用和分子机制研究[J].中华中医药学刊,2017,35(12):2983-2986.
[11]陈江斌,唐其柱,黄从新,等.丹皮酚对心肌细胞自律性和延迟后除极的影响[J].中国应用生理学杂志,1999,15(4)332-334.
[12]王腾,唐其柱,江洪.丹皮酚对豚鼠心肌细胞动作电位及钙通道电流的影响[J].武汉大学学报:医学版,2001,22(4):331-333.
[13]张金花,熊永爱.丹皮酚对大鼠缺血性心律失常及其心肌细胞miRNA-1表达的影响[J].中国实验方剂学杂志,2015,21(5):129-132.
[14]Ohta H,Matsumoto K,Shimizu M,et al.Paeoniflorin attenuates learning impairment of aged rats in operant brightness discrimination task[J].Pharmacol Biochem Behav,1994,49(1):213-217.
[15]张林,王停,杜守颖,等.芍药苷、薄荷脑对葛根素在Calu-3细胞模型转运过程中膜流动性、钠-钾离子ATP酶和钙离子ATP酶的影响[J].中国中药杂志,2018,43(4):731-735.
[16]刘和兰,莫丹,梁荣寿,等.芍药苷对异丙肾上腺素诱导大鼠心肌肥大的抑制作用及其机制研究[J].中国实验方剂学杂志,2015,21(16):88-92.
[17]张广钦,郝雪梅,陈世忠,等.芍药苷对大鼠心肌细胞L钙通道的阻断作用[J].中国药理学通报,2003,19(8):863-866.
[18]刘丹,曹广尚,司席席,等.黄连中生物碱类成分抗心律失常研究概述[J].山东中医杂志,2017,36(2):164-171.
[19]徐尚忠,张翼,任建英,等.小檗碱对豚鼠心室肌细胞L-及T-型钙离子通道的影响[J].中国药理学报,1997,18(6):515-518.
[20]司晓晨,R.Clintton Webb.黄连素片对钙离子通道的拮抗作用[J].南京中医药大学学报,1996,12(6):20-22.
[21]魏婷,梁喆,金彦,等.小檗碱、莲心碱和甲基莲心碱对HERG通道表达的影响[J].中国中药杂志,2013,38(2):239-244.
[22]Furukawa Y,XUE Y X,Chiba S,et al.Effects of zatebradine on ouabain,two-stage coronary ligation and epinephrine-induced ventricular tachyarrhythmias[J].Eur J Pharmacol,1996,300(3):203-210.
[23]Faber E M,Rudy Y.Action potential and contraetility changes in[Na+]ioverloaded cardiac myocytes:a simulation study[J].Bio Phys J,2000,78:2392-2404.
[24]Friese J,Gleitz J,Gutser U T,et al.Aconitum sp.alkaloids:the modulation of voltage-dependent Na+channels,toxieity and antinociceptive properties[J].Eur J Pharmaeol,1997,337(2/3):165-174.
[25]刘影,单宏丽,孙宏丽,等.乌头碱对大鼠心室肌细胞包浆钙及L-型钙电流的影响[C]∥中国病理生理学会动脉粥样硬化专业委员会.中国病理生理学会动脉粥样硬化专业委员会五届一次会议论文集:2002卷.衡阳:中国动脉硬化杂志,2002:101-102.
[26]龚冬梅,单宏丽,周宇宏,等.哇巴因和乌头碱诱发豚鼠和大鼠心律失常的离子作用靶点[J].药学学报,2004,39(5):328-332.
[27]董晞,赵世萍,刘岩,等.甘草苷对乌头碱致心肌细胞损伤的保护作用[J].中华中医药杂志,2009,24(2):163-165.
[28]付敏.乌头碱致心律失常的细胞分子机制的实验研究[D].北京:北京中医药大学,2007.
[29]张雪,赵炳祥,董艳红,等.人参皂苷Rb1配伍乌头碱对新生大鼠心肌细胞ATP酶及相关离子的影响[J].中国实验方剂学杂志,2016,22(7):112-115.