基于斑马鱼模型的何首乌水提物及其主要成分的肝毒性研究

Study on Hepatotoxicity of Aqueous Extract of Polygoni Multiflori Radix and Its Main Components Based on Zebrafish

目的研究何首乌水提物及主要的单体成分对斑马鱼幼鱼的肝毒性。方法选用72 hpf(hours post fertilization)的肝脏转基因斑马鱼幼鱼(lfabp:ds Red)为实验动物,以对乙酰氨基酚(APAP)为阳性药,同时给药不同浓度的何首乌水提物及主要单体成分2 d后,从体视显微镜下对斑马鱼幼鱼生存情况、肝脏形态进行观察,计算肝脏及卵黄囊的尺寸比,荧光显微镜下观察斑马鱼幼鱼肝脏荧光发光情况,并通过与对照组比对判断各成分肝毒性情况。结果 2 mg·mL^(-1)的何首乌水提物、2μg·mL^(-1)大黄素给药1 d后,斑马鱼全部死亡,20μg·mL^(-1)的大黄素-8-O-β-D-葡萄糖苷给药2 d后,斑马鱼全部死亡,其他单体成分及上述成分剩余浓度组斑马鱼未出现死亡;8 mM APAP组及10、20μg·mL^(-1)芦荟大黄素肝脏颜色变暗,且与对照组相比,肝脏尺寸明显减小(P<0.01),卵黄囊保留尺寸明显增加(P<0.01),肝脏的荧光表达减弱。其余各组及1μg·mL^(-1)芦荟大黄素组未见明显影响。结论 10、20μg·mL^(-1)芦荟大黄素对斑马鱼幼鱼具有肝脏毒性,何首乌水提物及其他单体成分在致死剂量下未表现出明显的致肝脏损伤作用。

Objective To investigate the hepatotoxicity of aqueous extract of Polygoni Multiflori Radix(AEPMR)and major monomer components in it with larvae zebrafish.Methods 72 hpf liver fluorescence transgenic larvae zebrafish(lfabp:dsRed)were treated with different doses of AEPMR and its main monomer components for 2 d respectively with acetaminophen(APAP)as positive drug.Larval zebrafish survival situation,liver morphology,change in size of liver and yolk sac retention,and the liver fluorescence are major factors to evaluate hepatotoxicity.Results Zebrafish all died at 2 mg·mL-1 AEPMR,2μg·mL-1 emodin(EM)after 1 d administration and all died at 20μg·mL-1 emodin-8-O-β-D-glucopyranoside(EG)after 2 d administration.There was no death in the rest doses and other monomer component groups.10μg·mL-1,20μg·mL-1 aloe-emodin(AE)and 8 mM APAP induced liver degeneration,reduced liver size(P<0.05 or P<0.01)and delayed yolk sac absorption(P<0.05 or P<0.01)in larval zebrafish.The liver fluorescence also decreased in larval zebrafish treated with 10μg·mL-1、20μg·mL-1 AE and 8 mmol·L-1 APAP,whereas AEPMR and other monomer component groups had no obvious effect on larval zebrafish liver.Conclusion 10μg·mL-1,20μg·mL-1 AE induced liver toxicity in larval zebrafish,AEPMR and other monomer component groups do not show obvious liver damage.