Additional file 4: of Gut microbiota-mediated generation of saturated fatty acids elicits inflammation in the liver in murine high-fat diet-induced steatohepatitis

Schematic illustration of the substances detected in the metabolomic analysis of the metabolic pathways. (A) Global metabolomic profiling comparing the detectable molecules in the feces among the 3 experimental groups was performed (N = 3 in each group) to determine how different gut bacteria metabolize food. Lipid metabolites in the feces were analyzed using liquid chromatography time-of-flight mass spectrometry (LC-TOFMS), and hydrophilic metabolites were analyzed by capillary electrophoresis time-of-flight mass spectrometry (CE-TOFMS). We identified 225 peaks (158 cations and 67 anions) of hydrophobic metabolites by CE-TOFMS, 115 peaks (65 positives and 50 negatives) of hydrophilic metabolites by LC-TOFMS, and 340 candidate compounds (CE-TOFMS 225 and LC-TOFMS 115). These detected peaks were categorized into glycolysis/glyconeogenesis, pentose-phosphate, tricarboxylic acid (TCA) cycle, urea cycle, purine-pyrimidine, coenzyme, amino acids, acyl-carnitine, and fatty acid pathways and were included in a pathway map. Pathway mapping shows a quantitative comparison of the molecules in the 3 experimental groups. (B) The 38 selected metabolites that were increased specifically in antibiotics treated group compared to the control or STHD-01 groups. (N = 3 in each group) Among these metabolites, 6 metabolites were detected in STHD-01 + Abx group in high concentration, while these were undetectable in the STHD-01 group. The concentration of 32 metabolites were as >3-fold higher in STHD-01 + Abx group than that in the STHD-01 group. (C) The 78 selected metabolites that were increased specifically in the STHD-01 group compared to the STHD-01 + Abx group. (N = 3 in each group) Among these metabolites, 16 metabolites were detected in the STHD-01 group in high concentration, while these were undetectable in the STHD-01 + Abx group. The concentration of 62 metabolites were as >3-fold higher in the STHD-01 group than that in the STHD-01 + Abx group. (ZIP 552 kb)

本示意图展示了代谢通路代谢组学分析中所检测到的物质。(A) 本研究针对3组实验对象的粪便样本开展了可检测分子的全局代谢组学分析（每组n=3），以探究不同肠道菌群对食物的代谢模式。粪便中的脂质代谢物采用液相色谱-飞行时间质谱（LC-TOFMS）进行分析，亲水性代谢物则采用毛细管电泳-飞行时间质谱（CE-TOFMS）进行检测。通过CE-TOFMS共鉴定出疏水性代谢物的225个峰（含158个阳离子峰与67个阴离子峰），通过LC-TOFMS鉴定出亲水性代谢物的115个峰（含正离子模式65个、负离子模式50个），总计获得340个候选化合物（其中CE-TOFMS鉴定225个、LC-TOFMS鉴定115个）。上述检测到的峰被归类为糖酵解/糖异生通路、戊糖磷酸通路、三羧酸（TCA）循环、尿素循环、嘌呤-嘧啶通路、辅酶代谢通路、氨基酸代谢通路、酰基肉碱通路以及脂肪酸代谢通路，并被整合至通路图谱中。该通路图谱实现了3组实验对象中代谢分子的定量比较。(B) 筛选出38种仅在抗生素（Abx）处理组中上调的代谢物，与对照组或STHD-01组相比（每组n=3）。其中，6种代谢物在STHD-01+Abx组中呈高浓度表达，而在STHD-01组中未被检测到；另有32种代谢物在STHD-01+Abx组中的浓度较STHD-01组升高3倍以上。(C) 筛选出78种仅在STHD-01组中上调的代谢物，与STHD-01+Abx组相比（每组n=3）。其中，16种代谢物在STHD-01组中呈高浓度表达，而在STHD-01+Abx组中未被检测到；另有62种代谢物在STHD-01组中的浓度较STHD-01+Abx组升高3倍以上。（压缩包大小：552 KB）