芫花醇提物对不同种属体系UGTs及UGT1A1活性影响.docVIP

芫花醇提物对不同种属体系UGTs及UGT1A1活性影响 　　摘要：目的 考察芫花醇提物对UGTs及UGT1A1活性的影响，为阐释芫花毒性机制提供依据。方法 采用体外肝微粒体孵育模型，以4-硝基酚为底物检测UGTs活性，以β-雌二醇为底物检测UGT1A1活性，利用UV和HPLC测定底物及代谢物含量。结果 芫花醇提物中3种黄酮类成分芹菜素、羟基芫花素、芫花素含量分别为6.34%、8.72%、6.06%，UV测得总二萜含量为31.40%。体外实验表明，在大鼠肝微粒体（RLM）和人肝微粒（HLM）孵育体系中，芫花醇提物均能显著抑制UGTs活性；对UGT1A1活性，在RLM、HLM和重组酶（rhUGT1A1）孵育体系中，芫花醇提物均表现为中等强度的抑制作用，以羟基芫花素计，半数抑制浓度分别为46.32、32.49、8.382 ?mol/L，抑制类型分别为竞争性抑制作用、反竞争性抑制作用和竞争性抑制作用。结论 芫花醇提物对不同肝微粒体孵育体系中UGTs及UGT1A1活性均可产生抑制作用且存在种属差异性，这种抑制作用可能是芫花致肝损伤的机制之一。 　　关键词：芫花；醇提物；UGTs；UGT1A1；肝微粒体；毒性；抑制作用 　　DOI：10.3969/j.issn.1005-5304.2017.04.021 　　中图分?号：R285.5 文献标识码：A 文章编号：1005-5304（2017）04-0083-05 　　Effects of Genkwa Flos Ethanol Extracts on UGTs and UGT1A1 Activities in Different Systems GUO Chang-e， MIAO Pei-pei， CHEN Hong-ying， CHEN Ning， MA Peng-kai， LI Hong-pin， ZHU Hong-yu， GAO Xing， ZHANG Yu-jie （School of Chinese Materia Medica， Beijing University of Chinese Medicine， Beijing 100102， China） 　　Abstract： Objective To investigate the inhibition of Genkwa Flos ethanol extracts on UGTs and UGT1A1 activities of different liver microsomes； To predict the toxicity mechanism of Genkwa Flos. Methods By adopting in vitro liver microsomes incubation model， 4-nitrophenol （4-NP） and β-estradiol were selected as substrates to determine activities of UGTs and UGT1A1 by UV and HPLC， respectively. Results Content determination results show that there were 6.34% of apigenin， 8.72% of hydroxygenkwanin and 6.06% of genkwanin in ethanol extracts， and total diterpene accounted for 31.40%. In vitro research showed that ethanol extracts significantly inhibited UGTs activity in rat and human liver microsome. UGT1A1 activity was inhibited by hydroxygenkwanin with IC50 about 46.32， 32.49 and 8.382 ?mol/L in rat liver microsome （RLM）， human liver microsome （HLM） and recombinant UGT1A1 （rhUGT1A1）， respectively. The inhibition types were competitive inhibition in RLM and rhUGT1A1， and uncompetitive inhibition in HLM. Conclusion Ethanol extracts showed different inhibition of UG