Effect of Pudilan Xiaoyan Oral Liquid on Energy Metabolism in Mice with Acute Pneumonia Induced by LPS Based on Serum Targeted Metabolomics
-
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.
-
Key words:
- Pudilan Xiaoyan Oral Liquid /
- acute pneumonia /
- targeted metabolomics /
- GC-MS /
- energy metabolism
-
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 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。 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。 -
[1] 国家药典委员会. 中华人民共和国药典: 一部[S]. 北京: 中国医药科技出版社, 2015. [2] 童小慧, 郑悦, 黄玮玲, 等. 蒲地蓝消炎制剂药理及临床研究进展[J]. 云南中医学院学报, 2018, 41(3): 98-102. https://www.cnki.com.cn/Article/CJFDTOTAL-YNZY201803022.htm [3] 吴海燕. 蒲地蓝消炎口服液对急性上呼吸道感染患儿炎性因子及免疫功能的影响[J]. 中国妇幼保健, 2020, 35(11): 2037-2040. https://www.cnki.com.cn/Article/CJFDTOTAL-ZFYB202011027.htm [4] 白玉, 李宇翔, 时宇静, 等. 蒲地蓝消炎口服液治疗小儿上呼吸道感染临床效果及安全性的Meta分析[J]. 中国中药杂志, 2020, 45(9): 2203-2209. https://www.cnki.com.cn/Article/CJFDTOTAL-ZGZY202009033.htm [5] 杨依霏, 张广平, 高云航, 等. 蒲地蓝消炎口服液对脂多糖致大鼠急性肺损伤的影响[J]. 中国实验方剂学杂志, 2019, 25(13): 55-59. https://www.cnki.com.cn/Article/CJFDTOTAL-ZSFX201913009.htm [6] 刘江华, 郑晓文, 温保强. LPS诱导的急性肺损伤中早期标志物表达研究[J]. 蛇志, 2018, 30(3): 391-395, 398. doi: 10.3969/j.issn.1001-5639.2018.03.001 [7] 张子婧, 蒲诗云, 何金汗. 炎性小体在能量代谢中的作用[J]. 生理科学进展, 2018, 49(6): 451-455. doi: 10.3969/j.issn.0559-7765.2018.06.011 [8] ZHAI Y, XU J, FENG L, et al. Broad range metabolomics coupled with network analysis for explaining possible mechanisms of Er-Zhi-Wan in treating liver-kidney Yin deficiency syndrome of Traditional Chinese medicine[J]. J Ethnopharmacol, 2019, 234: 57-66. doi: 10.1016/j.jep.2019.01.019 [9] ZHU JJ, DJUKOVIC D, DENG LL, et al. Targeted serum metabolite profiling and sequential metabolite ratio analysis for colorectal cancer progression monitoring[J]. Anal Bioanal Chem, 2015, 407(26): 7857-7863. doi: 10.1007/s00216-015-8984-8 [10] XU J, ZHAI Y, FENG L, et al. An optimized analytical method for cellular targeted quantification of primary metabolites in tricarboxylic acid cycle and glycolysis using gas chromatography-tandem mass spectrometry and its application in three kinds of hepatic cell lines[J]. J Pharm Biomed Anal, 2019, 171: 171-179. doi: 10.1016/j.jpba.2019.04.022 [11] 张琪, 董旭阳, 许秀丽, 等. 基于靶向代谢组学法鉴别真伪海鸭蛋[J]. 中国食品卫生杂志, 2020, 32(3): 250-256. https://www.cnki.com.cn/Article/CJFDTOTAL-ZSPZ202003010.htm [12] 刘芳. 蒲地蓝口服液辅助匹多莫德对反复呼吸道感染患儿(附119例临床资料)临床疗效评价[J]. 抗感染药学, 2020, 17(4): 593-595. https://www.cnki.com.cn/Article/CJFDTOTAL-KGYY202004049.htm [13] NUNES-NESI A, ARAÚJO WL, OBATA T, et al. Regulation of the mitochondrial tricarboxylic acid cycle[J]. Curr Opin Plant Biol, 2013, 16(3): 335-343. doi: 10.1016/j.pbi.2013.01.004 [14] BOUMEZBEUR F, BESRET L, VALETTE J, et al. Glycolysis versus TCA cycle in the primate brain as measured by combining 18F-FDG PET and 13C-NMR[J]. J Cereb Blood Flow Metab, 2005, 25(11): 1418-1423. doi: 10.1038/sj.jcbfm.9600145 [15] TAVSAN Z, AYAR KAYALI H. The variations of glycolysis and TCA cycle intermediate levels grown in iron and copper mediums of Trichoderma harzianum[J]. Appl Biochem Biotechnol, 2015, 176(1): 76-85. doi: 10.1007/s12010-015-1535-0 [16] SHIMODA Y, HAN J, KAWADA K, et al. Metabolomics analysis of Cistus monspeliensis leaf extract on energy metabolism activation in human intestinal cells[J]. J Biomed Biotechnol, 2012, 2012: 1-7. http://www.scienceopen.com/document_file/66fe59e7-975c-43ba-b756-ecb4ed27490b/PubMedCentral/66fe59e7-975c-43ba-b756-ecb4ed27490b.pdf [17] 蒋龙元, 周天恩, 张萌, 等. 乳酸清除率和乳酸脱氢酶对急性肺损伤预后的评估[J]. 中华危重症医学杂志(电子版), 2010, 3(2): 90-94. doi: 10.3969/cma.j.issn.1674-6880.2010.02.004 [18] 刘义鑫, 潘灵辉, 林飞, 等. 肺组织低氧代谢中葡萄糖转运蛋白1的表达及其作用[J]. 重庆医学, 2012, 41(9): 872-874, 877. doi: 10.3969/j.issn.1671-8348.2012.09.015 [19] TANNAHILL GM, CURTIS AM, ADAMIK J, et al. Succinate is an inflammatory signal that induces IL-1β through HIF-1α[J]. Nature, 2013, 496(7444): 238-242. doi: 10.1038/nature11986 [20] 华永丽. 当归挥发油干预大鼠LPS炎症模型的相关代谢物及代谢通路分析[D]. 兰州: 甘肃农业大学, 2014. [21] HO WE, XU YJ, XU FG, et al. Metabolomics reveals altered metabolic pathways in experimental asthma[J]. Am J Respir Cell Mol Biol, 2013, 48(2): 204-211. doi: 10.1165/rcmb.2012-0246OC [22] WOLAK JE, ESTHER CR, O'CONNELL TM. Metabolomic analysis of bronchoalveolar lavage fluid from cystic fibrosis patients[J]. Biomarkers, 2009, 14(1): 55-60. doi: 10.1080/13547500802688194 [23] DE CARVALHO LPS, FISCHER SM, MARRERO J, et al. Metabolomics of Mycobacterium tuberculosis reveals compartmentalized co-catabolism of carbon substrates[J]. Chem Biol, 2010, 17(10): 1122-1131. doi: 10.1016/j.chembiol.2010.08.009 [24] MILLS EL, O'NEILL LA. Reprogramming mitochondrial metabolism in macrophages as an anti-inflammatory signal[J]. Eur J Immunol, 2016, 46(1): 13-21. doi: 10.1002/eji.201445427