Acute glucose deprivation leads to apoptosis in a cell model of acute diabetic neuropathy

Hiroyuki Honma, Jewel L. Podratz, Anthony John Windebank

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Objective: Our aims were to better understand the mechanisms underlying peripheral neuropathy with diabetes mellitus and to test the hypothesis that acute lowering of glucose levels induces apoptosis in hypoxic neurons. Methods: We used rat dissociated dorsal root ganglion (DRG) neurons incubated in a medium high in glucose concentration (700 mg%) and room air (PO2 150 torr). After 5 days, DRG neurons were placed in hypoxic conditions (PO 2 7.6 torr) with a normal-glucose (100 mg%) or high-glucose (700 mg%) medium containing 3 of 100 ng/mL of nerve growth factor. Acute lowering of glucose levels under hypoxic conditions led to apoptosis of DRG neurons. Apoptosis was demonstrated by bis-benzimide staining for nuclear fragmentation, electron microscopy, DNA laddering, and TUNEL staining. Caspase 3 immunocytochemistry and inhibition of neuronal death by the caspase inhibitor z-VAD-fmk (100 μM) confirmed that death was apoptotic. Results: Hypoxia-induced death was decreased when DRG neurons were maintained in high-glucose medium, suggesting that high levels of substrate protected against hypoxia. Apoptosis was completely prevented by increasing the concentration of nerve growth factor from 3 to 100 ng/mL and was partially prevented by the addition of the antioxidant α-lipoic acid (500 μM). Conclusions: This model provides a novel means for studying the pathogenesis and treatment of early stages of diabetic neuropathy.

Original languageEnglish (US)
Pages (from-to)65-74
Number of pages10
JournalJournal of the Peripheral Nervous System
Volume8
Issue number2
DOIs
StatePublished - Jun 2003

Fingerprint

Diabetic Neuropathies
Spinal Ganglia
Apoptosis
Glucose
Neurons
Nerve Growth Factor
Staining and Labeling
Thioctic Acid
Caspase Inhibitors
In Situ Nick-End Labeling
Peripheral Nervous System Diseases
Caspase 3
Electron Microscopy
Diabetes Mellitus
Antioxidants
Immunohistochemistry
Air
DNA

Keywords

  • Apoptosis
  • Diabetic neuropathy
  • Glucose

ASJC Scopus subject areas

  • Clinical Neurology
  • Neuroscience(all)

Cite this

Acute glucose deprivation leads to apoptosis in a cell model of acute diabetic neuropathy. / Honma, Hiroyuki; Podratz, Jewel L.; Windebank, Anthony John.

In: Journal of the Peripheral Nervous System, Vol. 8, No. 2, 06.2003, p. 65-74.

Research output: Contribution to journalArticle

@article{d1794e744182483d96e5b4cac1b5622b,
title = "Acute glucose deprivation leads to apoptosis in a cell model of acute diabetic neuropathy",
abstract = "Objective: Our aims were to better understand the mechanisms underlying peripheral neuropathy with diabetes mellitus and to test the hypothesis that acute lowering of glucose levels induces apoptosis in hypoxic neurons. Methods: We used rat dissociated dorsal root ganglion (DRG) neurons incubated in a medium high in glucose concentration (700 mg{\%}) and room air (PO2 150 torr). After 5 days, DRG neurons were placed in hypoxic conditions (PO 2 7.6 torr) with a normal-glucose (100 mg{\%}) or high-glucose (700 mg{\%}) medium containing 3 of 100 ng/mL of nerve growth factor. Acute lowering of glucose levels under hypoxic conditions led to apoptosis of DRG neurons. Apoptosis was demonstrated by bis-benzimide staining for nuclear fragmentation, electron microscopy, DNA laddering, and TUNEL staining. Caspase 3 immunocytochemistry and inhibition of neuronal death by the caspase inhibitor z-VAD-fmk (100 μM) confirmed that death was apoptotic. Results: Hypoxia-induced death was decreased when DRG neurons were maintained in high-glucose medium, suggesting that high levels of substrate protected against hypoxia. Apoptosis was completely prevented by increasing the concentration of nerve growth factor from 3 to 100 ng/mL and was partially prevented by the addition of the antioxidant α-lipoic acid (500 μM). Conclusions: This model provides a novel means for studying the pathogenesis and treatment of early stages of diabetic neuropathy.",
keywords = "Apoptosis, Diabetic neuropathy, Glucose",
author = "Hiroyuki Honma and Podratz, {Jewel L.} and Windebank, {Anthony John}",
year = "2003",
month = "6",
doi = "10.1046/j.1529-8027.2003.03009.x",
language = "English (US)",
volume = "8",
pages = "65--74",
journal = "Journal of the Peripheral Nervous System",
issn = "1085-9489",
publisher = "Wiley-Blackwell",
number = "2",

}

TY - JOUR

T1 - Acute glucose deprivation leads to apoptosis in a cell model of acute diabetic neuropathy

AU - Honma, Hiroyuki

AU - Podratz, Jewel L.

AU - Windebank, Anthony John

PY - 2003/6

Y1 - 2003/6

N2 - Objective: Our aims were to better understand the mechanisms underlying peripheral neuropathy with diabetes mellitus and to test the hypothesis that acute lowering of glucose levels induces apoptosis in hypoxic neurons. Methods: We used rat dissociated dorsal root ganglion (DRG) neurons incubated in a medium high in glucose concentration (700 mg%) and room air (PO2 150 torr). After 5 days, DRG neurons were placed in hypoxic conditions (PO 2 7.6 torr) with a normal-glucose (100 mg%) or high-glucose (700 mg%) medium containing 3 of 100 ng/mL of nerve growth factor. Acute lowering of glucose levels under hypoxic conditions led to apoptosis of DRG neurons. Apoptosis was demonstrated by bis-benzimide staining for nuclear fragmentation, electron microscopy, DNA laddering, and TUNEL staining. Caspase 3 immunocytochemistry and inhibition of neuronal death by the caspase inhibitor z-VAD-fmk (100 μM) confirmed that death was apoptotic. Results: Hypoxia-induced death was decreased when DRG neurons were maintained in high-glucose medium, suggesting that high levels of substrate protected against hypoxia. Apoptosis was completely prevented by increasing the concentration of nerve growth factor from 3 to 100 ng/mL and was partially prevented by the addition of the antioxidant α-lipoic acid (500 μM). Conclusions: This model provides a novel means for studying the pathogenesis and treatment of early stages of diabetic neuropathy.

AB - Objective: Our aims were to better understand the mechanisms underlying peripheral neuropathy with diabetes mellitus and to test the hypothesis that acute lowering of glucose levels induces apoptosis in hypoxic neurons. Methods: We used rat dissociated dorsal root ganglion (DRG) neurons incubated in a medium high in glucose concentration (700 mg%) and room air (PO2 150 torr). After 5 days, DRG neurons were placed in hypoxic conditions (PO 2 7.6 torr) with a normal-glucose (100 mg%) or high-glucose (700 mg%) medium containing 3 of 100 ng/mL of nerve growth factor. Acute lowering of glucose levels under hypoxic conditions led to apoptosis of DRG neurons. Apoptosis was demonstrated by bis-benzimide staining for nuclear fragmentation, electron microscopy, DNA laddering, and TUNEL staining. Caspase 3 immunocytochemistry and inhibition of neuronal death by the caspase inhibitor z-VAD-fmk (100 μM) confirmed that death was apoptotic. Results: Hypoxia-induced death was decreased when DRG neurons were maintained in high-glucose medium, suggesting that high levels of substrate protected against hypoxia. Apoptosis was completely prevented by increasing the concentration of nerve growth factor from 3 to 100 ng/mL and was partially prevented by the addition of the antioxidant α-lipoic acid (500 μM). Conclusions: This model provides a novel means for studying the pathogenesis and treatment of early stages of diabetic neuropathy.

KW - Apoptosis

KW - Diabetic neuropathy

KW - Glucose

UR - http://www.scopus.com/inward/record.url?scp=0041784076&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0041784076&partnerID=8YFLogxK

U2 - 10.1046/j.1529-8027.2003.03009.x

DO - 10.1046/j.1529-8027.2003.03009.x

M3 - Article

C2 - 12795710

AN - SCOPUS:0041784076

VL - 8

SP - 65

EP - 74

JO - Journal of the Peripheral Nervous System

JF - Journal of the Peripheral Nervous System

SN - 1085-9489

IS - 2

ER -