摘要
目的:建立大鼠灌胃乌头碱诱发心律失常伴心肌细胞凋亡模型,观察苦参总碱对模型大鼠的影响,探讨苦参总碱对乌头碱诱发心肌细胞凋亡的保护作用。方法:利用乌头碱对人源干细胞诱导的心肌细胞(human cardiomyocytes,hCM)进行刺激,实时监测其搏动节律,观察经过苦参总碱干预后心肌细胞搏动节律的变化。连续10 d灌胃给予大鼠乌头碱0.00146 g/kg(10 mL/kg),制备乌头碱诱发大鼠心律失常伴心肌细胞凋亡模型,各组同时分别给予苦参总碱0.08、0.04、0.02 g/kg(10 mL/kg)进行干预;12 d后将大鼠麻醉测量心电图改变,对心脏组织进行TUNEL染色观察细胞凋亡率,并检测钙离子释放通道ryanodines受体2蛋白(RyR2)含量以及肌浆网钙泵(SERCA)蛋白的表达,探讨苦参总碱对乌头碱致心肌细胞凋亡的保护作用及其作用机制。结果:与正常对照组比较,乌头碱组心肌搏动节律明显增加(P<0.05),与乌头碱组比较,苦参总碱50、100μg/mL组心肌细胞搏动节律明显降低(P<0.05)。在乌头碱灌胃诱发大鼠心律失常伴心肌细胞凋亡模型中,苦参总碱能够改善乌头碱导致的心律失常,与正常对照组比较,模型对照组的T波高度发生明显改变(P<0.05),与模型对照组相比,胺碘酮0.055 g/kg组、苦参总碱0.08、0.04 g/kg组能够明显抑制乌头碱引起的T波变化(P<0.05);与正常对照组比较,模型对照组心肌细胞凋亡率显著增加(P<0.01),心肌组织中Bcl-2蛋白表达显著下调(P<0.01),Bax、Caspase-3、Caspase-9表达明显上调(P<0.05或P<0.01),与模型对照组比较,苦参总碱0.08 g/kg组能够降低心肌细胞的凋亡率(P<0.05),心肌Bcl-2的蛋白表达明显上调(P<0.05),Bax、Caspase-9蛋白的表达明显下调(P<0.05);与正常对照组比较,模型对照组动物心肌组织中的RyR2含量明显升高(P<0.05),SERCA蛋白表达明显下调(P<0.05),与模型对照组比较,苦参总碱0.04、0.08 g/kg能够降低心肌组织中RyR2的含量(P<0.05),苦参总碱0.02、0.04、0.08 g/kg能上调动物心肌组织中SERCA蛋白的表达(P<0.05)。结论:苦参总碱能够显著抑制心肌细胞搏动节律增加,拮抗心律失常的发生及大鼠心电图T波变化,降低心肌细胞凋亡率。其对乌头碱致大鼠心律失常同时出现心肌细胞凋亡的保护作用机制可能是通过改善心肌细胞兴奋-收缩耦联中钙离子通道蛋白的表达、调节心肌细胞凋亡蛋白表达来实现的。
Objective:To observe the effect of matrine on the arrhythmia model with cardiomyocyte apoptosis in rats induced by oral administration of aconitine and explore its protective effect against aconitine-induced cardiomyocyte apoptosis.Methods:Human-induced pluripotent stem cell-derived cardiomyocytes(hCM)were stimulated by aconitine.The beating rhythm was monitored in real time and the changes in the beating rhythm of cardiomyocytes after matrine intervention were observed.The arrhythmia model with cardiomyocyte apoptosis was induced in rats by intragastrical administration of aconitine at 0.00146 g/kg(10 mL/kg)for 10 consecutive days.Subsequently,matrine was provided at 0.08 g/kg,0.04 g/kg,and 0.02 g/kg(10 mL/kg)for intervention at the same time.Twelve days later,the rats were anesthetized for electrocardiogram measurement.The heart tissues were stained with TUNEL to observe the cell apoptosis rate.The expression of the calcium release channel ryanodine receptor 2(RyR2)and sarcoplasmic reticulum calcium transport ATPase(SERCA)was measured to explore the protective effect and mechanism of matrine on aconitine-induced cardiomyocyte apoptosis.Results:Compared with the normal control group,the model group showed accelerated beating of cardiomyocytes(P<0.05),while compared with the model group,the matrine groups(50 and 100μg/mL)showed slowed beating(P<0.05).As revealed by the results,matrine could improve aconitine-induced arrhythmia based on the arrhythmia model with cardiomyocyte apoptosis in rats induced by intragastric administration of aconitine.Compared with the normal control group,the model group showed altered T wave height(P<0.05).Compared with the model group,the amiodarone group(0.055 g/kg)and the matrine groups(0.08 g/kg and 0.04 g/kg)showed inhibited T wave changes caused by aconitine(P<0.05).Compared with the normal control group,the model group showed increased apoptosis rate of cardiomyocytes(P<0.01),down regulated protein expression of Bcl-2 in myocardial tissues(P<0.01),and up regulated expression of Bax,Caspase-3,and Caspase-9(P<0.05 or P<0.01).Compared with the model group,the matrine group(0.08 g/kg)showed reduced apoptosis rate of cardiomyocytes(P<0.05),up regulated protein expression of Bcl-2(P<0.05),and down regulated protein expression of Bax and Caspase-9(P<0.05).Compared with the normal control group,the model group showed increased content of RyR2 protein in the myocardial tissues(P<0.05)and decreased protein expression of SERCA(P<0.05).Compared with the conditions in the model group,matrine at 0.08 g/kg and 0.04 g/kg could reduce the content of RyR2 in myocardial tissues(P<0.05),while that at 0.08 g/kg,0.04 g/kg,and 0.02 g/kg could up regulate the protein expression of SERCA in myocardial tissues(P<0.05).Conclusion:Matrine can inhibit the accelerated beating of cardiomyocytes,antagonize arrhythmia,inhibit T wave changes,and reduce the apoptosis rate of cardiomyocytes.Its mechanism against aconitine-induced arrhythmias with cardiomyocyte apoptosis may be related to the improvement of the expression of calcium channel proteins in the excitation-contraction coupling of cardiomyocytes and the regulation of the expression of cardiomyocyte apoptosis-related proteins.
作者
林玥
丁涛
陈革
王鑫
温富春
纪凤兰
刘博
徐惠波
Lin Yue;Ding Tao;Chen Ge;Wang Xin;Wen Fuchun;Ji Fenglan;Liu Bo;Xu Huibo(Changchun University of Chinese Medicine,Changchun 130117;Chinese Medicine Sciences Academy of Jilin Province,Changchun 130012;Changchun People Pharmaceutical Group Co.,Ltd.,Changchun 130031)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2021年第6期32-37,共6页
Pharmacology and Clinics of Chinese Materia Medica
基金
吉林省科技发展计划项目(编号:20140520040JH)
吉林省科技厅项目(编号:20200403128SF)
作者简介
通信作者:徐惠波,研究员,主要从事中药基础及应用开发研究,E-mail:xhb_6505@163.com;林玥,硕士在读,主要从事中药药理学研究,E-mail:1021401347@qq.com。
