大黄素-大黄素甲醚型二蒽酮化合物安全性研究

Safety of Emodin-emodin Physcion Dianthrones

目的:评价大黄素-大黄素甲醚型二蒽酮化合物的潜在肝毒性风险。方法:采用计算机分子对接构建尿苷二磷酸-葡萄糖醛酸转移酶1A1 (UGT1A1)与反式-大黄素-大黄素甲醚二蒽酮(trans-EPD)、顺式-大黄素-大黄素甲醚二蒽酮(cis-EPD)的结合模型,考察化合物与酶蛋白的结合位点及结合强弱;体外采用大鼠肝微粒体酶抑制实验评价化合物对UGT1A1的抑制作用;通过细胞增殖与活性检测试剂盒-8 (CCK-8)评价化合物对肝细胞活力的影响;采用荧光定量聚合酶链式反应(qRT-PCR)实验检测肝细胞中UGT1A1 mRNA水平。结果:计算机分子对接实验结果显示,trans-EPD及cis-EPD均与UGT1A1结合于site F区;体外酶抑制实验及细胞毒性实验证明,transEPD和cis-EPD均可显著抑制UGT1A1活性,下调其基因表达,产生肝细胞毒性作用。结论:具有大黄素(10→10′)大黄素甲醚或大黄素(10→10′)大黄素甲醚母核结构的二蒽酮化合物可能是一类具有潜在肝毒性的化合物,其作用靶点为胆红素代谢酶UGT1A1。研究结果为此类成分的临床安全用药提供参考。

Objective: To evaluate the potential hepatotoxicity risk of emodin-emodin physcion dianthrones.Methods: Discovery Studio2.5 was employed for docking of UDP-glucuronyltransferase 1A1(UGT1A1) protein with transemodin-emodin physcion dianthrone(trans-EPD) and cis-emodin-emodin physcion dianthrone(cis-EPD), and the binding sites and intensity were investigated. Rat liver microsomal enzyme inhibition test was used to evaluate the inhibitory effect of the test substances on UGT1A1 in vitro. Cell Counting Kit-8(CCK-8) assay was adopted to examine the influence of them on activity of hepatocytes, and quantitative reverse transcription-polymerase chain reaction(qRT-PCR) to detect the mRNA level of UGT1A1 in hepatocytes.Results: Molecular docking revealed that trans-EPD and cis-EPD bound to UGT1A1 at site F. The in vitro enzyme inhibition experiment and cytotoxicity experiment proved that trans-EPD and cisEPD could significantly suppress the activity of UGT1A1, down-regulate its mRNA level, and induce hepatotoxicity.Conclusion: Dianthrone compounds with emodin(10→10′)-emodin physcion or emodin(10→10′)-emodin physcion nucleus structure have potential hepatotoxicity, and their target is UGT1A1. The findings in this study can serve as a reference for the clinical medication and safety of emodin-emodin physcion dianthrones.