基于化学轮廓方法的UGT酶介导黄连降低大黄多酚类成分生物利用度的药动学机制研究

According to the UGT metabolism and enterohepatic circulation features of polyphenols, the mechanisms of the reduced system exposures of polyphenols of Rhubarb after compatibility of coptis chinensis mediated by UGT in the key link (intestinal, enterohepatic circulation, intestinal flora) were investigated in this study. In vivo pharmacokinetic model, in situ intestinal/liver perfusion model, in vitro UGT metabolism model and fecal flora hatch model were integrated to evaluate the correlation of pharmacokinetics of the total polyphenols and the UGT metabolism features of UGT metabolites profilings. In this study, chemical profiling combination with multivariate data analysis technique (PCA, PLS-DA) were applied to investigate the pharmacokinetics difference of total polyphenols, the metabolic differences of metabolites profile of UGT metabolites. At the same time, the quantitative activity of UGT enzymes in in vitro UGT metabolism model were analyzed to reveal the effect of coptis on the UGT metablism of polyphenols. The results was used to clarify pharmacokinetic interaction mechanisms of polyphenols after compatibility of coptis chinensis mediated by UGT. In addition, chemical profiling combination with multivariate data analysis technique (PCA, PLS-DA) was also applied to investigate the effect of intestinal flora on the in vitro UGT metabolites. Combining the activity of beta glucuronic acid enzyme of intestinal flora before and after compatibility of coptis chinensis, the role of the intestinal flora in the UGT combination reversible and its enterohepatic circulation of polyphenols was also clarified in this study. Thus, the drug interaction including the phase II metabolic interactions and interactions of removing the UGT metabolite in the enterohepatic circulation by intestinal flora was expounded mediated by UGT enzyme. These results can clarify the pharmacokinetic mechanism of reducing bioavailability of polyphenols of Rhubarb after compatibility of Coptis mediated by UGT enzymes. At the same time, it indicated that the chemical profiling combination with multivariate data analysis technique would be applied to overall pharmacokinetics study of traditional Chinese medicine compatibility.

前期研究表明黄连显著降低大黄多酚类成分生物利用度，结合多酚类成分广泛的UGT酶代谢及肠肝循环特征，选择经典药对大黄-黄连为模型药物，采用化学轮廓方法，围绕UGT作用的肠道、肠肝循环及肠道菌群等关键环节，开展UGT介导的黄连降低大黄多酚类成分生物利用度的药动学机制研究。项目整合体内药动学模型—大鼠原位肠肝灌注模型—体外UGT代谢模型—体外菌群温孵等模型，采用反映中药成分组特征的化学轮廓方法，结合多变量分析技术（PCA、PLS-DA），获得以各总多酚成分为指标的药动学差异特征；以代谢物轮廓为指标的UGT代谢差异特征及菌群介导的脱UGT化差异特征。通过药动学与UGT代谢相互作用的相关性分析，结合配伍黄连前后的UGT酶及β-葡萄糖醛酸酶的活性的分析，阐明UGT介导的大黄、黄连体内相互作用的药动学机制。项目有利于多学科的交叉渗透，为符合中药整体观特征的中药复方药动学研究提供方法学借鉴。

课题围绕UGT介导的黄连降低大黄蒽醌类成分药动学机制为中心，比较了大黄、黄连配伍前后水煎煮液（以及大黄不同提取方法）的蒽醌类成分游离态和结合态的化学轮廓特征差异；阐明了整体动物模型和大鼠原位肠肝灌注模型下大黄、黄连配伍前后蒽醌类成分的体内药动学特征；阐明了UGT酶介导下黄连活性成分（盐酸小檗碱）、黄芩活性成分（黄芩素）对大黄活性成分（大黄酚）的UGT代谢影响及相关机制；论述了糖尿病大鼠模型与正常大鼠模型下黄芩、黄连药效成分对大黄药效成分药动学特征的影响及与UGT酶相关的相应机制，开展了菌群紊乱大鼠模型下黄连、黄芩和大黄药效成分配伍前后的药动学差异及相关机制；新增加泻心汤（大黄、黄连、黄芩）对糖尿病大鼠肝胆汁代谢影响、药效作用及相关机制。项目已发表7篇科研论文，其中SCI收录2篇，培养了博士后（访问学者)1名，硕士研究生3名，本科生2名。结果显示，大黄蒽醌类成分大黄酚在配伍黄芩素、盐酸小檗碱、黄芩苷后体内药动学参数显著改变，结合菌群紊乱模型下盐酸小檗碱对大黄酚药动学特征影响较小而黄芩素类成分影响显著的结果，大黄药效成分与黄芩素配伍后呈现UGT代谢性底物浓度竞争抑制特征，揭示UGT酶介导是大黄蒽醌类成分配伍黄连或黄芩体内生物利用度降低的重要机制之一。项目围绕大黄配伍黄连后UGT酶介导作用及机制展开多层次的研究，阐明UGT介导的大黄、黄连（新增黄芩）药效成分体内相互作用的药动学机制，为中药复方药物配伍体内代谢药动学相互作用及机制研究提供方法学借鉴与参考。