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Molecular Neurobiology

, Volume 55, Issue 2, pp 1112–1122 | Cite as

Time-Dependent Lactate Production and Amino Acid Utilization in Cultured Astrocytes Under High Glucose Exposure

  • Dan Wang
  • Liangcai Zhao
  • Hong Zheng
  • Minjian Dong
  • Linlin Pan
  • Xi Zhang
  • Huajie Zhang
  • Hongchang Gao
Article

Abstract

Accumulating investigations have focused on the severity of central nervous system (CNS) complications in diabetic patients. The effects of the high glucose (HG) probably attribute to the metabolic disturbances in CNS. Astrocytes, with powerful ability of metabolic regulation, play crucial roles in physiological and pathological processes in CNS. Hence, an in-depth analysis as to metabolic alterations of astrocytes exposure to HG would facilitate to explore the underlying pathogenesis. In this study, the 1H NMR-based metabonomic approach was performed to characterize the metabolic variations of intracellular metabolites and corresponding culture media in a time-dependent manner. Our results revealed a significant elevation in lactate production and release. Four amino acids, leucine, isoleucine, methionine and tyrosine, were the most important metabolites for utilization. Also, profound disturbances of several metabolic pathways, including osmoregulation, energy metabolism, and cellular biosynthesis were observed. In that sense, the detailed information of astrocyte metabolism under HG exposure provides us a comprehensive understanding of the intrinsic metabolic disorders in CNS during hyperglycemia or diabetes.

Keywords

Astrocytes Central neural system High glucose 1H NMR Metabonomics 

Abbreviations

ANLS

Astrocyte-neuron lactate shuttle

ATP

Adenosine triphosphate

CNS

Central nervous system

DMSO

Dimethyl sulfoxide

DMEM

Dulbecco’s modified Eagle’s medium

D2O

Deuterium oxide

FBS

Fetal bovine serum

GFAP

Glial fibrillary acidic protein

GLUT

Glucose transporter

HG

High glucose

HBSS

Hank’s balanced salt solution

KEGG

Kyoto encyclopedia of genes and genomes

NMR

Nuclear magnetic resonance

MCT

Monocarboxylate transporters

NAD+

Nicotinamide adenine dinucleotide (oxidation state)

PC

Principal component

PLS-DA

Partial least squares-discriminate analysis

SMPDB

Small molecule pathway database

TCA cycle

Tricarboxylic acid cycle

TSP

TMSP-2,2,3,3-D4-sodium-3-trimethylsilylpropionate

VIP

Variable importance in the projection

Notes

Acknowledgments

This work was supported by the National Natural Science Foundation of China (Nos.: 21575105, 21175099, 21605115), Research Fund for the Doctoral Program of Higher Education of China (No. 20133321120006), Zhejiang Provincial Natural Science Foundation (Nos.:LY14H090014, LY17H160049), and Opening Project of Zhejiang Provincial Top Key Discipline of Pharmaceutical Sciences (No. YKFJ2-003).

Compliance with Ethical Standards

All animals received care, and research procedures were in accordance with the ARRIVE Guidelines [20], which were also approved by the Institutional Animal Care and Use Committee of Wenzhou Medical University.ᅟ

Conflict of Interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Dan Wang
    • 1
  • Liangcai Zhao
    • 1
  • Hong Zheng
    • 1
  • Minjian Dong
    • 1
  • Linlin Pan
    • 1
  • Xi Zhang
    • 1
  • Huajie Zhang
    • 1
  • Hongchang Gao
    • 1
  1. 1.School of Pharmaceutical SciencesWenzhou Medical UniversityWenzhouChina

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