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南京医科大学学报(自然科学版) 第43卷第12期
·1630 · Journal of Nanjing Medical University(Natural Sciences) 2023年12月
·基础研究·
氯氮平对斑马鱼心脏发育的影响及机制研究
黄凯玥 ,张 伟 ,朱玉婕 ,孙玉洁 1,2* ,袁 栎 2,3*
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南京医科大学细胞生物学系,江苏省人类功能基因组学重点实验室,生物化学与分子生物学系,江苏 南京 211166
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[摘 要] 目的:探索氯氮平(clozapine,CLP)对斑马鱼心脏发育的损伤效应及相关机制。方法:以 12.5 μmol/L 氯氮平(L⁃
CLP)及25.0 μmol/L氯氮平(H⁃CLP)处理斑马鱼胚胎72 h,显微镜下检测心脏形态、心率、静脉窦⁃动脉球(sinus venosus⁃bulbus
arteriosus,SV⁃BA)距离等指标;实时荧光定量聚合酶链反应(RT⁃PCR)检测心脏发育相关基因NKX2.5、Hand2及Cmlc2表达;此
外,H⁃CLP 组处理 24 h 后,利用内质网抑制剂 4⁃苯基丁酸(4⁃phenylbutyric acid,4⁃PBA)或多巴胺 D2 受体抑制剂溴隐亭(bro⁃
mocriptine,BRC)联合处理48 h后,统计心脏功能指标,检测内质网应激分子(Chop、Bip)及凋亡相关分子(Caspase3、Bax/Bcl2)
的表达。结果:CLP剂量依赖性导致斑马鱼心脏结构及功能损伤,Chop、Bip和Caspase3、Bax/Bcl2的表达增加;4⁃PBA及BRC显
著缓解CLP诱导的心脏损伤及Chop、Bip、Caspase3、Bax/Bcl2的高表达。结论:CLP导致斑马鱼心脏发育受损,可能与抑制多巴
胺D2受体、激发内质网应激有关。
[关键词] 氯氮平;心脏毒性;斑马鱼胚胎;内质网应激
[中图分类号] Q344 [文献标志码] A [文章编号] 1007⁃4368(2023)12⁃1630⁃06
doi:10.7655/NYDXBNS20231202
Study on the toxicity of clozapine to the heart development of zebrafish embryos
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HUANG Kaiyue ,ZHANG Wei ,ZHU Yujie ,SUN Yujie ,YUAN Li 2,3*
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1 Department of Cell Biology,Key Laboratory of Human Functional Genomics of Jiangsu Province,Department of
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Biochemistry and Molecular Biology,Nanjing Medical University,Nanjing 211126,China
[Abstract] Objective:To explore the potential damage effects and related mechanisms of clozapine(CLP)on the heart development
of zebrafish. Method:12.5 μmol/L CLP(L⁃CLP)and 25.0 μmol/L CLP(H⁃CLP)were used to culture zebrafish embryos for 72 hours,
respectively. The heart morphology,heart rate,and the distance between venous sinus and arterial bulb(SV⁃BA)were observed under
microscope;The expression of cardiac development⁃related genes NKX2.5,Hand2 and Cmlc2 were detected by fluorescence quantitative
PCR;In addition,after 24 hours of culture,the endoplasmic reticulum inhibitor 4⁃phenylbutyric acid(4⁃PBA)or dopamine D2 receptor
inhibitor bromocriptine(BRC)were added to the H ⁃ CLP group for 48 hours. Then,we counted the cardiac function indicators,and
detected the expression of endoplasmic reticulum stress genes(Chop、Bip)and apoptosis⁃related genes(Caspase3,Bax/Bcl2). Results:
CLP induced damages of cardiac structure and function in a dose⁃dependent manner,induced the expression of Chop and Bip,and further
increased the expression of Caspase3 and Bax/Bcl2. 4⁃PBA and BRC significantly alleviated CLP⁃induced cardiac injury and decreased
the expression levels of Chop、Bip、Caspase⁃3 and Bax/Bcl2. Conclusion:CLP can damage the heart development of zebrafish,and the
mechanism may be related to inhibiting dopamine D2 receptor and stimulating endoplasmic reticulum stress.
[Key words] clozapine;cardiac toxicity;zebrafish embryo;endoplasmic reticulum stress
[J Nanjing Med Univ,2023,43(12):1630⁃1635]
先天性心脏病(congenital heart disease,CHD)是 新生儿最常见的先天性异常疾病,并导致超过40%
[1]
的产前死亡 。我国每年新发 CHD 的总经济负担
[基金项目] 国家自然科学基金项目(81803644);江苏省高
超过 126 亿元,给患者家庭和社会带来巨大的医疗
等学校自然科学研究重大项目(17KJA310001)
[2]
通信作者(Corresponding author),E⁃mail:yuanli@njmu.edu.cn;
∗ 负担和精神负担 。目前研究表明 CHD 是胚胎期
yujiesun@njmu.edu.cn 遗传与环境因素共同作用所导致。研究发现,孕期