基于UGT1A1抑制作用考察大黄素肝毒性作用

目的：以胆红素代谢过程中UDP-葡萄糖醛酸转移酶1A1（UGT1A1）介导的胆红素葡萄糖醛酸结合环节为切入点，评价大黄素潜在肝毒性风险。方法：以胆红素为UGT1A1底物，以表观抑制常数K_i为评价指标，采用体外肝微粒体孵育法，启动Ⅱ相代谢反应，考察大黄素原型成分的抑制作用，启动Ⅰ、Ⅱ相代谢反应，考察代谢产物及原型成分的综合抑制作用。结果：仅启动Ⅱ相反应时，大黄素以原型形式直接作用于UGT1A1，表现为中强抑制，抑制类型为竞争型抑制；同时启动Ⅰ、Ⅱ两相反应时，大黄素对UGT1A1的抑制作用转为弱抑制，提示大黄素存在Ⅰ相代谢过程，并且其Ⅰ相代谢产物对UGT1A1抑制作用较弱。结论：本实验初步证明大黄素原型对UGT1A1存在抑制作用，经由Ⅰ相代谢后抑制作用降低，肝毒性风险降低。

Objective:To evaluate the potential hepatotoxicity of emodin on the basis of the bilirubin glucuronic acid binding mediated by UDP-glucuronosyltransferase 1A1(UGT1A1) during the process of bilirubin metabolism. Methods:The phase Ⅱ metabolic reaction was initiated by adopting bilirubin as the substrate of UGT1A1 and the in vitro microsome incubation methods,while the apparent inhibition constant K_i was used as the evaluation index. The phase I and Ⅱ metabolic reactions were simultaneously initiated to investigate the combined inhibition effects from metabolites and prototype components. Results:Emodin directly acted on UGT1A1 in the form of a prototype and demonstrated a moderate competitive inhibition,when only the phaseⅡ reaction was initiated;whereas the inhibitory effect of emodin was weaken when both phase I and Ⅱ reactions were initiated,suggesting the participation of emodin in phase I metabolic process,and the inhibitory effect of its phase I metabolites on UGT1A1 was weaker. Conclusion:This experiment preliminarily proves that the emodin prototype could inhibit UGT1A1,which is reduced after phase I metabolism,and the hepatotoxicity risk is decreased correspondingly.

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