氯氮平通过诱导斑马鱼胚胎心脏细胞凋亡引起心脏毒性  

Clozapine induced cardiotoxicity in zebrafish embryos through apoptosis

作　　者：张凤[1,2] 韩利文 张云 韩建 侯海荣[2] 王希敏 刘可春 田青平 何秋霞[2] ZHANG Feng;HAN Li-wen;ZHANG Yun;HAN Jian;HOU Hai-rong;WANG Xi-min;LIU Ke-chun;TIAN Qing-ping;HE Qiu-xia(School of Pharmacy,Shanxi Medical University,Taiyuan 030001,China;Biology Institute,Qilu University of Technology（Shandong Academy of Sciences）,Shandong Provincial EngineeringLaboratory for Biological Testing Technology,Key Laboratory for Biosensor of Shandong Province,Key Laboratory for Drug Screening Technology of Shandong Academy of Sciences,Jinan 250014,China)

机构地区：[1]山西医科大学药学院,山西太原030001 [2]齐鲁工业大学(山东省科学院),山东省科学院生物研究所,山东省生物检测技术工程实验室,山东省生物传感器重点实验室,山东省科学院药物筛选重点实验室,山东济南250014

出　　处：《中国药理学通报》2018年第8期1088-1093,共6页Chinese Pharmacological Bulletin

摘　　要：目的观察氯氮平对斑马鱼胚胎心脏的毒性,研究其毒性作用的机制。方法以24 hpf(hours post fertilization)斑马鱼胚胎为研究模型,以不同浓度的氯氮平(12.5、25、37.5、50、62.5μmol·L^(-1))进行处理,于处理24、48、72 h后观察斑马鱼的死亡率、心率和心脏形态变化。采用吖啶橙染色初步考察氯氮平产生心脏毒性的作用机制;利用实时定量PCR检测与凋亡相关的基因Bcl-2、Bax、caspase-9、caspase-3的表达。结果氯氮平可引起斑马鱼出现体长明显缩短、心包水肿、心脏变小、心率下降等现象,这些具有剂量依赖性。吖啶橙染色结果显示,氯氮平可剂量依赖性地诱导斑马鱼心脏细胞发生凋亡。给药处理72 h后,斑马鱼胚胎细胞抗凋亡基因Bcl-2的表达水平降低,促凋亡基因Bax的表达明显升高,Bcl-2/Bax表达比值明显下降;caspase-9和caspase-3表达均明显增加。结论氯氮平可导致斑马鱼胚胎产生心脏毒性,其毒性作用可能与其诱导胚胎细胞发生凋亡有关。Aim To observe the toxic effects of clozapine on zebrafish embryos and investigate the possible mechanisms underlying its cardiac toxicity. Methods Zebrafish of 24 hpf（hours post fertilization） were exposed to different concentrations of clozapine. After treatment for 24,48 and 72 h,the heart rates of the zebrafish embryos were examined and morphological changes of the heart were determined. The mechanism of clozapine induced cardiac toxicity was preliminarily explored by means of acridine orange（AO） staining.The mRNA levels of Bcl-2,Bax,caspase-9,caspase-3 were detected by real-time PCR analysis. Results Clozapine induced the decrease of body length,formation of pericardial edema,and decrease of heart rates of the embryos in a time and dose-dependent manner.AO staining showed that clozapine induced cell apoptosis of the cardiomyocytes. Real-time PCR analysis showed that the anti-apoptotic gene Bcl-2 was downregulated and pro-apoptosis gene Bax was up-regulated at 72 h post treatment,resulting in the decreased expression ratio of Bcl-2/Bax. In addition, caspase-9 and caspase-3 were induced significantly by clozapine.Conclusion Clozapine caused cardiac toxicity in zebrafish embryos by inducing cell apoptosis.

关 键 词：氯氮平 心脏毒性 斑马鱼胚胎 细胞凋亡 BCL-2/BAX caspase 

分 类 号：R-332[医药卫生] R322.11