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Analysis of brain metabolites by gas chromatography–mass spectrometry reveals the risk–benefit concerns of prednisone in MRL/lpr lupus mice

  • Jia Zhou
  • Feilong Lu
  • Shan Li
  • Meijuan Xie
  • Haimei Lu
  • Zhijun Xie
  • Dehong Wu
  • Shuang Wang
  • Chengping WenEmail author
  • Zheng-Hao XuEmail author
Original Article

Abstract

Objective

Neuropsychiatric systemic lupus erythematosus (NPSLE) is a common cause of disability in systemic lupus erythematosus (SLE). This study aims to investigate the metabolic changes in the hypothalamus and frontal cortex in lupus-prone MRL/lpr mice.

Methods

Metabolic changes were analyzed using gas chromatography-mass spectrometry (GC–MS).

Results

According to the principal component analysis (PCA), the metabolic profiles were different between the frontal cortex and hypothalamus, but they were comparable between MRL/lpr and MRL/MpJ mice (16 weeks of age). By OPLS-DA, eight cortical and six hypothalamic differential metabolites were identified in MRL/lpr as compared to MRL/MpJ mice. Among these differential metabolites, we found a decrease of N-acetyl-l-aspartate (NAA, a potential marker of neuronal integrity), an increase of pyruvate and a decrease of glutamate in the frontal cortex but not in the hypothalamus. Prednisone treatment (3 mg/kg from 8 weeks of age) relieved the decrease of NAA but further increased the accumulation of pyruvate in the frontal cortex, additionally affected eight enriched pathways in the hypothalamus, and led to significant imbalances between the excitation and inhibition in both the frontal cortex and hypothalamus.

Conclusion

These results suggest that the frontal cortex may be more preferentially affected than the hypothalamus in SLE. Prednisone disrupted rather than relieved metabolic abnormalities in the brain, especially in the hypothalamus, indicating that the risk–benefit balance of prednisone for SLE or NPSLE remains to be further evaluated.

Keywords

Brain Lupus Glucocorticoids Metabolomics Gas chromatography-mass spectrometry 

Abbreviations

AMP

Adenosine-monophosphate

ANOVA

Analysis of variance

EI

Electron ionization

GABA

γ-Aminobutyric acid

GC

Glucocorticoids (prednisone group)

GC–MS

Gas chromatography-mass spectrometry

HPA

Hypothalamic–pituitary–adrenal axis

IP3

Inositol triphosphates

Lpr

MRL/lpr mice (control)

MpJ

MRL/MpJ mice (normal group)

NAA

N-acetyl-L-aspartate

NPSLE

Neuropsychiatric SLE

OPLS-DA

Orthogonal partial least square-discriminant analysis

PCA

Principal component analysis

QC

Quality control

SLE

Systemic lupus erythematosus

VIP

Variable importance in projection

Notes

Acknowledgements

Z.X. would like to thank all members of Filiano Lab, Kurtzberg Lab and Marcus Center for Cellular Cures (MC3) at Duke University.

Author contributions

ZX, JZ and CP designed the study. FL, SL, MX and HL performed the experiments. JZ, FL and ZX analyzed the data. SW and ZX were involved with the interpretation of data in the manuscript. ZX wrote the manuscript in consultation with JZ and FL.

Funding

This work was funded by the Program of Zhejiang TCM Science and Technology Plan (2018ZZ007 and 2017ZA064), the National Natural Science Foundation of China (81673623 and 81873102), and partly the Foundation of the Zhejiang Chinese Medical University (ZYX2018002 and 2018ZG24), Special Scientific Project of Traditional Chinese Medicine (201507001-4) and National Natural Science Foundation of China (81873266).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Ethical approval

Experiments were approved by the Institutional Animal Care and Use Committee of Zhejiang Chinese Medical University (#ZSLL-2018-30).

Informed consent

Not applicable.

References

  1. Al Sawah S, Zhang X, Zhu B, Magder LS, Foster SA, Iikuni N, Petri M (2015) Effect of corticosteroid use by dose on the risk of developing organ damage over time in systemic lupus erythematosus-the Hopkins Lupus Cohort. Lupus Sci Med 2:e000066.  https://doi.org/10.1136/lupus-2014-000066 PubMedPubMedCentralCrossRefGoogle Scholar
  2. Alexander JJ, Zwingmann C, Quigg R (2005) MRL/lpr mice have alterations in brain metabolism as shown with [1H-13C] NMR spectroscopy. Neurochem Int 47:143–151.  https://doi.org/10.1016/j.neuint.2005.04.016 PubMedCrossRefGoogle Scholar
  3. Amissah-Arthur MB, Gordon C (2010) Contemporary treatment of systemic lupus erythematosus: an update for clinicians. Ther Adv Chronic Dis 1:163–175.  https://doi.org/10.1177/2040622310380100 PubMedPubMedCentralCrossRefGoogle Scholar
  4. Apostolopoulos D, Morand EF (2017) It hasn't gone away: the problem of glucocorticoid use in lupus remains. Rheumatology (Oxford) 56:i114–i122.  https://doi.org/10.1093/rheumatology/kew406 CrossRefGoogle Scholar
  5. Barden N (2004) Implication of the hypothalamic-pituitary-adrenal axis in the physiopathology of depression. J Psychiatry Neurosci 29:185–193PubMedPubMedCentralGoogle Scholar
  6. Cassano G et al (2007) Accrual of organ damage over time in Argentine patients with systemic lupus erythematosus: a multi-centre study. Clin Rheumatol 26:2017–2022.  https://doi.org/10.1007/s10067-007-0604-3 PubMedCrossRefGoogle Scholar
  7. Chong J et al (2018) MetaboAnalyst 4.0: towards more transparent and integrative metabolomics analysis. Nucleic Acids Res 46:W486–W494.  https://doi.org/10.1093/nar/gky310 PubMedPubMedCentralCrossRefGoogle Scholar
  8. Denburg SD, Carbotte RM, Denburg JA (1994) Corticosteroids and neuropsychological functioning in patients with systemic lupus-erythematosus. Arthritis Rheum 37:1311–1320.  https://doi.org/10.1002/art.1780370907 PubMedCrossRefGoogle Scholar
  9. Dunn WB et al (2011) Procedures for large-scale metabolic profiling of serum and plasma using gas chromatography and liquid chromatography coupled to mass spectrometry. Nat Protoc 6:1060–1083.  https://doi.org/10.1038/nprot.2011.335 PubMedCrossRefGoogle Scholar
  10. Galindo-Prieto B, Eriksson L, Trygg J (2015) Variable influence on projection (VIP) for OPLS models and its applicability in multivariate time series analysis. Chemom Intell Lab 146:297–304.  https://doi.org/10.1016/j.chemolab.2015.05.001 CrossRefGoogle Scholar
  11. Gao HX, Campbell SR, Cui MH, Zong P, Hee-Hwang J, Gulinello M, Putterman C (2009) Depression is an early disease manifestation in lupus-prone MRL/lpr mice. J Neuroimmunol 207:45–56.  https://doi.org/10.1016/j.jneuroim.2008.11.009 PubMedPubMedCentralCrossRefGoogle Scholar
  12. Gladman DD, Urowitz MB, Rahman P, Ibanez D, Tam LS (2003) Accrual of organ damage over time in patients with systemic lupus erythematosus. J Rheumatol 30:1955–1959PubMedGoogle Scholar
  13. Guidelines for referral and management of systemic lupus erythematosus in adults. American College of Rheumatology Ad Hoc Committee on Systemic Lupus Erythematosus Guidelines (1999) Arthritis Rheum 42:1785-1796 10.1002/1529-0131(199909)42:9%3c1785::AID-ANR1%3e3.0.CO;2-#Google Scholar
  14. Hanly JG (2014) Diagnosis and management of neuropsychiatric SLE. Nat Rev Rheumatol 10:338–347.  https://doi.org/10.1038/nrrheum.2014.15 PubMedCrossRefGoogle Scholar
  15. Hanly JG, Cassell K, Fisk JD (1997) Cognitive function in systemic lupus erythematosus: results of a 5-year prospective study. Arthritis Rheum 40:1542–1543.  https://doi.org/10.1002/1529-0131(199708)40:8%3c1542:AID-ART26%3e3.0.CO;2-9 PubMedCrossRefGoogle Scholar
  16. Harle P et al (2006) Increase of sympathetic outflow measured by neuropeptide Y and decrease of the hypothalamic-pituitary-adrenal axis tone in patients with systemic lupus erythematosus and rheumatoid arthritis: another example of uncoupling of response systems. Ann Rheum Dis 65:51–56.  https://doi.org/10.1136/ard.2005.038059 PubMedCrossRefGoogle Scholar
  17. Huang X, Magder LS, Petri M (2016) Predictors of incident seizure in systemic lupus erythematosus. J Rheumatol 43:565–575.  https://doi.org/10.3899/jrheum.150135 PubMedCrossRefGoogle Scholar
  18. Ivanisevic J, Siuzdak G (2015) The role of metabolomics in brain metabolism research. J Neuroimmune Pharmacol 10:391–395.  https://doi.org/10.1007/s11481-015-9621-1 PubMedPubMedCentralCrossRefGoogle Scholar
  19. Jeltsch-David H, Muller S (2014) Neuropsychiatric systemic lupus erythematosus and cognitive dysfunction: the MRL-lpr mouse strain as a model. Autoimmun Rev 13:963–973.  https://doi.org/10.1016/j.autrev.2014.08.015 PubMedCrossRefGoogle Scholar
  20. Kaul A et al (2016) Systemic lupus erythematosus. Nat Rev Dis Primers 2:16039.  https://doi.org/10.1038/nrdp.2016.39 PubMedCrossRefGoogle Scholar
  21. Keller J, Gomez R, Williams G, Lembke A, Lazzeroni L, Murphy GM Jr, Schatzberg AF (2017) HPA axis in major depression: cortisol, clinical symptomatology and genetic variation predict cognition. Mol Psychiatry 22:527–536.  https://doi.org/10.1038/mp.2016.120 PubMedCrossRefGoogle Scholar
  22. Koller MD, Templ E, Riedl M, Clodi M, Wagner O, Smolen JS, Luger A (2004) Pituitary function in patients with newly diagnosed untreated systemic lupus erythematosus. Ann Rheum Dis 63:1677–1680.  https://doi.org/10.1136/ard.2003.018325 PubMedPubMedCentralCrossRefGoogle Scholar
  23. Li Q, Wei S, Wu D, Wen C, Zhou J (2018) Urinary metabolomics study of patients with gout using gas chromatography-mass spectrometry. Biomed Res Int 2018:3461572.  https://doi.org/10.1155/2018/3461572 PubMedPubMedCentralCrossRefGoogle Scholar
  24. Lopez M, Nogueiras R, Tena-Sempere M, Dieguez C (2016) Hypothalamic AMPK: a canonical regulator of whole-body energy balance. Nat Rev Endocrinol 12:421–432.  https://doi.org/10.1038/nrendo.2016.67 PubMedCrossRefGoogle Scholar
  25. Lu F et al (2019) Limited preventive effect of prednisone on neuropsychiatric symptoms in murine systemic lupus erythematosus. Inflammopharmacology.  https://doi.org/10.1007/s10787-019-00587-4 PubMedCrossRefGoogle Scholar
  26. Monahan RC, Voorde LJJB, Steup-Beekman GM, Magro-Checa C, Huizinga TWJ, Hoekman J, Kaptein AA (2017) Neuropsychiatric symptoms in systemic lupus erythematosus: impact on quality of life. Lupus 26:1252–1259.  https://doi.org/10.1177/0961203317694262 PubMedPubMedCentralCrossRefGoogle Scholar
  27. Muller C, Binder U, Bracher F, Giera M (2017) Antifungal drug testing by combining minimal inhibitory concentration testing with target identification by gas chromatography-mass spectrometry. Nat Protoc 12:947–963PubMedCrossRefGoogle Scholar
  28. Ntziachristos V, Pleitez MA, Aime S, Brindle KM (2018) Emerging technologies to image tissue metabolism. Cell Metab.  https://doi.org/10.1016/j.cmet.2018.09.004 PubMedCrossRefGoogle Scholar
  29. Richard ML, Gilkeson G (2018) Mouse models of lupus: what they tell us and what they don't. Lupus Sci Med 5:e000199.  https://doi.org/10.1136/lupus-2016-000199 PubMedPubMedCentralCrossRefGoogle Scholar
  30. Sakic B (2018) The MRL model: a valuable tool in studies of autoimmunity-brain interactions. Methods Mol Biol 1781:259–285.  https://doi.org/10.1007/978-1-4939-7828-1_14 PubMedCrossRefGoogle Scholar
  31. Schwartz N, Stock AD, Putterman C (2019) Neuropsychiatric lupus: new mechanistic insights and future treatment directions. Nat Rev Rheumatol.  https://doi.org/10.1038/s41584-018-0156-8 PubMedCrossRefGoogle Scholar
  32. Srikumar BN et al (2018) Diminished responses to monoaminergic antidepressants but not ketamine in a mouse model for neuropsychiatric lupus. J Psychopharmacol.  https://doi.org/10.1177/0269881118812102 PubMedCrossRefGoogle Scholar
  33. Stojan G, Petri M (2017) The risk benefit ratio of glucocorticoids in SLE: have things changed over the past 40 years? Curr Treat Opt Rheumatol 3:164–172.  https://doi.org/10.1007/s40674-017-0069-8 CrossRefGoogle Scholar
  34. Suzuki S et al (2012) BDNF-dependent accumulation of palmitoleic acid in CNS neurons. Cell Mol Neurobiol 32:1367–1373.  https://doi.org/10.1007/s10571-012-9863-x PubMedCrossRefGoogle Scholar
  35. Trygg J, Wold S (2002) Orthogonal projections to latent structures (O-PLS). J Chemom 16:119–128.  https://doi.org/10.1002/cem.695 CrossRefGoogle Scholar
  36. Vo A et al (2014) Regional brain metabolism in a murine systemic lupus erythematosus model. J Cereb Blood Flow Metab 34:1315–1320.  https://doi.org/10.1038/jcbfm.2014.85 PubMedPubMedCentralCrossRefGoogle Scholar
  37. Wang W, Yang GJ, Zhang J, Chen C, Jia ZY, Li J, Xu WD (2016) Plasma, urine and ligament tissue metabolite profiling reveals potential biomarkers of ankylosing spondylitis using NMR-based metabolic profiles. Arthritis Res Ther 18:244.  https://doi.org/10.1186/s13075-016-1139-2 PubMedPubMedCentralCrossRefGoogle Scholar
  38. Wang D, Kong J, Wu J, Wang X, Lai M (2017) GC-MS-based metabolomics identifies an amino acid signature of acute ischemic stroke. Neurosci Lett 642:7–13.  https://doi.org/10.1016/j.neulet.2017.01.039 PubMedCrossRefGoogle Scholar
  39. Xu Z et al (2016) Entorhinal principal neurons mediate brain-stimulation treatments for epilepsy. EBioMedicine 14:148–160.  https://doi.org/10.1016/j.ebiom.2016.11.027 PubMedPubMedCentralCrossRefGoogle Scholar
  40. Zhou J et al (2016) Exploration of the serum metabolite signature in patients with rheumatoid arthritis using gas chromatography-mass spectrometry. J Pharm Biomed Anal 127:60–67.  https://doi.org/10.1016/j.jpba.2016.02.004 PubMedCrossRefGoogle Scholar
  41. Zhu TY, Tam LS, Lee VW, Lee KK, Li EK (2009) Systemic lupus erythematosus with neuropsychiatric manifestation incurs high disease costs: a cost-of-illness study in Hong Kong. Rheumatology (Oxford) 48:564–568.  https://doi.org/10.1093/rheumatology/kep031 CrossRefGoogle Scholar
  42. Zonana-Nacach A, Santana-Sahagun E, Jimenez-Balderas FJ, Camargo-Coronel A (2008) Prevalence and factors associated with metabolic syndrome in patients with rheumatoid arthritis and systemic lupus erythematosus. J Clin Rheumatol 14:74–77.  https://doi.org/10.1097/RHU.0b013e31816b2faa PubMedCrossRefGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Institute of TCM Clinical Basic MedicineZhejiang Chinese Medical UniversityHangzhouChina
  2. 2.The Second Affiliated Hospital of ZhejiangChinese Medical UniversityHangzhouChina
  3. 3.Epilepsy CenterThe Second Affiliated Hospital, Zhejiang University School of MedicineHangzhouChina

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