北细辛水提物对斑马鱼胚胎发育、心脏和肝脏的毒性作用

doi:10.13481/j.1671-587X.20260510

摘要/Abstract

摘要：

目的基于斑马鱼胚胎模型系统评价北细辛水提物的发育毒性及潜在的心脏和肝脏相关毒性，明确其剂量-反应关系及敏感靶器官，为北细辛的安全性评价提供实验依据。方法选取受精后天数（dpf）为1的野生型AB品系斑马鱼胚胎，剔除未受精及发育异常个体后，随机分为对照组（仅加入培养液）及不同浓度北细辛水提物（100、200、300、500、1 000和2 000 mg·L^-1）处理组，每组18枚，持续暴露至5 dpf。记录各组斑马鱼胚胎2 dpf孵化率和5 dpf死亡率，绘制浓度-效应曲线，采用Probit回归分析计算半数致死浓度（LC₅₀）和20%致死浓度（LC₂₀）、10%致死浓度（LC₁₀）及5%致死浓度（LC₅）。于5 dpf观察各组斑马鱼胚胎形态学改变，测定各组斑马鱼心率、静脉窦-动脉球（SV-BA）距离和心包面积以评价胚胎心脏结构与功能变化。采用油红O染色测定各组斑马鱼胚胎肝脏积分光密度（IOD）以评价肝脏脂质沉积水平。结果与对照组比较，≥300 mg·L^-1组斑马鱼胚胎孵化率均明显降低（P<0.05），死亡率均明显升高（P<0.01），呈浓度依赖性。Probit回归分析，北细辛水提物LC₅₀为641.11 mg·L^-1，LC₂₀、LC₁₀和LC₅分别为317.78、223.33及146.67 mg·L^-1。形态学观察，≥200 mg·L^-1组斑马鱼胚胎可见卵黄囊吸收延迟、心包水肿、下颌畸形及体长缩短等异常表现。与对照组比较，LC₁₀和LC₂₀组斑马鱼胚胎心率明显降低（P<0.01），SV-BA距离和心包面积明显增加（P<0.01），提示斑马鱼胚胎心脏结构发育和功能状态受到影响。油红O染色，LC₅、LC₁₀和LC₂₀组斑马鱼胚胎均可见肝脏区域脂质沉积增加，肝脏脂质沉积水平值较对照组均明显升高（P<0.01），其中LC₂₀组胚胎肝脏脂质沉积程度最高。结论北细辛水提物对斑马鱼胚胎具有浓度依赖性的发育毒性，并可在低于LC₅₀的水平下诱发心脏结构和功能异常及肝脏中性脂质沉积增加，提示心脏和肝脏可能是其敏感靶器官。

关键词: 北细辛, 斑马鱼胚胎, 发育毒性, 心脏毒性, 肝毒性

Abstract:

Objective To systematically evaluate the developmental toxicity and potential cardiac and hepatic toxicity of the aqueous extract of Asarum heterotropoides based on zebrafish embryo model， to clarify its dose-response relationship and sensitive target organs， and to provide experimental evidence for the safety assessment of Asarum heterotropoides. Methods Wild-type AB strain zebrafish embryos at 1 day post-fertilization （dpf） were selected. After excluding unfertilized and abnormally developed individuals， they were randomly divided into a control group （culture medium only） and groups treated with different concentrations of the aqueous extract of Asarum heterotropoides （100， 200， 300， 500， 1 000， and 2 000 mg·L?1）， with 18 embryos in each group. Exposure lasted until 5 dpf. The hatching rate at 2 dpf and mortality rate at 5 dpf of zebrafish embryos in various groups were recorded. Concentration-effect curves were plotted， and Probit regression analysis was performed to calculate the median lethal concentration （LC₅₀）， as well as 20% lethal concentration （LC₂₀）， 10% lethal concentration（LC₁₀）， and 5% lethal concentration（LC₅）. At 5 dpf， morphological changes in zebrafish embryos in various groups were observed. Heart rate， sinus venosus-bulbus arteriosus （SV-BA） distance， and pericardial area of zebrafish embryos in various groups were measured to evaluate embryonic cardiac structure and function. Oil Red O staining was used to determine the hepatic integrated optical density （IOD） values to assess the levels of lipid accumulation in the liver of zebrafish embryos in various groups. Results Compared with control group， the hatching rates of zebrafish embryos in 300 mg·L^-1 group and higher concentration groups were significantly decreased （P<0.05）， while the mortality rates were significantly increased （P<0.01）， in a concentration-dependent manner. Probit regression analysis results showed that the LC₅₀ of the aqueous extract of Asarum heterotropoides was 641.11 mg·L^-1， and the LC₂₀， LC₁₀， and LC₅ were 317.78， 223.33， and 146.67 mg·L^-1， respectively. Morphological observation revealed that the zebrafish embryos in 200 mg·L^-1 group and higher concentration groups exhibited abnormalities such as delayed yolk sac absorption， pericardial edema， jaw malformation， and shortened body length. Compared with control group， the heart rates of zebrafish embryos in LC₁₀ group and LC₂₀ group were significantly decreased （P<0.01）， while the SV-BA distances and pericardial areas of zebrafish embryos were significantly increased （P<0.01）， indicating impaired cardiac structural development and functional status. The Oil Red O staining results showed increased lipid deposition in the liver of the zebrafish embryos in LC₅ group， LC₁₀ group and LC₂₀ group， and the levels of lipid accumulation in the liver were significantly higher than that in control group （P<0.01）， among which LC₂₀ group showed the highest degree of lipid deposition in the liver. Conclusion The aqueous extract of Asarum heterotropoides showed concentration-dependent developmental toxicity on zebrafish embryos and may induce cardiac structural and functional abnormalities as well as increased hepatic neutral lipid deposition at doses below the LC₅₀. The results suggested that the heart and liver may be its sensitive target organs.

Key words: Asarum heterotropoides, Zebrafish embryo, Developmental toxicity, Cardiotoxicity, Hepatotoxicity

中图分类号:

R285.5

郝博宇,李欣桦,王潇,汤雯翀,赵宏,张伟东. 北细辛水提物对斑马鱼胚胎发育、心脏和肝脏的毒性作用[J]. 吉林大学学报(医学版), 2026, 52(5): 1282-1290.

Boyu HAO,Xinhua LI,Xiao WANG,Wenchong TANG,Hong ZHAO,Weidong ZHANG. Toxic effects of Asarum heterotropoides aqueous extract on development, heart, and liver in zebrafish embryos[J]. Journal of Jilin University(Medicine Edition), 2026, 52(5): 1282-1290.

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