12-Lipoxygenase is increased in glucose-stimulated mesangial cells and in experimental diabetic nephropathy

Shin Wook Kang, Sharon G. Adler, Cynthia C. Nast, Janine LaPage, Jia Li Gu, Jerry L. Nadler, Rama Natarajan

Research output: Contribution to journalArticle

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Abstract

Background. Arachidonic acid-derived 12-lipoxygenase (12-LO) products have potent growth and chemotactic properties. The present studies examined whether 12-LO and fibronectin are induced in cultured rat mesangial cells (MCs) exposed to high glucose and whether they are expressed in experimental diabetic nephropathy. Methods. To determine the effect of high glucose on MC 12-LO mRNA and protein expression, rat MCs were incubated with RPMI medium containing 100 (NG) or 450 mg/dL glucose (HG). For animal studies, rats were injected with diluent (control) or streptozotocin. The latter were left untreated (DM) or treated with insulin (DM + I). At sacrifice after four months, GAPDH, 12-LO, and fibronectin mRNA were measured by competitive reverse transcription-polymerase chain reaction (RT-PCR) in microdissected glomeruli (G). Renal sections were semiquantitatively scored (0 to 4+) for diabetic changes and for 12-LO and fibronectin by immunohistochemistry. Results. 12-LO mRNA expression in MC exposed to HG (12.71 ± 1.17 attm/μL) and DM G (1.78 ± 0.65 × 10-3 attm/glomerulus) was significantly higher than those of MCs in NG media (6.71 ± 0.78 attm/μL) and control G (0.34 ± 0.12 × 10-3 attm/glomerulus, P < 0.005), respectively. Western blot revealed a 1.7- and a 2.8-fold increase in MC and G 12-LO protein expression, respectively (P < 0.05). The immunohistochemistry score for G 12-LO and diabetic nephropathy score was significantly greater in DM and DM + I than controls. MC and G GAPDH mRNA remained unchanged. Conclusions. In MCs exposed to HG and in diabetic rat glomeruli, increments in 12-LO mRNA and protein are associated with changes modeling diabetic nephropathy. These findings suggest a role for the 12-LO pathway in the pathogenesis of diabetic nephropathy.

Original languageEnglish
Pages (from-to)1354-1362
Number of pages9
JournalKidney International
Volume59
Issue number4
DOIs
Publication statusPublished - 2001 Jan 1

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Arachidonate 12-Lipoxygenase
Mesangial Cells
Diabetic Nephropathies
Glucose
Messenger RNA
Fibronectins
Immunohistochemistry
Proteins
Streptozocin
Reverse Transcription
Western Blotting

All Science Journal Classification (ASJC) codes

  • Nephrology

Cite this

Kang, Shin Wook ; Adler, Sharon G. ; Nast, Cynthia C. ; LaPage, Janine ; Gu, Jia Li ; Nadler, Jerry L. ; Natarajan, Rama. / 12-Lipoxygenase is increased in glucose-stimulated mesangial cells and in experimental diabetic nephropathy. In: Kidney International. 2001 ; Vol. 59, No. 4. pp. 1354-1362.
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title = "12-Lipoxygenase is increased in glucose-stimulated mesangial cells and in experimental diabetic nephropathy",
abstract = "Background. Arachidonic acid-derived 12-lipoxygenase (12-LO) products have potent growth and chemotactic properties. The present studies examined whether 12-LO and fibronectin are induced in cultured rat mesangial cells (MCs) exposed to high glucose and whether they are expressed in experimental diabetic nephropathy. Methods. To determine the effect of high glucose on MC 12-LO mRNA and protein expression, rat MCs were incubated with RPMI medium containing 100 (NG) or 450 mg/dL glucose (HG). For animal studies, rats were injected with diluent (control) or streptozotocin. The latter were left untreated (DM) or treated with insulin (DM + I). At sacrifice after four months, GAPDH, 12-LO, and fibronectin mRNA were measured by competitive reverse transcription-polymerase chain reaction (RT-PCR) in microdissected glomeruli (G). Renal sections were semiquantitatively scored (0 to 4+) for diabetic changes and for 12-LO and fibronectin by immunohistochemistry. Results. 12-LO mRNA expression in MC exposed to HG (12.71 ± 1.17 attm/μL) and DM G (1.78 ± 0.65 × 10-3 attm/glomerulus) was significantly higher than those of MCs in NG media (6.71 ± 0.78 attm/μL) and control G (0.34 ± 0.12 × 10-3 attm/glomerulus, P < 0.005), respectively. Western blot revealed a 1.7- and a 2.8-fold increase in MC and G 12-LO protein expression, respectively (P < 0.05). The immunohistochemistry score for G 12-LO and diabetic nephropathy score was significantly greater in DM and DM + I than controls. MC and G GAPDH mRNA remained unchanged. Conclusions. In MCs exposed to HG and in diabetic rat glomeruli, increments in 12-LO mRNA and protein are associated with changes modeling diabetic nephropathy. These findings suggest a role for the 12-LO pathway in the pathogenesis of diabetic nephropathy.",
author = "Kang, {Shin Wook} and Adler, {Sharon G.} and Nast, {Cynthia C.} and Janine LaPage and Gu, {Jia Li} and Nadler, {Jerry L.} and Rama Natarajan",
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12-Lipoxygenase is increased in glucose-stimulated mesangial cells and in experimental diabetic nephropathy. / Kang, Shin Wook; Adler, Sharon G.; Nast, Cynthia C.; LaPage, Janine; Gu, Jia Li; Nadler, Jerry L.; Natarajan, Rama.

In: Kidney International, Vol. 59, No. 4, 01.01.2001, p. 1354-1362.

Research output: Contribution to journalArticle

TY - JOUR

T1 - 12-Lipoxygenase is increased in glucose-stimulated mesangial cells and in experimental diabetic nephropathy

AU - Kang, Shin Wook

AU - Adler, Sharon G.

AU - Nast, Cynthia C.

AU - LaPage, Janine

AU - Gu, Jia Li

AU - Nadler, Jerry L.

AU - Natarajan, Rama

PY - 2001/1/1

Y1 - 2001/1/1

N2 - Background. Arachidonic acid-derived 12-lipoxygenase (12-LO) products have potent growth and chemotactic properties. The present studies examined whether 12-LO and fibronectin are induced in cultured rat mesangial cells (MCs) exposed to high glucose and whether they are expressed in experimental diabetic nephropathy. Methods. To determine the effect of high glucose on MC 12-LO mRNA and protein expression, rat MCs were incubated with RPMI medium containing 100 (NG) or 450 mg/dL glucose (HG). For animal studies, rats were injected with diluent (control) or streptozotocin. The latter were left untreated (DM) or treated with insulin (DM + I). At sacrifice after four months, GAPDH, 12-LO, and fibronectin mRNA were measured by competitive reverse transcription-polymerase chain reaction (RT-PCR) in microdissected glomeruli (G). Renal sections were semiquantitatively scored (0 to 4+) for diabetic changes and for 12-LO and fibronectin by immunohistochemistry. Results. 12-LO mRNA expression in MC exposed to HG (12.71 ± 1.17 attm/μL) and DM G (1.78 ± 0.65 × 10-3 attm/glomerulus) was significantly higher than those of MCs in NG media (6.71 ± 0.78 attm/μL) and control G (0.34 ± 0.12 × 10-3 attm/glomerulus, P < 0.005), respectively. Western blot revealed a 1.7- and a 2.8-fold increase in MC and G 12-LO protein expression, respectively (P < 0.05). The immunohistochemistry score for G 12-LO and diabetic nephropathy score was significantly greater in DM and DM + I than controls. MC and G GAPDH mRNA remained unchanged. Conclusions. In MCs exposed to HG and in diabetic rat glomeruli, increments in 12-LO mRNA and protein are associated with changes modeling diabetic nephropathy. These findings suggest a role for the 12-LO pathway in the pathogenesis of diabetic nephropathy.

AB - Background. Arachidonic acid-derived 12-lipoxygenase (12-LO) products have potent growth and chemotactic properties. The present studies examined whether 12-LO and fibronectin are induced in cultured rat mesangial cells (MCs) exposed to high glucose and whether they are expressed in experimental diabetic nephropathy. Methods. To determine the effect of high glucose on MC 12-LO mRNA and protein expression, rat MCs were incubated with RPMI medium containing 100 (NG) or 450 mg/dL glucose (HG). For animal studies, rats were injected with diluent (control) or streptozotocin. The latter were left untreated (DM) or treated with insulin (DM + I). At sacrifice after four months, GAPDH, 12-LO, and fibronectin mRNA were measured by competitive reverse transcription-polymerase chain reaction (RT-PCR) in microdissected glomeruli (G). Renal sections were semiquantitatively scored (0 to 4+) for diabetic changes and for 12-LO and fibronectin by immunohistochemistry. Results. 12-LO mRNA expression in MC exposed to HG (12.71 ± 1.17 attm/μL) and DM G (1.78 ± 0.65 × 10-3 attm/glomerulus) was significantly higher than those of MCs in NG media (6.71 ± 0.78 attm/μL) and control G (0.34 ± 0.12 × 10-3 attm/glomerulus, P < 0.005), respectively. Western blot revealed a 1.7- and a 2.8-fold increase in MC and G 12-LO protein expression, respectively (P < 0.05). The immunohistochemistry score for G 12-LO and diabetic nephropathy score was significantly greater in DM and DM + I than controls. MC and G GAPDH mRNA remained unchanged. Conclusions. In MCs exposed to HG and in diabetic rat glomeruli, increments in 12-LO mRNA and protein are associated with changes modeling diabetic nephropathy. These findings suggest a role for the 12-LO pathway in the pathogenesis of diabetic nephropathy.

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