吴茱萸碱对心脏毒性的体内和体外实验研究

In vitro and in vivo Evaluation of the Cardiotoxicity of Evodiamine

目的:通过体内和体外实验探讨吴茱萸碱对心脏的毒性。方法:原代培养SD乳鼠心肌细胞,采用CCK-8检测试剂盒观察不同浓度吴茱萸碱对大鼠心肌细胞生存活力影响,同时观察不同浓度吴茱萸碱对心肌细胞乳酸脱氢酶(LDH)释放量、丙二醛(MDA)水平和超氧化物歧化酶(SOD)活性的影响;利用斑马鱼在体实验,测定吴茱萸碱最大非致死浓度(MNLC)和10%致死浓度(LC10),并观察吴茱萸碱对斑马鱼心率、心律和心脏形态学改变。结果:31.3μg·ml-1浓度及以上的吴茱萸碱可降低心肌细胞生存活力,增加LDH释放量和MDA水平,降低SOD活性。吴茱萸碱对斑马鱼的MNLC为113.4 ng·ml-1,LC10为354 ng·ml-1。与对照组相比,吴茱萸碱可导致斑马鱼心率降低,心律无明显变化,心包形态学改变和静脉窦和动脉球直线距离增大。结论:通过在体和离体试验发现吴茱萸碱对心脏具有一定的毒性,可能与氧化应激有关。

Objective: To explore the toxic effects of evodiaine on heart through in vivo and in vitro experiments. Methods: Primary neonatal rat cardiomyocytes were cultured,and the effects of evodiamine on the cell viability were assessed using CCK-8 assay. The effects of different concentrations of evodialine on the release levels of LDH,and MDA and SOD activity of cardiomyocytes were observed. The maximum non-lethal concentration(MNLC) and 10% lethal concentration(LC10) of evodiamine were determined using zebra fish in vivo experiment. The effects of evodialine on the heart rate,heart rhythm and heart morphology of zebra fish were observed.Results: Evodiline at the concentration ≥31.3 μg·ml-1 could reduce the viability of cardiomyocytes,increase LDH release and MDA level,and reduce SOD activity. The MNLC of evodiaine to zebra fish was 113.4 ng·ml-1,and the LC10 was 354 ng·ml-1. Compared with the control group,evodiamine could decrease the heart rate of zebra fish and showed no obvious effect on the heart rhythm,meanwhile,it could change the pericardial morphology and increase the linear distance between the venous sinus and the arterial bulb. Conclusion: Through in vitro and in vivo tests,it has been found that evodialine has a certain toxic effect on heart,which may be related to oxidative stress.