中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2019, Vol. 14 ›› Issue (3): 485-490.doi: 10.4103/1673-5374.245474

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

氧糖剥夺的乙醇暴露未成熟大鼠小脑颗粒细胞能量代谢受损

  

  • 出版日期:2019-03-15 发布日期:2019-03-15
  • 基金资助:

    该项目得到了罗马尼亚国家科学研究局的资助（PN-II-PT-PCCA-2011-3，No 80/2012）

Impaired energy metabolism after oxygen-glucose deprivation in immature cerebellar granular neurons exposed to ethanol

Ana Spataru 1, 2 , Diana Le Duc 3, Leon Zagrean 1 , Ana-Maria Zagrean 1   

  1. 1 Division of Physiology and Neuroscience, Carol Davila University of Medicine and Pharmacy, Bucharest, Romania
    2 King’s College Hospital, London, UK
    3 Institute of Human Genetics, University of Leipzig Hospitals and Clinics, Leipzig, Germany; Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany
  • Online:2019-03-15 Published:2019-03-15
  • Contact: Ana-Maria Zagrean, MD, PhD, ana-maria.zagrean@umfcd.ro; Diana Le Duc, MD, PhD, Gabriela-Diana.LeDuc@medizin.uni-leipzig.de.
  • Supported by:

    The project was supported by a grant of the Romanian National Authority for Scientific Research, project PN-II-PT-PCCA-2011-3, No 80/2012.

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摘要：

研究已证实乙醇对未成熟的神经系统有害，但缺血缺氧对其的影响尚有待进一步研究。为了解慢性体外乙醇暴露对未成熟大鼠小脑颗粒细胞（CGCs）能量代谢的影响以及它们对氧糖剥夺的反应，实验将原代小脑颗粒细胞培养物暴露于乙醇中。第8天，使用实时PCR定量ATP合酶基因ATP5g3的表达，然后将培养物暴露于3h的氧糖剥夺或含氧量正常的条件下。随后，通过刃天青法和ATP水平检测评估细胞代谢情况。慢性乙醇处理后ATP5g3表达、刃天青代谢和ATP水平均明显降低。氧糖剥夺暴露乙醇处理的培养物后，刃天青代谢和ATP水平进一步降低。这些结果表明，慢性乙醇暴露可能通过基因表达下调机制降低未成熟小脑颗粒细胞ATP水平，并增加了对氧糖剥夺的敏感性。

orcid: 0000-0002-4302-629X(Ana-Maria Zagrean)
           0000-0001-7289-2552(Diana Le Duc)

关键词: 细胞培养, 慢性乙醇暴露, 氧糖剥夺, 小脑颗粒细胞, 毒性, 基因表达, 细胞ATP, 细胞代谢, 代谢障碍, 细胞死亡

Abstract:

Alcohol, a widely abused drug, has deleterious effects on the immature nervous system. This study investigates the effect of chronic in vitro ethanol exposure on the metabolism of immature rat cerebellar granular cells (CGCs) and on their response to oxygen-glucose deprivation (OGD). Primary CGC cultures were exposed to ethanol (100 mM in culture medium) or to control ethanol-free medium starting day one in vitro (DIV1). At DIV8, the expression of ATP synthase gene ATP5g3 was quantified using real-time PCR, then cultures were exposed to 3 hours of OGD or normoxic conditions. Subsequently, cellular metabolism was assessed by a resazurin assay and by ATP level measurement. ATP5g3 expression was reduced by 12-fold (P = 0.03) and resazurin metabolism and ATP level were decreased to 74.4 ± 4.6% and 55.5 ± 6.9%, respectively after chronic ethanol treatment compared to control values (P < 0.01). Additionally, after OGD exposure of ethanol-treated cultures, resazurin metabolism and ATP level were decreased to 12.7 ± 1.0% and 9.0 ± 2.0% from control values (P < 0.01). These results suggest that chronic ethanol exposure reduces the cellular ATP level, possibly through a gene expression down-regulation mechanism, and increases the vulnerability to oxygen-glucose deprivation. Thus, interventions which improve metabolic function and sustain ATP-levels could attenuate ethanol-induced neuronal dysfunction and should be addressed in future studies.

Key words: cell culture, chronic ethanol exposure, oxygen-glucose deprivation, cerebellar granule cells, toxicity, gene expression, cellular ATP, cellular metabolism, metabolic impairment, cell death