Role of 12-Lipoxygenase in the Stimulation of p38 Mitogen-Activated Protein Kinase and Collagen α5(IV) in Experimental Diabetic Nephropathy and in Glucose-Stimulated Podocytes

Shin-Wook Kang, Rama Natarajan, Asha Shahed, Cynthia C. Nast, Janine Lapage, Peter Mundel, Clifford Kashtan, Sharon G. Adler

Research output: Contribution to journalArticle

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Abstract

The 12-lipoxygenase (12-LO) pathway of arachidonic acid metabolism is implicated in extracellular matrix (ECM) synthesis, but its role in podocytes has not been studied. This study tested whether 12-LO induction by diabetes or by high glucose (HG) in cultured podocytes alters glomerular basement membrane by activating signal transduction pathways culminating in ECM synthesis. Sprague-Dawley rats received an injection of diluent (control [C]) or streptozotocin 65 mg/kg (DM) and were killed at 1 or 4 mo. Glomerular 12-LO mRNA and protein levels were higher in DM than in C glomeruli at 1 and 4 mo, and 12-LO localized predominantly in podocytes. Glomerular p38 mRNA and protein were higher in DM at months 1 and 4, but phosphop38 mitogen-activated protein (MAPK) was increased only at month 1. Glomerular collagen α 5(IV)/glutaraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA ratio was increased in DM at month 1 but not at month 4, whereas collagen α5(IV) protein was higher at both 1 and 4 mo. Mouse podocytes were cultured in media with 25 mM glucose (HG) with or without the 12-LO inhibitor cinnamyl-3,4-dihydroxy-cyanocinnamate (CDC) or with 5.5 mM glucose + 19.5 mM mannitol (low glucose [LG+M]) for 10 d at 37°C. 12-LO mRNA and protein levels were higher in HG than in LG+M as was the p38 MAPK/GAPDH mRNA ratio. Phospho-p38 MAPK protein but not total p38 MAPK was higher in HG compared with LG+M. Collagen α5(IV)/ GAPDH mRNA ratio and protein were higher in HG than in LG+M. 12-LO inhibition by CDC decreased HG-induced phospho-p38 MAPK and the phospho-p38/total p38 MAPK ratio, collagen α5(IV)/GAPDH mRNA ratio, and collagen α5(IV) protein expression. In summary, diabetes in vivo and exposure of podocytes to HG in vitro stimulated 12-LO, p38 MAPK, and collagen α5(IV) mRNA and (activated) protein. 12-LO inhibition by CDC diminished the expression of podocyte phospho-p38 MAPK and collagen α5(IV) mRNA and protein. These findings implicate 12-LO and the p38 MAPK signaling pathway in the mediation of ECM synthesis by podocytes in diabetes.

Original languageEnglish
Pages (from-to)3178-3187
Number of pages10
JournalJournal of the American Society of Nephrology
Volume14
Issue number12
DOIs
Publication statusPublished - 2003 Dec 1

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Arachidonate 12-Lipoxygenase
Podocytes
Diabetic Nephropathies
p38 Mitogen-Activated Protein Kinases
Collagen
Glucose
Messenger RNA
Glutaral
Proteins
Oxidoreductases
Phosphates
Extracellular Matrix
Lipoxygenase Inhibitors
Glomerular Basement Membrane
Mannitol
Streptozocin
Mitogens
Sprague Dawley Rats
Signal Transduction

All Science Journal Classification (ASJC) codes

  • Nephrology

Cite this

Kang, Shin-Wook ; Natarajan, Rama ; Shahed, Asha ; Nast, Cynthia C. ; Lapage, Janine ; Mundel, Peter ; Kashtan, Clifford ; Adler, Sharon G. / Role of 12-Lipoxygenase in the Stimulation of p38 Mitogen-Activated Protein Kinase and Collagen α5(IV) in Experimental Diabetic Nephropathy and in Glucose-Stimulated Podocytes. In: Journal of the American Society of Nephrology. 2003 ; Vol. 14, No. 12. pp. 3178-3187.
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Role of 12-Lipoxygenase in the Stimulation of p38 Mitogen-Activated Protein Kinase and Collagen α5(IV) in Experimental Diabetic Nephropathy and in Glucose-Stimulated Podocytes. / Kang, Shin-Wook; Natarajan, Rama; Shahed, Asha; Nast, Cynthia C.; Lapage, Janine; Mundel, Peter; Kashtan, Clifford; Adler, Sharon G.

In: Journal of the American Society of Nephrology, Vol. 14, No. 12, 01.12.2003, p. 3178-3187.

Research output: Contribution to journalArticle

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T1 - Role of 12-Lipoxygenase in the Stimulation of p38 Mitogen-Activated Protein Kinase and Collagen α5(IV) in Experimental Diabetic Nephropathy and in Glucose-Stimulated Podocytes

AU - Kang, Shin-Wook

AU - Natarajan, Rama

AU - Shahed, Asha

AU - Nast, Cynthia C.

AU - Lapage, Janine

AU - Mundel, Peter

AU - Kashtan, Clifford

AU - Adler, Sharon G.

PY - 2003/12/1

Y1 - 2003/12/1

N2 - The 12-lipoxygenase (12-LO) pathway of arachidonic acid metabolism is implicated in extracellular matrix (ECM) synthesis, but its role in podocytes has not been studied. This study tested whether 12-LO induction by diabetes or by high glucose (HG) in cultured podocytes alters glomerular basement membrane by activating signal transduction pathways culminating in ECM synthesis. Sprague-Dawley rats received an injection of diluent (control [C]) or streptozotocin 65 mg/kg (DM) and were killed at 1 or 4 mo. Glomerular 12-LO mRNA and protein levels were higher in DM than in C glomeruli at 1 and 4 mo, and 12-LO localized predominantly in podocytes. Glomerular p38 mRNA and protein were higher in DM at months 1 and 4, but phosphop38 mitogen-activated protein (MAPK) was increased only at month 1. Glomerular collagen α 5(IV)/glutaraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA ratio was increased in DM at month 1 but not at month 4, whereas collagen α5(IV) protein was higher at both 1 and 4 mo. Mouse podocytes were cultured in media with 25 mM glucose (HG) with or without the 12-LO inhibitor cinnamyl-3,4-dihydroxy-cyanocinnamate (CDC) or with 5.5 mM glucose + 19.5 mM mannitol (low glucose [LG+M]) for 10 d at 37°C. 12-LO mRNA and protein levels were higher in HG than in LG+M as was the p38 MAPK/GAPDH mRNA ratio. Phospho-p38 MAPK protein but not total p38 MAPK was higher in HG compared with LG+M. Collagen α5(IV)/ GAPDH mRNA ratio and protein were higher in HG than in LG+M. 12-LO inhibition by CDC decreased HG-induced phospho-p38 MAPK and the phospho-p38/total p38 MAPK ratio, collagen α5(IV)/GAPDH mRNA ratio, and collagen α5(IV) protein expression. In summary, diabetes in vivo and exposure of podocytes to HG in vitro stimulated 12-LO, p38 MAPK, and collagen α5(IV) mRNA and (activated) protein. 12-LO inhibition by CDC diminished the expression of podocyte phospho-p38 MAPK and collagen α5(IV) mRNA and protein. These findings implicate 12-LO and the p38 MAPK signaling pathway in the mediation of ECM synthesis by podocytes in diabetes.

AB - The 12-lipoxygenase (12-LO) pathway of arachidonic acid metabolism is implicated in extracellular matrix (ECM) synthesis, but its role in podocytes has not been studied. This study tested whether 12-LO induction by diabetes or by high glucose (HG) in cultured podocytes alters glomerular basement membrane by activating signal transduction pathways culminating in ECM synthesis. Sprague-Dawley rats received an injection of diluent (control [C]) or streptozotocin 65 mg/kg (DM) and were killed at 1 or 4 mo. Glomerular 12-LO mRNA and protein levels were higher in DM than in C glomeruli at 1 and 4 mo, and 12-LO localized predominantly in podocytes. Glomerular p38 mRNA and protein were higher in DM at months 1 and 4, but phosphop38 mitogen-activated protein (MAPK) was increased only at month 1. Glomerular collagen α 5(IV)/glutaraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA ratio was increased in DM at month 1 but not at month 4, whereas collagen α5(IV) protein was higher at both 1 and 4 mo. Mouse podocytes were cultured in media with 25 mM glucose (HG) with or without the 12-LO inhibitor cinnamyl-3,4-dihydroxy-cyanocinnamate (CDC) or with 5.5 mM glucose + 19.5 mM mannitol (low glucose [LG+M]) for 10 d at 37°C. 12-LO mRNA and protein levels were higher in HG than in LG+M as was the p38 MAPK/GAPDH mRNA ratio. Phospho-p38 MAPK protein but not total p38 MAPK was higher in HG compared with LG+M. Collagen α5(IV)/ GAPDH mRNA ratio and protein were higher in HG than in LG+M. 12-LO inhibition by CDC decreased HG-induced phospho-p38 MAPK and the phospho-p38/total p38 MAPK ratio, collagen α5(IV)/GAPDH mRNA ratio, and collagen α5(IV) protein expression. In summary, diabetes in vivo and exposure of podocytes to HG in vitro stimulated 12-LO, p38 MAPK, and collagen α5(IV) mRNA and (activated) protein. 12-LO inhibition by CDC diminished the expression of podocyte phospho-p38 MAPK and collagen α5(IV) mRNA and protein. These findings implicate 12-LO and the p38 MAPK signaling pathway in the mediation of ECM synthesis by podocytes in diabetes.

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