Gene expression profiling of diabetic and galactosaemic cataractous rat lens by microarray analysis

E. Kubo, Dhirendra P Singh, Y. Akagi

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Aims/hypothesis: Osmotic and oxidative stress is associated with the progression and advancement of diabetic cataract. In the present study, we used a cDNA microarray method to analyse gene expression patterns in streptozotocin-induced diabetic rats and galactose-fed cataractous lenses. In addition, we investigated the regulation and interaction(s) of anti-oxidant protein 2 and lens epithelium-derived growth factor in these models. Methods: To identify differential gene expression patterns, one group of Sprague-Dawley rats was made diabetic with streptozotocin and a second group was made galactosaemic. Total RNA was extracted from the lenses of both groups and their controls. Labelled cDNA was hybridised to Atlas Rat Arrays. Changes in gene expression level were analysed. Real-time PCR and western analysis were used to validate the microarray results. Results: The expression of 31 genes was significantly modulated in hyperglycaemic lenses compared with galactosaemic lenses. Notably, transcript and protein levels of B-cell leukaemia/lymphoma protein 2 and nuclear factor-κB were significantly elevated in rat lenses at 4 weeks after injection of streptozotocin. At a later stage, mRNA and protein levels of TGF-β were elevated. However, levels of mRNA for IGF-1, lens epithelium-derived growth factor and anti-oxidant protein 2 were diminished in streptozotocin-induced diabetic cataract. Conclusions/interpretations: These results provide evidence that progression of sugar cataract involves oxidative- and TGF-β-mediated signalling. These pathways may promote abnormal gene expression in the hyperglycaemic and galactosaemic states and thus may contribute to the symptoms associated with these conditions. Since oxidative stress seems to be a major event in cataract formation, supply of anti-oxidant may postpone the progression of such disorders.

Original languageEnglish (US)
Pages (from-to)790-798
Number of pages9
JournalDiabetologia
Volume48
Issue number4
DOIs
StatePublished - Apr 1 2005

Fingerprint

Gene Expression Profiling
Microarray Analysis
Lenses
Streptozocin
Cataract
Oxidants
Gene Expression
Oxidative Stress
B-Cell Leukemia
Messenger RNA
Proteins
Atlases
Osmotic Pressure
B-Cell Lymphoma
Nuclear Proteins
Oligonucleotide Array Sequence Analysis
Galactose
Insulin-Like Growth Factor I
Sprague Dawley Rats
Real-Time Polymerase Chain Reaction

Keywords

  • Diabetic cataract
  • Galactose cataract
  • Oxidative stress
  • Polyol pathway
  • cDNA microarray

ASJC Scopus subject areas

  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism

Cite this

Gene expression profiling of diabetic and galactosaemic cataractous rat lens by microarray analysis. / Kubo, E.; Singh, Dhirendra P; Akagi, Y.

In: Diabetologia, Vol. 48, No. 4, 01.04.2005, p. 790-798.

Research output: Contribution to journalArticle

@article{f3120c1fa72a4c349dd146636fce3978,
title = "Gene expression profiling of diabetic and galactosaemic cataractous rat lens by microarray analysis",
abstract = "Aims/hypothesis: Osmotic and oxidative stress is associated with the progression and advancement of diabetic cataract. In the present study, we used a cDNA microarray method to analyse gene expression patterns in streptozotocin-induced diabetic rats and galactose-fed cataractous lenses. In addition, we investigated the regulation and interaction(s) of anti-oxidant protein 2 and lens epithelium-derived growth factor in these models. Methods: To identify differential gene expression patterns, one group of Sprague-Dawley rats was made diabetic with streptozotocin and a second group was made galactosaemic. Total RNA was extracted from the lenses of both groups and their controls. Labelled cDNA was hybridised to Atlas Rat Arrays. Changes in gene expression level were analysed. Real-time PCR and western analysis were used to validate the microarray results. Results: The expression of 31 genes was significantly modulated in hyperglycaemic lenses compared with galactosaemic lenses. Notably, transcript and protein levels of B-cell leukaemia/lymphoma protein 2 and nuclear factor-κB were significantly elevated in rat lenses at 4 weeks after injection of streptozotocin. At a later stage, mRNA and protein levels of TGF-β were elevated. However, levels of mRNA for IGF-1, lens epithelium-derived growth factor and anti-oxidant protein 2 were diminished in streptozotocin-induced diabetic cataract. Conclusions/interpretations: These results provide evidence that progression of sugar cataract involves oxidative- and TGF-β-mediated signalling. These pathways may promote abnormal gene expression in the hyperglycaemic and galactosaemic states and thus may contribute to the symptoms associated with these conditions. Since oxidative stress seems to be a major event in cataract formation, supply of anti-oxidant may postpone the progression of such disorders.",
keywords = "Diabetic cataract, Galactose cataract, Oxidative stress, Polyol pathway, cDNA microarray",
author = "E. Kubo and Singh, {Dhirendra P} and Y. Akagi",
year = "2005",
month = "4",
day = "1",
doi = "10.1007/s00125-005-1687-5",
language = "English (US)",
volume = "48",
pages = "790--798",
journal = "Diabetologia",
issn = "0012-186X",
publisher = "Springer Verlag",
number = "4",

}

TY - JOUR

T1 - Gene expression profiling of diabetic and galactosaemic cataractous rat lens by microarray analysis

AU - Kubo, E.

AU - Singh, Dhirendra P

AU - Akagi, Y.

PY - 2005/4/1

Y1 - 2005/4/1

N2 - Aims/hypothesis: Osmotic and oxidative stress is associated with the progression and advancement of diabetic cataract. In the present study, we used a cDNA microarray method to analyse gene expression patterns in streptozotocin-induced diabetic rats and galactose-fed cataractous lenses. In addition, we investigated the regulation and interaction(s) of anti-oxidant protein 2 and lens epithelium-derived growth factor in these models. Methods: To identify differential gene expression patterns, one group of Sprague-Dawley rats was made diabetic with streptozotocin and a second group was made galactosaemic. Total RNA was extracted from the lenses of both groups and their controls. Labelled cDNA was hybridised to Atlas Rat Arrays. Changes in gene expression level were analysed. Real-time PCR and western analysis were used to validate the microarray results. Results: The expression of 31 genes was significantly modulated in hyperglycaemic lenses compared with galactosaemic lenses. Notably, transcript and protein levels of B-cell leukaemia/lymphoma protein 2 and nuclear factor-κB were significantly elevated in rat lenses at 4 weeks after injection of streptozotocin. At a later stage, mRNA and protein levels of TGF-β were elevated. However, levels of mRNA for IGF-1, lens epithelium-derived growth factor and anti-oxidant protein 2 were diminished in streptozotocin-induced diabetic cataract. Conclusions/interpretations: These results provide evidence that progression of sugar cataract involves oxidative- and TGF-β-mediated signalling. These pathways may promote abnormal gene expression in the hyperglycaemic and galactosaemic states and thus may contribute to the symptoms associated with these conditions. Since oxidative stress seems to be a major event in cataract formation, supply of anti-oxidant may postpone the progression of such disorders.

AB - Aims/hypothesis: Osmotic and oxidative stress is associated with the progression and advancement of diabetic cataract. In the present study, we used a cDNA microarray method to analyse gene expression patterns in streptozotocin-induced diabetic rats and galactose-fed cataractous lenses. In addition, we investigated the regulation and interaction(s) of anti-oxidant protein 2 and lens epithelium-derived growth factor in these models. Methods: To identify differential gene expression patterns, one group of Sprague-Dawley rats was made diabetic with streptozotocin and a second group was made galactosaemic. Total RNA was extracted from the lenses of both groups and their controls. Labelled cDNA was hybridised to Atlas Rat Arrays. Changes in gene expression level were analysed. Real-time PCR and western analysis were used to validate the microarray results. Results: The expression of 31 genes was significantly modulated in hyperglycaemic lenses compared with galactosaemic lenses. Notably, transcript and protein levels of B-cell leukaemia/lymphoma protein 2 and nuclear factor-κB were significantly elevated in rat lenses at 4 weeks after injection of streptozotocin. At a later stage, mRNA and protein levels of TGF-β were elevated. However, levels of mRNA for IGF-1, lens epithelium-derived growth factor and anti-oxidant protein 2 were diminished in streptozotocin-induced diabetic cataract. Conclusions/interpretations: These results provide evidence that progression of sugar cataract involves oxidative- and TGF-β-mediated signalling. These pathways may promote abnormal gene expression in the hyperglycaemic and galactosaemic states and thus may contribute to the symptoms associated with these conditions. Since oxidative stress seems to be a major event in cataract formation, supply of anti-oxidant may postpone the progression of such disorders.

KW - Diabetic cataract

KW - Galactose cataract

KW - Oxidative stress

KW - Polyol pathway

KW - cDNA microarray

UR - http://www.scopus.com/inward/record.url?scp=17844373867&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=17844373867&partnerID=8YFLogxK

U2 - 10.1007/s00125-005-1687-5

DO - 10.1007/s00125-005-1687-5

M3 - Article

VL - 48

SP - 790

EP - 798

JO - Diabetologia

JF - Diabetologia

SN - 0012-186X

IS - 4

ER -