目的 以胆红素代谢过程中UGT1A1酶介导的胆红素葡糖醛酸结合环节为切入点,考察何首乌提取物体内、外肝毒性。方法 以胆红素为UGT1A1酶底物,考察给药后大鼠肝微粒体体系中UGT1A1酶活性的变化(体内),以及何首乌提取物在人肝微粒体、大鼠肝微粒体,人重组UGT1A1酶中(体外)对胆红素葡糖醛酸结合的抑制作用,以表观抑制常数Ki及Km,Vmax变化为评价指标,预测其肝毒性有无及大小。结果 何首乌提取物在三个体外体系中,对UGT1A1酶均有较强抑制作用,且抑制类型均为竞争型抑制。大鼠体内实验结果显示,何首乌提取物对UGT1A1酶有强抑制作用,抑制类型为反竞争型抑制。结论 本实验所建立的体外研究方法为中药肝毒性药物的筛选提供了新思路和新方法,对中药安全性评价具有借鉴意义。
Abstract
OBJECTIVE To study the hepatotoxicity of Polygonum multiflorum on the basic of the bilirubin metabolism mediated by glucuronidation of UGT1A1 enzyme. METHODS Inspected the enzyme kinetic parameters after giving the rats Polygonum multiflorum extract orally(in vivo), and added the Polygonum multiflorum extract into the human liver microsome(rat liver microsome; human recombinant UGT1A1 enzyme) to test the hepatotoxicity using the bilirubin as UGT1A1 enzyme substrate, investigating the inhibition of the UGT1A1 enzyme(in vitro). Apparent inhibition constant Ki and enzyme kinetic parameters were used to evaluate the hepatotoxicity. RESULTS Polygonum multiflorum extract has a strong inhibiton to the UGT1A1 enzyme in all the three systems in vitro. All the type of inhibition is the competitive inhibition. While Polygonum multiflorum extract has a strong inhibiton to the UGT1A1 enzyme in vivo, but the type of inhibition is the uncompetitive inhibition.CONCLUSION The method we had established in our study provides a new idea and a new method to evaluate the hepatotoxicity and the safety of Chinese herbs.
关键词
何首乌提取物 /
肝毒性 /
代谢酶 /
肝微粒体 /
表观抑制常数 /
酶动力学参数 /
胆红素
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Key words
Polygonum multiflorum extract /
hepatotoxicity /
metabolic enzymes /
human liver microsome /
apparent inhibition constant /
enzyme kinetic parameter /
bilirubin
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中图分类号：
R969
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脚注
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基金
国家自然科学基金资助项目(81503347)
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