Brain Metabolomics Reveals the Antipyretic Effects of Combined Use of Ling-Zhu-San and Acetaminophen in Young Rats Using Gas Chromatography-Mass Spectrometry
-
摘要:
目的 探讨中成药羚珠散(Ling-Zhu-San, LZS)和西药对乙酰氨基酚(Acetaminophen, APAP)对幼龄大鼠的解热作用及其代谢影响。 方法 采用气相色谱-质谱联用平台、多变量统计技术和通路分析等方法，通过系统的代谢组学分析，确认治疗前后大鼠脑组织中代谢物水平变化；通过体温试验和酶联免疫吸附实验(ELISA)考察大鼠体温变化及白细胞介素-1β (Interleukin-1β，IL-1β)和前列腺素E2 (Prostaglandin E2, PGE2)的含量。 结果 大鼠脑内与TCA循环、色氨酸代谢、谷氨酸代谢和花生酸代谢相关的16种显著改变的代谢物(P < 0.05, FC>1.5)被认为是潜在的发热生物标志物，而联合用药后这些代谢物的浓度均基本恢复至正常水平。羚珠散和对乙酰氨基酚联用较单药解热效果最佳。 结论 羚珠散和对乙酰氨基酚联合用药对幼龄大鼠的解热作用可能与其改善脑内代谢紊乱相关。 Abstract:OBJECTIVE To investigate the antipyretic and metabolic effects of Ling-Zhu-San (LZS) and Acetaminophen (APAP) on young rats. METHODS The changes of metabolite levels in rat brain were confirmed by GC-MS platform, multivariate statistical technique and pathway analysis. The changes of temperature and the contents of IL-1β and PGE2 in rats were investigated by temperature test and enzyme-linked immunosorbent assay. RESULTS 16 metabolites associated with TCA cycle, tryptophan metabolism, glutamate metabolism and arachidonic acid metabolism (P < 0.05, FC>1.5). These metabolites were considered as potential biomarkers of fever. And the concentration of these metabolites basically recovered to normal levels after combination therapy. LZS combined with APAP had the best antipyretic effect. CONCLUSION These results suggest that the antipyretic effect of LZS combined with APAP on young rats may be related to its improvement of brain metabolic disorders. -
Key words:
- Ling-Zhu-San /
- acetaminophen /
- antipyretic effects /
- metabolomics
-
图 2 The changing trends of rectal temperature and relative cytokine concentrations in rat plasma
Note：A. The rectal temperatures were measured at 0, 30, 60, 90, 120, 180 min after drug administration; B, C. The concentration of IL-1β and PGE2 in rat plasma from NG, MG, LZS, CMT and APAP groups. ##P < 0.01, compared with NG; *P < 0.05, * *P < 0.01, compared with MG.
图 3 Typical base peak intensity chromatogram of brain samples from NG, MG, LZS, APAP and CMT group
图 4 PCA and OPLS-DA score plots of the NG, MG, LZS, APAP and CMT group
Note: A. PCA score plot of five groups; B. OPLS-DA score plot of NG, MG, LZS, CMT and APAP group; C. OPLS-DA score plot of NG, MG and LZS group; D. OPLS-DA score plot of NG, MG and CMT group; E. OPLS-DA score plot of NG, MG and APAP group; F. Validation of OPLS-DA model by permutation test (n=200). R2=0.034 8, Q2=-0.318.
图 5 Box and whisker plots of typically altered metabolites in brain samples as measured by GC-MS
Note：The bottom and the top of the box are the 25th and the 75th percentile, a horizontal line near the middle of the box represents the 50th percentile, or the median. #P < 0.05, ##P < 0.01, compared with NG; *P < 0.05, * *P < 0.01, compared with MG.
图 6 The pathways closely associated with the antipyretic and anti-inflammation effects
Note：Metabolites in black bold fonts were potential biomarkers that were affected by CMT.
表 1 Identified chemical materials of LZS
No. Compound TR/min Chemical structure 1 2-Bornanone 5.33 
2 Isoborneol 5.42 
3 endo-Borneol 5.49 
4 α-Copaene 7.02 
5 Cyclohexane 7.11 
6 Methyleugenol 7.14 
7 Benzene 7.49 
8 Aromandendrene 7.62 
9 cis-β-Asarone 8.18 
10 1H-Cycloprop[e]azulen-7-ol 8.34 
11 Eremophila ketone 8.29 
12 Caryophyllene oxide 8.40 
13 Asarone 8.46; 8.82 
14 Junenol 8.62 
15 (1R, 7S, E)-7-Isopropyl-4, 10- dimethylenecyclodec-5-enol 8.98 
16 1-(2, 4, 5-Trimethoxyphenyl) propan-2-one 9.22 
17 Platambin 9.95 
18 2-[4-Methyl-6-(2, 6, 6- trimethylcyclohex-1-enyl) hexa-1, 3, 5-trienyl]cyclohex- 1-en-1-carboxaldehyde 10.15 
19 Cholest-5-en-3-ol 15.30 
20 Lupeol 16.48 
21 Olean-12-en-3-ol 16.70 
22 α-Amyrin 17.02 
23 Ursolic Aldehyde 18.44 
下载: 导出CSV
表 2 Potential pyretic biomarkers
Query HMDB KEGG Fold Change P (NG vs. MG) 1-2-4-Benzenetriol HMDB02708 C05905 2.591 5 < 0.000 1 1-Monostearin HMDB31075 NA 1.204 2 0.001 7 8-245-α-Cholestadien-3-β-ol NA NA 1.226 0 0.020 0 Arachidic acid HMDB02212 C06425 1.294 0 0.001 7 Asparagine HMDB00168 C00152 0.777 5 0.029 5 Aspartate HMDB00191 C00049 1.503 7 0.004 7 Behenic acid HMDB00944 C08281 1.341 9 0.004 6 Cystine HMDB00192 C00491 0.612 5 0.007 3 Histidine HMDB00177 C00135 0.554 2 < 0.000 1 Monoolein NA NA 2.198 2 0.000 6 Oxoproline HMDB00267 C01879 0.798 4 0.038 1 Pyrophosphate HMDB00250 C00013 0.671 6 0.005 0 Ribose HMDB00283 C00121 0.682 4 0.000 8 Tryptophan HMDB00929 C00078 0.601 9 < 0.000 1 Uridine HMDB00296 C00299 1.453 0 0.003 5 Xanthine HMDB00292 C00385 0.769 0 < 0.000 1
下载: 导出CSV
-
[1] KHALIL SN, HAHN BJ, CHUMPITAZI CE, et al. A multicenter, randomized, open-label, active-comparator trial to determine the efficacy, safety, and pharmacokinetics of intravenous ibuprofen for treatment of fever in hospitalized pediatric patients[J]. BMC Pediatr, 2017, 17: 42. doi: 10.1186/s12887-017-0795-y [2] ALSANAD SM, HOWARD RL, WILLIAMSON EM. An assessment of the impact of herb-drug combinations used by cancer patients[J]. BMC Complement Altern Med, 2016, 16: 393. doi: 10.1186/s12906-016-1372-x [3] HE YT, LU AP, ZHA YL, et al. Differential effect on symptoms treated with traditional Chinese medicine and western combination therapy in RA patients[J]. Complementary Ther Med, 2008, 16(4): 206-211. doi: 10.1016/j.ctim.2007.08.005 [4] WANG GQ, XIE YM, WANG LX, et al. Analysis of drug combination characteristics of Qingkailing injection for treating abnormal inflammatory factors in real world[J]. China J Chin Mater Med, 2017, 42(10): 1984-1990. [5] QIN LL, ZHANG ZX, GUO MX, et al. Plasma metabolomics combined with lipidomics profiling reveals the potential antipyretic mechanisms of Qingkailing injection in a rat model[J]. Chem Biol Interact, 2016, 254: 24-33. doi: 10.1016/j.cbi.2016.05.022 [6] GAO XY, GUO MX, LI Q, et al. Plasma metabolomic profiling to reveal antipyretic mechanism of Shuang-Huang-Lian injection on yeast-induced pyrexia rats[J]. PLoS ONE, 2014, 9(6): e100017. doi: 10.1371/journal.pone.0100017 [7] LIU R, HUANG Q, SHAN JJ, et al. Metabolomics of the antipyretic effects of bubali Cornu (water buffalo horn) in rats[J]. PLoS ONE, 2016, 11(7): e0158478. doi: 10.1371/journal.pone.0158478 [8] PANDEY R, CAFLISCH L, LODI A, et al. Metabolomic signature of brain cancer[J]. Mol Carcinog, 2017, 56(11): 2355-2371. [9] ZHANG ZX, LU F, LIU HY, et al. An integrated strategy by using target tissue metabolomics biomarkers as pharmacodynamic surrogate indices to screen antipyretic components of Qingkaikling injection[J]. Sci Rep, 2017, 7: 6310. doi: 10.1038/s41598-017-05812-0 [10] ZHANG F, WANG D, LI X, et al. Metabolomic study of the fever model induced by baker's yeast and the antipyretic effects of aspirin in rats using nuclear[J]. J Pharm Biomed Anal, 2013, 81/82: 168-177. doi: 10.1016/j.jpba.2013.04.005 [11] GAO XY, GUO MX, PENG L, et al. UPLC Q-TOF/MS-based metabolic profiling of urine reveals the novel antipyretic mechanisms of qingkailing injection in a rat model of yeast-induced pyrexia[J]. Evid Based Complementary Altern Med, 2013, 2013: 864747. [12] HALEY MJ, LAWRENCE CB. The blood-brain barrier after stroke: Structural studies and the role of transcytotic vesicles[J]. J Cereb Blood Flow Metab, 2017, 37(2): 456-470. doi: 10.1177/0271678X16629976 [13] WEN CC, ZHANG ML, ZHANG Y, et al. Brain metabolomics in rats after administration of ketamine[J]. Biomed Chromatogr, 2016, 30(1): 81-84. doi: 10.1002/bmc.3518 [14] SAVOLAINEN OI, SANDBERG AS, ROSS AB. A simultaneous metabolic profiling and quantitative multimetabolite metabolomic method for human plasma using gas-chromatography tandem mass spectrometry[J]. J Proteome Res, 2016, 15(1): 259-265. doi: 10.1021/acs.jproteome.5b00790 [15] DIÉMÉ B, LEFÈVRE A, NADAL-DESBARATS L, et al. Workflow methodology for rat brain metabolome exploration using NMR, LC-MS and GC-MS analytical platforms[J]. J Pharm Biomed Anal, 2017, 142: 270-278. doi: 10.1016/j.jpba.2017.03.068 [16] LEE JE, LEE YH, KIM SY, et al. Systematic biomarker discovery and coordinative validation for different primary nephrotic syndromes using gas chromatography-mass spectrometry[J]. J Chromatogr A, 2016, 1453: 105-115. doi: 10.1016/j.chroma.2016.05.058 [17] HARDEN JL, LEWIS SM, LISH SR, et al. The tryptophan metabolism enzyme L-kynureninase is a novel inflammatory factor in psoriasis and other inflammatory diseases[J]. J Allergy Clin Immunol, 2016, 137(6): 1830-1840. doi: 10.1016/j.jaci.2015.09.055 [18] ISHIWATA T, SAITO T, HASEGAWA H, et al. Changes of body temperature and extracellular serotonin level in the preoptic area and anterior hypothalamus after thermal or serotonergic pharmacological stimulation of freely moving rats[J]. Life Sci, 2004, 75(22): 2665-2675. doi: 10.1016/j.lfs.2004.04.040 [19] JIN XH, XU ZW, CAO J, et al. Proteomics analysis of human placenta reveals glutathione metabolism dysfunction as the underlying pathogenesis for preeclampsia[J]. Biochim Biophys Acta BBA Proteins Proteom, 2017, 1865(9): 1207-1214. doi: 10.1016/j.bbapap.2017.07.003 [20] GONZALEZ-RIANO C, GARCIA A, BARBAS C. Metabolomics studies in brain tissue: A review[J]. J Pharm Biomed Anal, 2016, 130: 141-168. [21] GOOD P. Does fever relieve autistic behavior by improving brain blood flow?[J]. Neuropsychol Rev, 2011, 21(1): 66-67. doi: 10.1007/s11065-011-9157-y [22] FROSINI M. Changes in CSF composition during heat stress and fever in conscious rabbits[M]//Progress in Brain Research. Amsterdam: Elsevier, 2007: 449-457. -
点击查看大图
计量
- 文章访问数: 134
- HTML全文浏览量: 32
- PDF下载量: 12
- 被引次数: 0
