Inhibition of transglutaminase activity reduces extracellular matrix accumulation induced by high glucose levels in proximal tubular epithelial cells

N. Skill, Timothy S. Johnson, Ian G C Coutts, Robert E. Saint, Marie Fisher, Linghong Huang, A. Meguid El Nahas, Russell J. Collighan, Martin Griffin

Research output: Contribution to journalArticle

74 Citations (Scopus)

Abstract

Diabetic nephropathy affects 30-40% of diabetics leading to end-stage kidney failure through progressive scarring and fibrosis. Previous evidence suggests that tissue transglutaminase (tTg) and its protein cross-link product ε(γ-glutamyl)lysine contribute to the expanding renal tubulointerstitial and glomerular basement membranes in this disease. Using an in vitro cell culture model of renal proximal tubular epithelial cells we determined the link between elevated glucose levels with changes in expression and activity of tTg and then, by using a highly specific site directed inhibitor of tTg (1,3-dimethyl-2[(oxopropyl)thio]imidazolium), determined the contribution of tTg to glucose-induced matrix accumulation. Exposure of cells to 36 mM glucose over 96 h caused an mRNA-dependent increase in tTg activity with a 25% increase in extracellular matrix (ECM)-associated tTg and a 150% increase in ECM ε(γ-glutamyl)lysine cross-linking. This was paralleled by an elevation in total deposited ECM resulting from higher levels of deposited collagen and fibronectin. These were associated with raised mRNA for collagens III, IV, and fibronectin. The specific site-directed inhibitor of tTg normalized both tTg activity and ECM-associated ε(γ-glutamyl)lysine. Levels of ECM per cell returned to near control levels with non-transcriptional reductions in deposited collagen and fibronectin. No changes in transforming growth factor β1 (expression or biological activity) occurred that could account for our observations, whereas incubation of tTg with collagen III indicated that cross-linking could directly increase the rate of collagen fibril/gel formation. We conclude that Tg inhibition reduces glucose-induced deposition of ECM proteins independently of changes in ECM and transforming growth factor β1 synthesis thus opening up its possible application in the treatment other fibrotic and scarring diseases where tTg has been implicated.

Original languageEnglish (US)
Pages (from-to)47754-47762
Number of pages9
JournalJournal of Biological Chemistry
Volume279
Issue number46
DOIs
StatePublished - Nov 12 2004
Externally publishedYes

Fingerprint

Transglutaminases
Extracellular Matrix
Epithelial Cells
Glucose
Collagen
Fibronectins
Lysine
Transforming Growth Factors
Cicatrix
transglutaminase 2
Kidney
Glomerular Basement Membrane
Messenger RNA
Extracellular Matrix Proteins
Level control
Diabetic Nephropathies
Bioactivity
Cell culture
Renal Insufficiency
Fibrosis

ASJC Scopus subject areas

  • Biochemistry

Cite this

Inhibition of transglutaminase activity reduces extracellular matrix accumulation induced by high glucose levels in proximal tubular epithelial cells. / Skill, N.; Johnson, Timothy S.; Coutts, Ian G C; Saint, Robert E.; Fisher, Marie; Huang, Linghong; El Nahas, A. Meguid; Collighan, Russell J.; Griffin, Martin.

In: Journal of Biological Chemistry, Vol. 279, No. 46, 12.11.2004, p. 47754-47762.

Research output: Contribution to journalArticle

Skill, N. ; Johnson, Timothy S. ; Coutts, Ian G C ; Saint, Robert E. ; Fisher, Marie ; Huang, Linghong ; El Nahas, A. Meguid ; Collighan, Russell J. ; Griffin, Martin. / Inhibition of transglutaminase activity reduces extracellular matrix accumulation induced by high glucose levels in proximal tubular epithelial cells. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 46. pp. 47754-47762.
@article{7efbd6e47f884027a0a224864bb448c5,
title = "Inhibition of transglutaminase activity reduces extracellular matrix accumulation induced by high glucose levels in proximal tubular epithelial cells",
abstract = "Diabetic nephropathy affects 30-40{\%} of diabetics leading to end-stage kidney failure through progressive scarring and fibrosis. Previous evidence suggests that tissue transglutaminase (tTg) and its protein cross-link product ε(γ-glutamyl)lysine contribute to the expanding renal tubulointerstitial and glomerular basement membranes in this disease. Using an in vitro cell culture model of renal proximal tubular epithelial cells we determined the link between elevated glucose levels with changes in expression and activity of tTg and then, by using a highly specific site directed inhibitor of tTg (1,3-dimethyl-2[(oxopropyl)thio]imidazolium), determined the contribution of tTg to glucose-induced matrix accumulation. Exposure of cells to 36 mM glucose over 96 h caused an mRNA-dependent increase in tTg activity with a 25{\%} increase in extracellular matrix (ECM)-associated tTg and a 150{\%} increase in ECM ε(γ-glutamyl)lysine cross-linking. This was paralleled by an elevation in total deposited ECM resulting from higher levels of deposited collagen and fibronectin. These were associated with raised mRNA for collagens III, IV, and fibronectin. The specific site-directed inhibitor of tTg normalized both tTg activity and ECM-associated ε(γ-glutamyl)lysine. Levels of ECM per cell returned to near control levels with non-transcriptional reductions in deposited collagen and fibronectin. No changes in transforming growth factor β1 (expression or biological activity) occurred that could account for our observations, whereas incubation of tTg with collagen III indicated that cross-linking could directly increase the rate of collagen fibril/gel formation. We conclude that Tg inhibition reduces glucose-induced deposition of ECM proteins independently of changes in ECM and transforming growth factor β1 synthesis thus opening up its possible application in the treatment other fibrotic and scarring diseases where tTg has been implicated.",
author = "N. Skill and Johnson, {Timothy S.} and Coutts, {Ian G C} and Saint, {Robert E.} and Marie Fisher and Linghong Huang and {El Nahas}, {A. Meguid} and Collighan, {Russell J.} and Martin Griffin",
year = "2004",
month = "11",
day = "12",
doi = "10.1074/jbc.M402698200",
language = "English (US)",
volume = "279",
pages = "47754--47762",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "46",

}

TY - JOUR

T1 - Inhibition of transglutaminase activity reduces extracellular matrix accumulation induced by high glucose levels in proximal tubular epithelial cells

AU - Skill, N.

AU - Johnson, Timothy S.

AU - Coutts, Ian G C

AU - Saint, Robert E.

AU - Fisher, Marie

AU - Huang, Linghong

AU - El Nahas, A. Meguid

AU - Collighan, Russell J.

AU - Griffin, Martin

PY - 2004/11/12

Y1 - 2004/11/12

N2 - Diabetic nephropathy affects 30-40% of diabetics leading to end-stage kidney failure through progressive scarring and fibrosis. Previous evidence suggests that tissue transglutaminase (tTg) and its protein cross-link product ε(γ-glutamyl)lysine contribute to the expanding renal tubulointerstitial and glomerular basement membranes in this disease. Using an in vitro cell culture model of renal proximal tubular epithelial cells we determined the link between elevated glucose levels with changes in expression and activity of tTg and then, by using a highly specific site directed inhibitor of tTg (1,3-dimethyl-2[(oxopropyl)thio]imidazolium), determined the contribution of tTg to glucose-induced matrix accumulation. Exposure of cells to 36 mM glucose over 96 h caused an mRNA-dependent increase in tTg activity with a 25% increase in extracellular matrix (ECM)-associated tTg and a 150% increase in ECM ε(γ-glutamyl)lysine cross-linking. This was paralleled by an elevation in total deposited ECM resulting from higher levels of deposited collagen and fibronectin. These were associated with raised mRNA for collagens III, IV, and fibronectin. The specific site-directed inhibitor of tTg normalized both tTg activity and ECM-associated ε(γ-glutamyl)lysine. Levels of ECM per cell returned to near control levels with non-transcriptional reductions in deposited collagen and fibronectin. No changes in transforming growth factor β1 (expression or biological activity) occurred that could account for our observations, whereas incubation of tTg with collagen III indicated that cross-linking could directly increase the rate of collagen fibril/gel formation. We conclude that Tg inhibition reduces glucose-induced deposition of ECM proteins independently of changes in ECM and transforming growth factor β1 synthesis thus opening up its possible application in the treatment other fibrotic and scarring diseases where tTg has been implicated.

AB - Diabetic nephropathy affects 30-40% of diabetics leading to end-stage kidney failure through progressive scarring and fibrosis. Previous evidence suggests that tissue transglutaminase (tTg) and its protein cross-link product ε(γ-glutamyl)lysine contribute to the expanding renal tubulointerstitial and glomerular basement membranes in this disease. Using an in vitro cell culture model of renal proximal tubular epithelial cells we determined the link between elevated glucose levels with changes in expression and activity of tTg and then, by using a highly specific site directed inhibitor of tTg (1,3-dimethyl-2[(oxopropyl)thio]imidazolium), determined the contribution of tTg to glucose-induced matrix accumulation. Exposure of cells to 36 mM glucose over 96 h caused an mRNA-dependent increase in tTg activity with a 25% increase in extracellular matrix (ECM)-associated tTg and a 150% increase in ECM ε(γ-glutamyl)lysine cross-linking. This was paralleled by an elevation in total deposited ECM resulting from higher levels of deposited collagen and fibronectin. These were associated with raised mRNA for collagens III, IV, and fibronectin. The specific site-directed inhibitor of tTg normalized both tTg activity and ECM-associated ε(γ-glutamyl)lysine. Levels of ECM per cell returned to near control levels with non-transcriptional reductions in deposited collagen and fibronectin. No changes in transforming growth factor β1 (expression or biological activity) occurred that could account for our observations, whereas incubation of tTg with collagen III indicated that cross-linking could directly increase the rate of collagen fibril/gel formation. We conclude that Tg inhibition reduces glucose-induced deposition of ECM proteins independently of changes in ECM and transforming growth factor β1 synthesis thus opening up its possible application in the treatment other fibrotic and scarring diseases where tTg has been implicated.

UR - http://www.scopus.com/inward/record.url?scp=9144241175&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=9144241175&partnerID=8YFLogxK

U2 - 10.1074/jbc.M402698200

DO - 10.1074/jbc.M402698200

M3 - Article

C2 - 15326185

AN - SCOPUS:9144241175

VL - 279

SP - 47754

EP - 47762

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 46

ER -