斑马鱼幼鱼常山碱盐静脉注射给药和水溶液暴露给药毒性比较  

Comparison of toxicity of Dichroa alkali salt in zebrafish larvae via intravenous administration and aqueous solution exposure

作　　者：胡晓祯 马丽娜 郑长辉[1] 郭媛媛 叶祖光 曹俊岭[1,4] HU Xiaozhen;MA Lina;ZHENG Changhui;GUO Yuanyuan;YE Zuguang;CAO Junling(School of Chinese Materia Medica,Beijing University of Chinese Medicine Beijing 100029,China;Dongfang Hospital,Beijing University of Chinese Medicine,Beijing 100078,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Post-doctoral Scientific Research Center,Beijing 100700,China;Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China)

机构地区：[1]北京中医药大学中药学院,北京100029 [2]北京中医药大学东方医院药学部,北京100078 [3]中国中医科学院中药研究所,北京100700 [4]北京中医药大学东直门医院,北京100700

出　　处：《中国药理学与毒理学杂志》2024年第8期595-603,共9页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金(82104518)。

摘　　要：目的对比分析常山碱盐(DAS)iv给药和水溶液暴露给药毒性的区别。方法①将受精后2 d(2 dpf)发育良好的斑马鱼幼鱼随机分为正常对照组(无任何处理)、溶剂对照组(生理盐水,iv)和DAS组(0.125,0.25,0.50,1.00和2.00 mg·kg^(-1),iv),给药后连续观察3 d,以斑马鱼心率为0确定死亡,计算死亡率、最大非致死剂量(MNLD)和10%致死剂量(LD_(10))。给药后4 h,体视显微镜观察0.50和2.00 mg·kg^(-1)组斑马鱼静脉窦淤血、心包水肿及心率和血流变慢情况,并计算其发生率。斑马鱼iv给予DAS 0.041,0.136,0.412和0.452 mg·kg^(-1),给药后连续3 d体视显微镜观察并计算斑马鱼幼鱼发育畸形表型,包括心包水肿、心率异常、血流变慢、循环缺失、眼睛异常、脑部畸形、下颌异常、肝缺失/变性、卵黄囊吸收延迟、肠腔异常、身体着色异常、身体水肿、躯干/尾/脊索弯曲和肌肉变性等发生率。②斑马鱼幼鱼随机分为正常对照组和DAS水溶液暴露(2.5,5.0,10.0,25.0,50.0,75.0和100.0 mg·L^(-1))组,连续暴露并观察3 d,计算死亡率、LD10和MNLD。斑马鱼暴露于DAS 0.32,1.06,3.20和11.00 mg·L^(-1)水溶液,连续暴露3 d,体视显微镜观察斑马鱼幼鱼发育畸形表型,并计算其发生率。结果①斑马鱼幼鱼iv给予DAS的MNLD和LD10分别为0.412和0.452 mg·kg^(-1)。与溶剂对照组相比,iv给予DAS 4 h后0.50和2.00 mg·kg^(-1)组斑马鱼静脉窦淤血、心率减慢和心包水肿发生率均显著增加(P<0.05,P<0.01),2.00 mg·kg^(-1)组血流变慢发生率亦显著增加(P<0.01);DAS 0.041,0.136,0.412和0.452 mg·kg^(-1)组斑马鱼卵黄囊吸收延迟率显著增加(P<0.05,P<0.01),0.452 mg·kg^(-1)组斑马鱼死亡率显著增加(P<0.05),且死亡斑马鱼具有心包水肿现象。②斑马鱼DAS水溶液暴露的MNLD和LD10分别为3.20和11.00 mg·L^(-1)。与正常对照组相比,3.20和11.00 mg·L^(-1)组斑马鱼心率减慢和血流变慢发生率均明显增加(P<0.01),1.06,3.20和11.00 mg·L^(OBJECTIVE To study the differences in toxicity between intravenous(iv)administration and aqueous solution exposure of Dichroa alkali salt(DAS)in zebrafish.METHODS①Well-devel⁃oped zebrafish larvae of 2 d post fertilization(2 dpf)were randomly divided into the normal control(no treatment),solvent control(saline,iv),and DAS groups(0.125,0.25,0.50,1.00 and 2.00 mg·kg^(-1),iv)before being observed for 3 consecutive days after administration.A heart rate of 0 was determined as death of zebrafish,and the mortality rate,maximum non-lethal dose(MNLD),and 10 percent lethal dose(LD10)were calculated.The incidence of venous sinus congestion,pericardial edema,slowing heart rate and blood flow of zebrafish in the 0.50 and 2.00 mg·kg^(-1) groups were observed and calculated by somatoscopic microscopy at 4 h after drug administration.Zebrafish larvae were iv given DAS at doses of 0.041,0.136,0.412,and 0.452 mg·kg^(-1) while the malformation phenotypes of zebrafish larvae development were observed under a stereomicroscope for 3 consecutive days,including pericardial edema,abnormal heart rate,slow blood flow,loss of circulation,eye abnormalities,brain malforma⁃tions,jaw abnormalities,loss/degeneration of the liver,delayed yolk sac absorption,intestinal abnormal⁃ities,abnormal body coloration,body edema,curvature of the trunk/tail/nodal cord and muscle degener⁃ation before the incidence was calculated.②Zebrafish larvae were randomly divided into a normal control group and DAS aqueous solution exposure groups at concentrations of 2.5,5.0,10.0,25.0,50.0,75.0,and 100.0 mg·L^(-1),observed for 3 d until the mortality rate,LD10,and MNLD were calculated.Zebrafish were exposed to DAS aqueous solutions at concentrations of 0.32,1.06,3.20,and 11.00 mg·L^(-1),and the malformation phenotypes of zebrafish larvae development were observed under a stereomicro⁃scope for 3 consecutive days to calculate the incidence.RESULTS①The MNLD and LD_(10) of DAS iv administered to zebrafish larvae were 0.412 and 0.452 mg·kg^(-1),respecti

关 键 词：常山碱盐 斑马鱼 静脉注射 水溶液暴露 毒性评价 

分 类 号：R285.1[医药卫生—中药学]