基于钙信号转导的稳心颗粒抗心律失常机制研究  

Research on antiarrhythmic mechanism of Wenxin Granules based on calcium signal transduction

作　　者：钱舒乐 赵虎成 于露 李晓凤[4] 郭海珍 赵玉珂 王润英 丛紫东[4] 杜武勋[4] QIAN Shule;ZHAO Hucheng;YU Lu;LI Xiaofeng;GUO Haizhen;ZHAO Yuke;WANG Runying;CONG Zidong;DU Wuxun(College of Traditional Chinese Medicine,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;Institute of Biomechanics and Medical Engineering,Department of Engineering Mechanics,School of Aerospace Engineering,Tsinghua University,Beijing 100084,China;Affiliated Hospital of Xuzhou Medical University,Xuzhou 221132,China;Second Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300250,China)

机构地区：[1]天津中医药大学中医学院,天津301617 [2]清华大学航天航空学院工程力学系生物力学与医学工程研究所,北京100084 [3]徐州医科大学附属医院,江苏徐州221132 [4]天津中医药大学第二附属医院,天津300250

出　　处：《中草药》2025年第3期855-866,共12页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金项目(82004329);天津市教委科研计划项目(2024KJ020);天津中医药大学新教师科研启动项目“雏鹰计划”项目(KG240215);国家中医药管理局全国名老中医药专家传承工作室建设项目“杜武勋全国名老中医药专家传承工作室”(国中医药人教函[2022]75号);天津市中医药管理局天津市名中医传承工作建设项目“杜武勋天津市名中医传承工作室”。

摘　　要：目的基于钙信号转导探讨稳心颗粒抗心律失常的可能作用机制。方法60只雄性SD大鼠随机分为对照组、模型组及稳心颗粒低、中、高剂量(1.34、2.68、5.36 g/kg)组和胺碘酮(35.7 mg/kg)组。各组大鼠连续预防性ig给药21 d,对照组和模型组ig纯水,末次给药1 h后麻醉大鼠,尾iv乌头碱以诱导心律失常。采用RM6240多道生理信号采集处理系统记录大鼠心电表现,并进行心电参数分析。采用蛋白质免疫印迹法(Western blotting,WB)检测心肌组织钙/钙调蛋白依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinaseⅡ,Ca MKⅡ)、兰尼碱受体2(ryanodine receptor 2,RyR2)和L-型钙通道(L-type calcium channel,LTCC)的蛋白表达。采用瞬时转染方法将编码LTCC(α1dδ、α2δ、β2α)和绿色荧光蛋白(green fluorescent protein,GFP)的质粒共转染至人胚胎肾细胞HEK293上,予以稳心颗粒含药培养液干预,用钙离子荧光探针(Fluo-4 AM)检测细胞内钙离子浓度的变化、膜片钳技术检测细胞膜上LTCC电流变化。结果尾iv乌头碱后大鼠出现心律失常,与对照组比较,模型组大鼠心率显著加快(P<0.05),心律失常评分显著升高(P<0.01),Ca MKⅡ和RYR2蛋白表达增加(P<0.01),LTCC蛋白表达减少(P<0.01);与模型组比较,稳心颗粒高剂量组和胺碘酮组心率显著减慢(P<0.05),心律失常评分降低(P<0.01),Ca MKⅡ和RYR2蛋白表达减少(P<0.05、0.01),LTCC蛋白表达增加(P<0.05、0.01)。稳心颗粒在0.78~12.50 g/L对细胞的促增殖作用最强(P<0.05、0.01);稳心颗粒能增加LTCC电流峰值和电流密度,导致电压依赖性通道激活曲线发生负向偏移、电压依赖性通道失活曲线发生正向偏移。结论稳心颗粒可能通过影响钙信号转导相关蛋白表达并改变L-型钙通道的门控动力学发挥抗心律失常效应。Objective To investigate the possible mechanism of anti-arrhythmic action of Wenxin Granules(稳心颗粒)based on calcium signal transduction.Methods In animal experiment,60 male SD rats were randomly divided into control group,model group,Wenxin Granules low,medium and high dose(1.34,2.68,5.36 g/kg)groups and amiodarone(35.7 mg/kg)group.The rats in each group were given continuous prophylactic administration for 21 d,and the control group and model group were given pure water.The rats were anesthetized 1 h after the last administration and injected with aconitine through the tail vein to induce arrhythmia.The electrocardiograph(ECG)performance of rats was recorded by RM6240 physiological signal acquisition and processing system,and the ECG parameters were analyzed.Western blotting(WB)was used to detect calcium/calmodulin-dependent protein kinase II(CaMKII),ryanodine receptor 2(RyR2)and L-type calcium channel(LTCC)protein expression in myocardial tissue.In the cell experiment,the plasmid encoding LTCC(α1δ,α2δ,β2α)and green fluorescent protein(GFP)was transfected into HEK293 cells by transient transfection method and interfered with Wenxin Granules,the change of intracellular calcium ion concentration was detected by calcium ion fluorescence probe(Fluo-4 AM)and the change of LTCC current on cell membrane was detected by patch clamp technique.Results:Arrhythmia occurred in rats after being injected with aconitine through the tail vein.Compared with the control group,the heart rate of the model group was faster(P<0.05),the arrhythmia score was higher(P<0.01),the expression of CaMKⅡand RYR2 protein was increased(P<0.01),and the expression of LTCC protein was decreased(P<0.01).Compared with model group,the heart rate of Wenxin Granules high-dose group and amiodarone group significantly slowed down(P<0.05),arrhythmia score was decreased(P<0.01),protein expressions of CaMKⅡand RYR2 decreased(P<0.05,0.01),and protein expression of LTCC increased(P<0.05,0.01).At the concentration of 0.78—12.50 g/L,Wenxin Gran

关 键 词：钙信号转导 心律失常 膜片钳 稳心颗粒 离子通道 三七皂苷R_(1) 人参皂苷Rg_(1) 人参皂苷Rb_(1) 

分 类 号：R285.5[医药卫生—中药学]