大黄素-8-O-β-D-葡萄糖苷及其代谢产物肝毒性研究  

Study on hepatotoxicity of emodin-8-O-β-D-glucoside and its metabolites

作　　者：汪祺[1] 王亚丹[1] 杨建波[1] 刘越[1] 文海若 马双成[1] WANG Qi;WANG Ya-dan;YANG Jian-bo;LIU Yue;WEN Hai-ruo;MA Shuang-cheng(National Institutes for Food and Drug Control,Beijing 100050,China)

机构地区：[1]中国食品药品检定研究院,北京100050

出　　处：《中国新药杂志》2020年第16期1856-1862,共7页Chinese Journal of New Drugs

基　　金：国家自然基金资助项目(81503347,81773874);国家“重大新药创制”科技重大专项资助项目(2018ZX09735-006):中药组分资源库及产业公共技术服务平台建设。

摘　　要：目的:采用HepaRG细胞系及体外肝微粒体体系初步评价及推测大黄素-8-O-β-D-葡萄糖苷(EG)的潜在肝毒性风险。方法:通过三磷酸腺苷(ATP)生物发光法测定EG对HepaRG细胞的抑制率评价EG的整体细胞毒作用;采用体外肝微粒体孵育法,启动Ⅱ相代谢反应,考察EG原型对尿苷二磷酸葡萄糖醛酸转移酶1A1(UGT1A1)的抑制作用;启动Ⅰ和Ⅱ相代谢反应,考察EG代谢产物的潜在肝毒性。综合细胞毒性实验及UGT1A1酶抑制结果初步评价EG的肝毒性作用。结果:HepaRG细胞实验结果显示,EG具有肝细胞毒性作用;UGT1A1酶抑制实验表明,EG原型及其代谢产物均可显著抑制UGT1A1酶活性,且抑制类型均为竞争型抑制,酶活性显著降低可导致其底物胆红素体内蓄积,继而存在引发肝毒性的可能。结论:大黄素-8-O-β-D-葡萄糖苷及其经Ⅱ相代谢酶UDP-葡萄糖醛酸转移酶1A1产生的Ⅱ相代谢物,可能是产生肝脏毒性的原因,二者可通过抑制UGT1A1酶引发胆红素体内堆积产生肝毒性。Objective: To evaluate the potential hepatotoxicity risk of emodin-8-O-β-D-glucoside (EG) by HepaRG cell and liver microsome system in vitro. Methods: The toxic effect of EG on HepaRG cells was determined by Adenosine triphosphate (ATP) bioluminescence. In vitro liver microsomes incubation was used to initiate the phase Ⅱ metabolic reaction to investigate the inhibitory effect of EG on uridine diphosphate glucuronyltransferase 1A1 (UGT1A1) enzyme. The phase I and phase Ⅱ metabolic reactions were initiated to investigate the hepatotoxicity risk of EG metabolites. Results: HepaRG cell assay showed that EG had hepatotoxicity;UGT1A1 inhibition assay showed that EG prototype and its metabolites could significantly inhibit UGT1A1 enzyme activity,and the inhibition type was competitive inhibition. As the enzyme activity was significantly reduced the substrate bilirubin accumulated in the body,which in turn may cause hepatotoxicity. Conclusion: The in vitro toxicity test and enzyme inhibition experiment results showed that EG has hepatotoxicity risk. The potentially toxic substances are its prototype and metabolites emodin monoglucoside and emodin glucuronide glucoside conjugate. The inhibition of UGT1A1 enzyme,which causes accumulation of bilirubin thus leading to hepatotoxicity in vivo,may be the toxicity mechanism of EG.

关 键 词：大黄素-8-O-β-D-葡萄糖苷 UDP-葡萄糖醛酸转移酶1A1 肝代谢 代谢产物 

分 类 号：R992[医药卫生—毒理学]