目的 分析鲜生地-淡豆豉同打炮制和配伍水煎液成分差异，比较两组药物改善血热模型大鼠的功效差异，并基于血浆和尿液代谢组学探究其清热凉血及同打增效的潜在作用机制。方法 基于超高效液相色谱-四极杆-静电场轨道阱高分辨质谱（ultra high performance liquid chromatography Q-exactive HF-X，UHPLC-Q-Exactive HF-X）技术分析水煎液成分差异。采用干酵母结合无水乙醇复制血热模型，监测各组大鼠体质量及肛温变化，腹主动脉取血测定各切变率下全血黏度及凝血四项，HE染色观察肺、胃组织的病理学变化。基于超高效液相色谱-四极杆-飞行时间串联质谱（ultra high performance liquid chromatography quadrupole-time of flight tandem mass spectrometry，UHPLC-Q-TOF-MS/MS）技术结合多元统计方法筛选大鼠血浆和尿液中与血热证相关的潜在生物标记物，并分析同打组与配伍组改善血热证的代谢调控机制及差异。结果 同打组水煎液共检测出66种成分，配伍组水煎液共检测出57种成分。给药干预后，血热大鼠的体质量增长率显著上升、肛温明显下降、各切变率下的全血黏度降低、凝血酶原时间（prothrombin time，PT）显著延长而活化部分凝血活酶时间（activated partial thromboplastin time，APTT）显著缩短，肺、胃组织的炎症细胞浸润及出血等病理变化显著改善。其中，与配伍组相比，同打组对肛温、中高切变率下的全血黏度及肺胃组织的改善作用明显更佳。代谢组学结果显示，在血热模型大鼠血浆及尿液中共鉴定出55种潜在生物标志物，涉及8条代谢通路。药物干预后，配伍组中有20种发生显著回调，涉及α-亚麻酸代谢、三羧酸循环和类固醇激素生物合成代谢通路；而同打组中有26种发生显著回调，涉及5条代谢通路。结论 淡豆豉鲜生地同打可能通过调节鞘脂代谢及色氨酸代谢通路增强其清热凉血作用，为鲜生地淡豆豉同打在中医临床上的应用提供了科学依据和合理支撑。

Abstract: Objective To analyze the differences of components between co-processed fresh Rehmanniae Radix-Sojae Semen Praeparatum (co-processing group) and the compatibility group, compare the efficacy differences and compatibility in improving blood-heat model rats, and explore the potential mechanism of both clearing heat and cooling blood effects and the synergistic effect of co-processing based on plasma and urine metabolomics. Methods Ultra high performance liquid chromatography Q-exactive HF-X (UHPLC-Q-Exactive HF-X) technology was employed to analyze compositional differences in the water decoctions. Rats were subcutaneously injected with dried yeast and given anhydrous ethanol to create a blood-heat model. Body mass and anal temperature of rats in each group were monitored. Blood was collected from the abdominal aorta to determine the whole blood viscosity and four coagulation parameters. Hematoxylin and eosin (HE) staining was used to observe the pathological changes in lung and stomach tissues. Ultra high performance liquid chromatography quadrupole-time of flight tandem mass spectrometry (UHPLC-Q-TOF-MS/MS) technology combined with multivariate statistical methods were employed to screen potential biomarkers related to blood-heat syndrome in plasma and urine samples. The metabolic regulation mechanisms and their differences of the processing group and the compatibility group n improving blood heat syndrome were analyzed. Results A total of 66 chemical components were detected in the co-processed group decoction, while 57 components were identified in the compatibility group decoction. After the intervention, the body mass growth rate of the diseased rats increased significantly, the anal temperature decreased significantly, the whole blood viscosity decreased at all shear rates, the PT time was prolonged while the APTT time was shortened, and the pathological changes such as inflammatory cell infiltration and hemorrhage in the lungs and stomach tissues were improved significantly. In particular, compared with the compatibility group, the processing group showed significantly better improvement in anal temperature, whole blood viscosity at medium and high shear rates, and pathological changes in lung and stomach tissues. Metabolomics results showed that 55 potential biomarkers were identified in plasma and urine of blood-heat model rats, involving eight metabolic pathways. After the intervention, 20 biomarkers were significantly regulated in the compatibility group, involving α-linolenic acid metabolism, steroid hormone biosynthesis, and citrate cycle metabolism pathways. Whereas 26 biomarkers were regulated considerably in the processing group, involving five metabolism pathways. Conclusion Fresh Rehmannia Radix processed with Semem Sojae Praeparatum may enhance its clearing heat and cooling blood effect by regulating sphingolipid metabolism and tryptophan metabolism pathways, which provides a scientific basis and reasonable support for the clinical application of fresh Rehmanniae Radix processed with Sojae Semen Praeparatum in traditional Chinese medicine.

R284；R286

国家自然科学基金青年项目（82204621）；南京中医药大学国自然青年基金经费配套项目（XPT82204621）；南京中医药大学中药炮制技术传承基地建设项目