基于代谢调控通路的京尼平苷干预阳黄证的作用靶标研究

An Exploration in the Action Targets for the Avail of Geniposide Against Yang Huang Syndrome Based on the Regulation of Metabolic Pathways

目的:基于代谢调控通路技术对京尼平苷干预阳黄证小鼠进行研究,探究阳黄证的发病机制及京尼平苷对其干预作用的潜在效应靶标。方法:采用化学物质、中药及病理性肝损伤因素建立阳黄证动物模型,通过与临床阳黄证患者尿液的生物标记物相关联,成功建立阳黄证动物模型。在此基础上对生化指标、病理观察进行检测,基于代谢组学技术挖掘京尼平苷干预阳黄证的作用靶标。结果:与空白组相比,模型组小鼠谷草转氨酶、谷丙转氨酶、总胆红素、总胆汁酸含量升高,具有显著性差异(P<0.05)。碱性磷酸酶、直接胆红素、丙二醛、谷氨酰转肽酶含量升高,谷胱甘肽过氧化酶、超氧化物歧化酶含量降低;病理结果显示,模型组小鼠肝组织成片状坏死,肝细胞有水肿现象。对模型组小鼠尿液进行基于代谢调控的靶点分析,发现33个尿液生物标记物与阳黄证的发生密切相关。其中,10个生物标记物与临床阳黄证患者生物标记物变化一致,主要涉及酪氨酸代谢、牛磺酸和亚牛磺酸代谢、谷胱甘肽代谢、戊糖和葡萄糖醛酸酯代谢、初级胆汁酸代谢等。进一步通过Ingenuity Pathway Analysis组学数据分析平台成功预测京尼平苷干预阳黄证的5个作用靶点,即干扰素调节因子3、鸟氨酸脱羧酶抗酶1(OAZ1)、鸟氨酸脱羧酶1(ODC1)、解偶联蛋白(UCP1)、Maf1调节剂(Maf1 regulator)。结论:本研究以中医证候理论为指导,在对阳黄证形成本质考察的基础上,以中药与肝损伤化学诱导剂结合的方法建立阳黄证动物模型,与临床黄疸生物标记物相关联,发现10个生物标记物与阳黄证发生发展密切相关,京尼平苷可有效回调生物标记物,通过代谢调控通路技术分析发现京尼平苷干预阳黄证的5个潜在作用靶标。本研究为阳黄证相关动物模型建立及中药有效成分治疗作用机制研究提供方法和依据。

The present study aimed at exploring the targets and the pathogenesis of the avail of geniposide against Yang Huang syndrome（YHS） using metabolic technologies. YHS mouse model was established by three methods, including chemicals, Chinese materia medica and pathologic hepatic injury. On this basis, the biomarkers of YHS, various biochemical indexes and pathology observation were adopted to evaluate the modeling of YHS. Compared with the control group, it was found that the contents of ALT, AST, T-Bili and TBA were successfully increased in the model group with significant statistical differences（P0.05）. The levels of ALP, D-Bili, MDA and γ-GT were up-regulated, while the levels of GSH-Px and T-SOD were down-regulated. Besides, pathological observation indicated regional patchy necrosis and hepatic edema emerged in the model mice. Involving metabolic profiling analysis, a total of 33 urinary biomarkers were identified in the model mice, which all associated with YHS, and among which 10 biomarkers were identified that they shared the same structure with the biomarkers of YHS. These biomarkers were chiefly involved in tyrosine metabolism, taurine and hypotaurine metabolism and glutathione metabolism, pentose and glucuronate metabolism, and primary bile acid metabolism. With the global omics analysis of IPA software, we predicted 5 targets engaged in the avail of geniposide against YHS, including OAZ1, ODC1, UCP1 and Maf1 regulator. In conclusion, this study successfully established the YHS mouse model based on the guidance of traditional Chinese medical theory and the essential characteristics of YHS. 10 biomarkers shared the same structure with those of YHS, which played critical roles in the process of the metabolism of YHS. Strikingly, these biomarkers were recovered to the normal levels after the adminstration of geniposide. The aforesaid descriptions indicated that the establishment of the YHS model showed a high consistency with clinical YHS patients. Geniposide may perform favorable efficacy with the aforementioned biomarkers or pathways.