苦豆草治疗大肠湿热证大鼠血清代谢组学研究

Sophora alopecuroides ameliorates large intestine dampness-heat syndrome in rats by serum metabolomic approach based on UHPLC-Q/TOF-MS/MS

中药苦豆草具有清肠燥湿的功效,临床用于治疗菌痢、肠炎等疾病,但对于其作用机制尚不完全清楚。本研究以探究苦豆草治疗大肠湿热证作用机制为目的,采用高温高湿环境、高糖高脂饮食结合腹腔注射产肠毒性大肠杆菌的方法建立大肠湿热证模型。通过检测大鼠血常规和促炎细胞因子,观察其肠道组织病理学变化,并结合基于超高效液相色谱与串联四级杆飞行时间二级质谱(UHPLC-Q/TOF-MS/MS)联用技术的血清代谢组学方法进行研究。结果显示,苦豆草能够缓解大肠湿热证症状,改善肠道组织病变及调节体内促炎细胞因子IL-1β、IL-2、IL-6和TNF-α的含量异常。经血清代谢轮廓分析发现牛磺酸、色氨酸、溶血磷脂酰乙醇胺、溶血磷脂酰胆碱、溶血磷脂酸、甘油二酯、鹅去氧胆酸硫酸氢盐、2-十二碳烯二酸和7-酮基脱氧胆酸等9种差异代谢物可能是苦豆草治疗大肠湿热证的潜在代谢标志物,其涉及牛磺酸和亚牛磺酸代谢、甘油磷脂代谢、甘油酯代谢、色氨酸代谢和初级胆酸合成等5条代谢路径。本研究运用血清代谢组学方法揭示了苦豆草干预大肠湿热证的作用机制,为苦豆草临床应用提供了理论依据。

The Chinese herbal Sophora alopecuroides is widely used to clean intestine and eliminate dampness, and it has good therapeutic effects on treating bacillary dysentery and inflammatory bowel disease, etc. in clinics. However, the mechanism of treatment is not yet well understood. The present study was aimed to explore the mechanism of Sophora alopecuroides treatment of large intestine dampness-heat syndrome（LIDHS）. The LIDHS model was performed by the comprehensive factors, including high temperature and humidity environment, high-sugar and high-fat diet, and intraperitoneal injection of Escherichia coli. The blood routine, serum proinflammatory cytokine levels and histopathological changes of intestine were detected and observed. Meanwhile, the serum metabolomic approach was conducted using the method of ultra performance liquid chromatography coupled to quadrupole time-of-flight mass/mass spectrometry（UHPLC-Q/TOF-MS/MS）. The results showed that Sophora alopecuroides has good therapeutic effects on the LIDHS rat models. After treatment with Sophora alopecuroides, the abnormality of blood routine indexes as well as proinflammatory cytokines, including IL-1β, IL-2, IL-6 and TNF-α in vivo, tended to be normal, and the histopathological changes of intestine were improved. Through metabolic profiling and protocol analysis, 9 potential metabolic markers may be closely related with the treatment mechanism of Sophora alopecuroides on this disease, including taurine, L-tryptophan, Lyso PE, Lyso PC, LPA, DG, chenodeoxycholic acid disulfate, traumatic acid and 7-ketodeoxycholic acid, which were involved in taurine and hypotaurine metabolism, glycerophospholipid metabolism, glycerolipid metabolism, tryptophan metabolism and primary bile acid biosynthesis etc. The serum metabolomic approach can be applied to clarify the therapeutic mechanism of Sophora alopecuroides on LIDHS, and provide the theoretical basis for Sophora alopecuroides in clinical practice.