基于斑马鱼多模态评价杜仲配伍补骨脂对其香豆素成分的代谢及毒/效影响  

Evaluation of effects of Eucommiae Cortex combined with Psoraleae Fructus on metabolism and toxicity/efficacy of coumarin components based on zebrafish multimodal analysis

作　　者：高一波 李佳妍 石子琪 宁青[1,2,3] 韦英杰[1,2,3] GAO Yi-bo;LI Jia-yan;SHI Zi-qi;NING Qing;WEI Ying-jie(Affiliated Hospital of Integrated Traditional Chinese and Western Medicine,Nanjing University of Chinese Medicine,Nanjing 210028,China;Key Laboratory of New Drug Delivery Systems of Chinese Materia Medica,Jiangsu Province Academy of Traditional Chinese Medicine,Nanjing 210028,China;The Third Clinical Medical College,Nanjing University of Chinese Medicine,Nanjing 210028,China;Department of Pharmacy,Yantai Yuhuangding Hospital,Yantai 264000,China)

机构地区：[1]南京中医药大学附属中西医结合医院,江苏南京210028 [2]江苏省中医药研究院,国家中医药管理局中药释药系统重点研究室,江苏南京210028 [3]南京中医药大学第三临床医学院,江苏南京210028 [4]烟台毓璜顶医院药学部,山东烟台264000

出　　处：《中草药》2023年第5期1449-1460,共12页Chinese Traditional and Herbal Drugs

基　　金：国家“重大新药创制”科技重大专项(2017ZX09301056);国家中医药管理局2022年中央财政转移支付地方项目(2022GJJZDYJS-01);江苏省中医药科技发展计划项目(ZD202225);江苏省中医药科技发展计划项目(YB2020022);江苏省中医药科技发展计划项目(QN202214);江苏省医学创新团队项目(CXTDB2017003)。

摘　　要：目的用斑马鱼多模态评价杜仲配伍补骨脂对其香豆素糖苷的代谢及毒/效影响。方法用受精后1~6 d(day after fertilization,dpf)的斑马鱼评价杜仲提取物(lignan extract of Eucommiae Cortex,LEEC)配伍补骨脂香豆素提取物(coumarin extract of Psoraleae Fructus,CEPF)以及代表成分松脂醇二葡萄糖苷(pinoresinol diglucoside,PDG)配伍补骨脂苷(psoralenoside,PSS)和异补骨脂苷(isopsoralenoside,IPSS)的安全性,观察斑马鱼胚胎发育情况,包括胚胎致畸、致死情况,并计算斑马鱼半数死亡浓度(half lethal concentration,LC50);用1~6 dpf斑马鱼暴露于上述配伍组合物,分析PSS和/或IPSS及其代谢物补骨脂素(psoralen,PS)和/或异补骨脂素(isopsoralen,IPS)的动态变化;用25μmol/L泼尼松龙诱导斑马鱼骨质疏松模型,采用茜素红对培养至8 dpf的各给药组斑马鱼幼鱼骨骼染色,进行显微观察、数码成像,并用图像软件定量分析骨骼染色区域来评价上述配伍组合抗骨质疏松活性。结果LEEC配伍CEPF及PDG分别配伍PSS和IPSS,可改善斑马鱼心包、卵黄囊等形态畸变,减少鱼死亡率,提高鱼致畸或致死浓度;能明显减慢PSS和IPSS脱糖基转化为潜在肝损伤代谢产物PS和IPS的速率,减慢毒性代谢物的产生是杜仲配伍补骨脂减毒的重要因素;LEEC配伍CEPF以及PDG分别配伍PSS和IPSS可显著增加斑马鱼头骨累积光密度值,提示具一定增效作用。结论采用斑马鱼多模态高效评价了杜仲配伍对CEPF及补骨脂香豆素代表成分的代谢与毒/效作用的影响,为杜仲配伍补骨脂的减毒、增效作用研究提供高效的方法与思路。Objective To evaluate the effects of Duzhong(Eucommiae Cortex)combined with Buguzhi(Psoraleae Fructus)on its metabolism and toxicity/efficacy of coumarin components with zebrafish multimodal analysis.Methods Zebrafish 1-6 d after fertilization(1-6 dpf)was used to evaluate the safety of lignan extract of Eucommiae Cortex(LEEC)combined with coumarin extract of Psoraleae Fructus(CEPF)and the representative components of pinoresinol diglucoside(PDG)combined with psoralenoside(PSS)and isopsoralenoside(IPSS),zebrafish embryonic development including embryonic teratogenicity and lethality were observed,and half death concentration(LC50)of zebrafish was calculated;Zebrafish were exposed to the above compatible compositions at 1-6 dpf to analyze the dynamic changes of PSS and/or IPSS and their metabolites psoralen(PS)and/or isopsoralen(IPS);The zebrafish osteoporosis model was induced with 25μmol/L prednisolone,microscopic observation and digital imaging of zebrafish larvae of each treatment group cultured to 8 dpf were performed using alizarin red,and the bone staining area was quantitatively analyzed by image software to evaluate the anti-osteoporosis activity of the above compatibility combinations.Results LEEC combined with CEPF and PDG combined with PSS and IPSS could alleviate zebrafish morphology distortion of heart,yolk sac,etc.and reduce fish mortality,and increase teratogenic or lethal concentrations of fish,significantly slowed down the conversion rate of PSS and IPSS to potential liver injury metabolites PS and IPS,and slowed down the production of toxic metabolites was an important factor for the attenuating effect of Eucommiae Cortex combined with Psoraleae Fructus;LEEC combined with CEPF and PDG combined with PSS and IPSS could increase the cumulative optical density of zebrafish skull,suggesting a certain synergistic effect.Conclusion The effects of Eucommiae Cortex compatibility on metabolism and toxicity/efficacy of CEPF and its representative components were high efficiently evaluated by zebrafish mult

关 键 词：斑马鱼 补骨脂 杜仲 毒性 代谢 骨质疏松 补骨脂素 异补骨脂素 补骨脂苷 异补骨脂苷 松脂醇二葡萄糖苷 

分 类 号：R285.5[医药卫生—中药学]