Xanthine oxidase-derived free radicals directly activate rat pancreatic stellate cells

Hiroaki Tanioka, Takaaki Mizushima, Akinori Shirahige, Koki Matsushita, Koji Ochi, Mitsuko Ichimura, Naoki Matsumura, Toshiyuki Shinji, Mitsune Tanimoto, Norio Koide

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Background and Aim: Free radicals are reported to be associated with fibrosis in the pancreas. It is generally accepted that pancreatic stellate cells (PSC) play an important role in pancreatic fibrosis. However, the exact role of free radicals in activation of PSC has not been fully elucidated. In the present study, using a superoxide dismutase (SOD) inhibitor, diethyldithiocarbamate (DDC) with cultured PSC, we investigated how free radicals act on the activation of PSC. Methods: PSC were isolated from male Wister rats. Cultured rat PSC were incubated with DDC for 48 h. Intracellular SOD activity and lipid peroxidation were examined in DDC-treated PSC. Activation of PSC was examined by determining the expression of α-smooth muscle actin (α-SMA) by immunocytochemistry. The number of PSC using a hemocytometer, type I collagen secretion with ELISA and matrix metalloproteinases (MMP) activities with gelatin zymography were also examined. Secretion of transforming growth factor-β1 (TGF-β1) was evaluated by ELISA. The effects of the allopurinol, a xanthine oxidase (XOD) inhibitor, on PSC were also examined. Results: DDC decreased SOD activity and increased lipid peroxidation products in PSC. DDC activated PSC, increasing the number of α-SMA positive cells, enhancing secretion of type I collagen and MMP, inhibiting PSC proliferation. Secretion of TGF-β1, which is known to activate PSC, was increased by DDC treatment. These alterations were prevented by allopurinol. Conclusion: These results suggest that free radicals generated by XOD might directly activate PSC.

Original languageEnglish
Pages (from-to)537-544
Number of pages8
JournalJournal of Gastroenterology and Hepatology (Australia)
Volume21
Issue number3
DOIs
Publication statusPublished - 2006

Fingerprint

Pancreatic Stellate Cells
Xanthine Oxidase
Free Radicals
Ditiocarb
Superoxide Dismutase
Allopurinol
Transforming Growth Factors
Collagen Type I
Matrix Metalloproteinases
Lipid Peroxidation
Fibrosis
Enzyme-Linked Immunosorbent Assay

Keywords

  • Diethyldithiocarbamate
  • Fibrosis
  • Free radical
  • Pancreatic stellate cell
  • Superoxide dismutase

ASJC Scopus subject areas

  • Hepatology
  • Gastroenterology

Cite this

Xanthine oxidase-derived free radicals directly activate rat pancreatic stellate cells. / Tanioka, Hiroaki; Mizushima, Takaaki; Shirahige, Akinori; Matsushita, Koki; Ochi, Koji; Ichimura, Mitsuko; Matsumura, Naoki; Shinji, Toshiyuki; Tanimoto, Mitsune; Koide, Norio.

In: Journal of Gastroenterology and Hepatology (Australia), Vol. 21, No. 3, 2006, p. 537-544.

Research output: Contribution to journalArticle

Tanioka, H, Mizushima, T, Shirahige, A, Matsushita, K, Ochi, K, Ichimura, M, Matsumura, N, Shinji, T, Tanimoto, M & Koide, N 2006, 'Xanthine oxidase-derived free radicals directly activate rat pancreatic stellate cells', Journal of Gastroenterology and Hepatology (Australia), vol. 21, no. 3, pp. 537-544. https://doi.org/10.1111/j.1440-1746.2005.03999.x
Tanioka, Hiroaki ; Mizushima, Takaaki ; Shirahige, Akinori ; Matsushita, Koki ; Ochi, Koji ; Ichimura, Mitsuko ; Matsumura, Naoki ; Shinji, Toshiyuki ; Tanimoto, Mitsune ; Koide, Norio. / Xanthine oxidase-derived free radicals directly activate rat pancreatic stellate cells. In: Journal of Gastroenterology and Hepatology (Australia). 2006 ; Vol. 21, No. 3. pp. 537-544.
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abstract = "Background and Aim: Free radicals are reported to be associated with fibrosis in the pancreas. It is generally accepted that pancreatic stellate cells (PSC) play an important role in pancreatic fibrosis. However, the exact role of free radicals in activation of PSC has not been fully elucidated. In the present study, using a superoxide dismutase (SOD) inhibitor, diethyldithiocarbamate (DDC) with cultured PSC, we investigated how free radicals act on the activation of PSC. Methods: PSC were isolated from male Wister rats. Cultured rat PSC were incubated with DDC for 48 h. Intracellular SOD activity and lipid peroxidation were examined in DDC-treated PSC. Activation of PSC was examined by determining the expression of α-smooth muscle actin (α-SMA) by immunocytochemistry. The number of PSC using a hemocytometer, type I collagen secretion with ELISA and matrix metalloproteinases (MMP) activities with gelatin zymography were also examined. Secretion of transforming growth factor-β1 (TGF-β1) was evaluated by ELISA. The effects of the allopurinol, a xanthine oxidase (XOD) inhibitor, on PSC were also examined. Results: DDC decreased SOD activity and increased lipid peroxidation products in PSC. DDC activated PSC, increasing the number of α-SMA positive cells, enhancing secretion of type I collagen and MMP, inhibiting PSC proliferation. Secretion of TGF-β1, which is known to activate PSC, was increased by DDC treatment. These alterations were prevented by allopurinol. Conclusion: These results suggest that free radicals generated by XOD might directly activate PSC.",
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AU - Tanioka, Hiroaki

AU - Mizushima, Takaaki

AU - Shirahige, Akinori

AU - Matsushita, Koki

AU - Ochi, Koji

AU - Ichimura, Mitsuko

AU - Matsumura, Naoki

AU - Shinji, Toshiyuki

AU - Tanimoto, Mitsune

AU - Koide, Norio

PY - 2006

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N2 - Background and Aim: Free radicals are reported to be associated with fibrosis in the pancreas. It is generally accepted that pancreatic stellate cells (PSC) play an important role in pancreatic fibrosis. However, the exact role of free radicals in activation of PSC has not been fully elucidated. In the present study, using a superoxide dismutase (SOD) inhibitor, diethyldithiocarbamate (DDC) with cultured PSC, we investigated how free radicals act on the activation of PSC. Methods: PSC were isolated from male Wister rats. Cultured rat PSC were incubated with DDC for 48 h. Intracellular SOD activity and lipid peroxidation were examined in DDC-treated PSC. Activation of PSC was examined by determining the expression of α-smooth muscle actin (α-SMA) by immunocytochemistry. The number of PSC using a hemocytometer, type I collagen secretion with ELISA and matrix metalloproteinases (MMP) activities with gelatin zymography were also examined. Secretion of transforming growth factor-β1 (TGF-β1) was evaluated by ELISA. The effects of the allopurinol, a xanthine oxidase (XOD) inhibitor, on PSC were also examined. Results: DDC decreased SOD activity and increased lipid peroxidation products in PSC. DDC activated PSC, increasing the number of α-SMA positive cells, enhancing secretion of type I collagen and MMP, inhibiting PSC proliferation. Secretion of TGF-β1, which is known to activate PSC, was increased by DDC treatment. These alterations were prevented by allopurinol. Conclusion: These results suggest that free radicals generated by XOD might directly activate PSC.

AB - Background and Aim: Free radicals are reported to be associated with fibrosis in the pancreas. It is generally accepted that pancreatic stellate cells (PSC) play an important role in pancreatic fibrosis. However, the exact role of free radicals in activation of PSC has not been fully elucidated. In the present study, using a superoxide dismutase (SOD) inhibitor, diethyldithiocarbamate (DDC) with cultured PSC, we investigated how free radicals act on the activation of PSC. Methods: PSC were isolated from male Wister rats. Cultured rat PSC were incubated with DDC for 48 h. Intracellular SOD activity and lipid peroxidation were examined in DDC-treated PSC. Activation of PSC was examined by determining the expression of α-smooth muscle actin (α-SMA) by immunocytochemistry. The number of PSC using a hemocytometer, type I collagen secretion with ELISA and matrix metalloproteinases (MMP) activities with gelatin zymography were also examined. Secretion of transforming growth factor-β1 (TGF-β1) was evaluated by ELISA. The effects of the allopurinol, a xanthine oxidase (XOD) inhibitor, on PSC were also examined. Results: DDC decreased SOD activity and increased lipid peroxidation products in PSC. DDC activated PSC, increasing the number of α-SMA positive cells, enhancing secretion of type I collagen and MMP, inhibiting PSC proliferation. Secretion of TGF-β1, which is known to activate PSC, was increased by DDC treatment. These alterations were prevented by allopurinol. Conclusion: These results suggest that free radicals generated by XOD might directly activate PSC.

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KW - Free radical

KW - Pancreatic stellate cell

KW - Superoxide dismutase

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