Effect of Pudilan Xiaoyan Oral Liquid on Energy Metabolism in Mice with Acute Pneumonia Induced by LPS Based on Serum Targeted Metabolomics
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摘要: 目的 研究蒲地蓝消炎口服液对脂多糖(LPS)诱导的急性肺炎小鼠能量代谢的影响。方法 取ICR小鼠随机分为对照组, 模型组, 蒲地蓝消炎口服液低、中、高剂量组和阳性药组, 灌胃给药, 连续给药5 d。GC-MS靶向检测小鼠血清中糖酵解和三羧酸(TCA)循环的12个代谢物(丙酮酸、乳酸、琥珀酸、富马酸、苹果酸、α-酮戊二酸、磷酸烯醇式丙酮酸、顺式乌头酸、3-磷酸甘油酸、柠檬酸、异柠檬酸、葡萄糖), 并进行多变量数据分析。结果 和对照组比较, 模型组小鼠血清中鉴定出乳酸、琥珀酸、苹果酸、α-酮戊二酸、异柠檬酸和葡萄糖等多个显著性差异代谢物, 并在蒲地蓝消炎口服液干预下得以有效改善。结论 研究精准发现与炎症反应相关的6个显著差异代谢物, 为从能量代谢层面上深入阐明蒲地蓝消炎口服液抗呼吸系统炎症的作用机制提供了参考。Abstract: OBJECTIVE To study the effect of Pudilan Xiaoyan Oral Liquid on energy metabolism in mice with lipopolysaccharide(LPS)-induced acute pneumonia.METHODS ICR mice were randomly divided into control group, model group, Pudilan group and positive drug group, and the drugs were given orally for 5 days. GC-MS was used to detect 12 metabolites(pyruvic acid, lactic acid, succinic acid, fumaric acid, malic acid, α-ketoglutarate, phosphoenolpyruvic acid, cis-aconitic acid, 3-phosphoglyceric acid, citric acid, isocitric acid, glucose) in glycolysis and tricarboxylic acid(TCA) cycle in mouse serum and then multivariate data analysis was carried out to search the significant metabolites.RESULTS Among the targeted metabolites, lactic acid, succinic acid, malic acid, α-ketoglutarate, isocitrate and glucose were found as the significant metabolites. Compared to the model group, these metabolites were effectively restored by Pudilan Xiaoyan Oral Liquid.CONCLUSION This study accurately finds six differential metabolites related to inflammation, providing a reference for in-depth explanation of the mechanism of anti-respiratory inflammation of Pudilan Xiaoyan Oral Liquid at the level of energy metabolism.
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Key words:
- Pudilan Xiaoyan Oral Liquid /
- acute pneumonia /
- targeted metabolomics /
- GC-MS /
- energy metabolism
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表 1 靶向代谢物及内标的SRM分析参数
化合物中文名 化合物英文名 保留时间/min 母离子m/z 子离子m/z 碰撞能量/eV 丙酮酸 Pyruvate 4.01 174.1 74.1 14 乳酸 Lactate 4.09 191.2 147.1 8 琥珀酸 Succinate 5.83 147.1 73.1 14 富马酸 Fumarate 6.04 245.1 73.1 18 苹果酸 Malate 6.99 233.1 73.1 10 α-酮戊二酸 α-Ketoglutarate 7.52 198.1 73.1 12 磷酸烯醇式丙酮酸 Phosphoenolpyruvate 7.70 369.1 147.1 16 顺式乌头酸 cis-Aconitate 8.50 229.1 147.1 10 3-磷酸甘油酸 3-Phosphoglycerate 8.88 227.1 211.1 8 柠檬酸 Citrate 8.94 273.1 73.1 18 异柠檬酸 Isocitrate 8.98 245.1 73.1 16 葡萄糖 Glucose 9.42 319.2 73.1 18 1, 2-13C2-肉豆蔻酸(内标) 1, 2-13C2-myristic acid (IS) 9.02 119.1 75.0 10 
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表 2 各组小鼠血清中IL-10、TNF-α和NF-κB水平测定(x±s，pg·mL-1，n=6)
组别 IL-10 TNF-α NF-κB 对照组 93.04±4.524 111.80±3.378 923.1±29.07 模型组 80.19±2.562# 151.10±3.441## 1106.0±39.97## PDL高剂量组 108.70±4.828** 113.20±5.129** 886.5±30.47** PDL中剂量组 95.39±3.047** 129.30±3.385** 973.7±23.32* PDL低剂量组 97.96±2.167** 130.60±2.586** 961.4±33.49* 拜阿司匹林组 93.44±2.301** 141.50±3.784 987.2±26.73* 注：与对照组比较，*P < 0.05，**P < 0.01；与模型组比较，#P < 0.05，##P < 0.01。
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表 3 各组小鼠血清中靶向代谢物的比较
序号 代谢物 模型组vs对照组 给药组vs模型组 VIP值 变化趋势 变化趋势 1 丙酮酸(Pyruvate) 0.924 8 ↑* ↓ 2 乳酸(Lactate) 1.149 8 ↑** ↓* 3 琥珀酸(Succinate) 1.102 7 ↑* ↓** 4 富马酸(Fumarate) 0.962 9 ↑* ↓ 5 苹果酸(Malate) 1.033 2 ↑** ↓** 6 α-酮戊二酸(α-Ketoglutarate) 1.000 9 ↑* ↓** 7 磷酸烯醇式丙酮酸(PEP) 0.769 8 ↑ ↓ 8 顺式乌头酸(cis-Aconitate) 0.816 8 ↑* ↓ 9 3-磷酸甘油酸(3-Phosphoglycerate) 0.914 9 ↑* ↓ 10 柠檬酸(Citrate) 0.911 6 ↑* ↓ 11 异柠檬酸(Isocitrate) 1.011 0 ↑* ↓* 12 葡萄糖(Glucose) 1.288 5 ↑** ↓* 注：*P < 0.05，**P < 0.01。
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