Table_4_Metabolomic Insights Into the Synergistic Effect of Biapenem in Combination With Xuebijing Injection Against Sepsis.docx

The drug combination of biapenem (BIPM) and xuebijing injection (XBJ) is commonly applied for the treatment of sepsis in China. However, the potential synergistic mechanism is still enigmatic. There have been no studies focused on the plasma metabolome alterations in sepsis after the intervention of this combination. In this work, an untargeted metabolomics approach was performed by liquid chromatography-mass spectrometry coupled with multivariate statistical analysis to provide new insights into the synergistic effect of BIPM in combination with XBJ. We characterized the metabolic phenotype of sepsis and described metabolic footprint changes in septic rats responding to XBJ and BIPM individually and in combination, in addition to histopathological and survival evaluation. A total of 91 potential biomarkers of sepsis were identified and 32 disturbed metabolic pathways were constructed. Among these biomarkers, 36 metabolites were reversely regulated by XBJ, mainly including glycerophospholipids, sphingolipids, free fatty acids (FFAs), bile acids and acylcarnitines; 42 metabolites were regulated by BIPM, mainly including amino acids, glycerophospholipids, and acylcarnitines; 72 metabolites were regulated after XBJ-BIPM combination treatment, including most of the 91 potential biomarkers. The results showed that the interaction between XBJ and BIPM indeed exhibited a synergistic effect by affecting some key endogenous metabolites, 15 metabolites of which could not be regulated when XBJ or BIPM was used alone. Compared with Model group, 13, 22, and 27 metabolic pathways were regulated by XBJ, BIPM, and XBJ-BIPM combination, respectively. It suggested that many more endogenous metabolites and metabolic pathways were significantly regulated after combination treatment compared with XBJ or BIPM monotherapy. Metabolisms of lipids, amino acids, acylcarnitines, and bile acids were common pathways involved in the synergistic action of XBJ and BIPM. This study was the first to employ metabolomics to elucidate the synergistic effect and decipher the underlying mechanisms of BIPM in combination with XBJ against sepsis. The results provide some support for clinical application of antibiotics in combination with traditional Chinese medicines and have important implications for the treatment of sepsis in clinic.

比阿培南（biapenem, BIPM）与血必净注射液（xuebijing injection, XBJ）的联合用药方案在我国常用于脓毒症（sepsis）的临床治疗，但其潜在的协同作用机制仍未阐明。目前尚无针对该联合用药干预后脓毒症患者血浆代谢组变化的相关研究。本研究采用液相色谱-质谱联用（liquid chromatography-mass spectrometry, LC-MS）结合多变量统计分析（multivariate statistical analysis）的非靶向代谢组学（untargeted metabolomics）方法，为解析比阿培南与血必净注射液的协同药效机制提供新的研究视角。本研究不仅对脓毒症的代谢表型（metabolic phenotype）进行了表征，还分别考察了血必净注射液、比阿培南单一给药及二者联合给药对脓毒症大鼠的代谢足迹变化，并同步开展了组织病理学评价与生存分析。研究共鉴定出91个脓毒症潜在生物标志物（potential biomarkers），并构建了32条紊乱代谢通路（disturbed metabolic pathways）。在上述生物标志物中，血必净注射液可反向调控36种代谢物，主要涵盖甘油磷脂（glycerophospholipids）、鞘磷脂（sphingolipids）、游离脂肪酸（free fatty acids, FFAs）、胆汁酸及酰基肉碱（acylcarnitines）类物质；比阿培南可调控42种代谢物，主要包括氨基酸、甘油磷脂及酰基肉碱类物质；而二者联合给药后可调控72种代谢物，覆盖上述91种潜在生物标志物中的绝大多数。结果表明，血必净注射液与比阿培南的联用确实可通过影响关键内源性代谢物发挥协同作用，其中15种代谢物仅在联合给药时被调控，单一使用血必净注射液或比阿培南都无法对其产生调节效果。与模型组（Model group）相比，血必净注射液、比阿培南单一给药及联合给药分别调控了13、22及27条代谢通路。这提示相较于单一用药（monotherapy），联合给药可显著影响更多的内源性代谢物与代谢通路。脂类、氨基酸、酰基肉碱及胆汁酸代谢是二者协同作用共同涉及的核心代谢通路。本研究首次采用代谢组学技术阐明了比阿培南与血必净注射液联合治疗脓毒症的协同效应并解析了其潜在作用机制，研究结果为抗生素联合中药（traditional Chinese medicines）的临床应用提供了理论支撑，同时对脓毒症的临床治疗具有重要指导意义。