[关键词]
[摘要]
目的 利用实时细胞分析(RTCA)Cardio系统,通过体外细胞实验及整体动物实验评价速效救心丸抗心律失常作用。方法 培养原代大鼠乳鼠心肌细胞,接种于E-Plate Cardio 96,分析3 600/孔、20 000/孔以及90 000/孔接种浓度心肌细胞的细胞指数(CI)和搏动图形;MTT法检测速效救心丸(10、20、40、80、160 μg/mL)对原代心肌细胞活力的影响;考察速效救心丸(160 μg/mL)对乌头碱(8 μmol/L)诱发心律失常的心肌细胞搏动频率、搏动幅度的影响;舌下iv乌头碱50 μg/kg制备大鼠心律失常模型,利用八导生理记录仪观察速效救心丸(54、270 mg/kg)对Ⅱ导联心电图的影响。结果 20 000/孔为最佳接种浓度,CI值在观察时间内成对数生长,细胞形成同步搏动,波形正常且持续时间长;在10~160μg/mL浓度范围内,速效救心丸对心肌细胞活力均没有明显影响。RTCA Cardio系统检测结果显示,与模型组比较,速效救心丸能显著降低心肌细胞搏动速率(P<0.01)、显著增加心肌细胞的搏动幅度(P<0.01)。动物实验发现,速效救心丸能有效对抗乌头碱诱导的心律失常、改善乌头碱对实验动物RR间期的抑制作用和对心率的促进作用、降低乌头碱诱导的室颤发生率(P<0.01、0.001)。结论 RTCA Cardio系统能实现对心肌细胞的实时监测,可以检测心肌细胞的搏动情况;速效救心丸能有效对抗乌头碱诱导的心律失常。
[Key word]
[Abstract]
Objective To evaluate the anti-arrhythmia effect of Suxiao Jiuxin Pill in vitro cell experimental and whole animal experiment by using the real-time cell analysis (RTCA) Cardio system, which provided experimental basis for the clinical application of Suxiao Jiuxin Pill. Methods The primary rat cardiac myocytes were cultured and inoculated in e-plate cardio 96. The cell index (CI) and pulsation pattern of cardiac myocytes at 3 600/well, 20 000/well and 90 000/well were analyzed. The effects of Suxiao Jiuxin Pill (10, 20, 40, 80, 160 μg/mL) on the viability of primary cardiac myocytes were detected by MTT method. The effects of Suxiao Jiuxin Pill (160 μg/mL) on aconitine (8 μmol/L) were investigated. The effects of tachycardia frequency and amplitude of cardiac myocytes on arrhythmia; the rat model of arrhythmia was made by sublingual aconitine 50 μg/kg, and the effects of Suxiao Jiuxin Pill (54, 270 mg/kg) on lead II ECG were observed by eight channel physiological recorder. Results In the concentration range of 10- 160 μg/mL, Suxiao Jiuxin Pill had no significant effect on the viability of cardiomyocytes. The results of RTCA cardio system showed that compared with the model group, Suxiao Jiuxin Pill could significantly reduce the beating rate of cardiomyocytes (P < 0.01), and significantly increase the beating range of cardiomyocytes (P < 0.01). Animal experiments showed that Suxiao Jiuxin Pillcan effectively resist aconitine induced arrhythmia, improve the inhibition of aconitine on RR interval and the promotion of heart rate, and reduce the incidence of ventricular fibrillation induced by aconitine (P < 0.01, 0.001). Conclusion RTCA cardio system can realize real-time monitoring of myocardial cells, Suxiao Jiuxin Pill can effectively treat arrhythmiawas induced by Aco.
[中图分类号]
R965
[基金项目]
国家自然科学基金资助项目(81703710);天津市留学回国人员科技活动启动项目(2018018)