Fasudil hydrochloride hydrate, a Rho-kinase (ROCK) inhibitor, suppresses collagen production and enhances collagenase activity in hepatic stellate cells

Marie Fukushima, Makoto Nakamuta, Motoyuki Kohjima, Kazuhiro Kotoh, Munechika Enjoji, Naoya Kobayashi, Hajime Nawata

Research output: Contribution to journalArticle

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Abstract

Background/Aims: The Rho-ROCK signaling pathways play an important role in the activation of hepatic stellate cells (HSCs). We investigated the effects of fasudil hydrochloride hydrate (fasudil), a Rho-kinase (ROCK) inhibitor, on cell growth, collagen production, and collagenase activity in HSCs. Methods: Rat HSCs and human HSC-derived TWNT-4 cells were cultured for studies on stress fiber formation and α-smooth muscle actin (α-SMA) expression. Proliferation was measured by BrdU incorporation, and apoptosis by TUNEL assay. The phosphorylation states of the MAP kinases (MAPKs), extra cellular signal-regulated kinase 1/2 (ERK1/2), c-jun kinase (JNK), and p38 were evaluated by western blot analysis. Type I collagen, matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) production and gene expression were evaluated by ELISA and real-time PCR, respectively. Collagenase activity (active MMP-1) was also evaluated. Results: Fasudil (100 μM) inhibited cell spreading, the formation of stress fibers, and expression of α-SMA with concomitant suppression of cell growth, although it did not induce apoptosis. Fasudil inhibited phosphorylation of ERK1/2, JNK, and p38. Treatment with fasudil suppressed the production and transcription of collagen and TIMP, stimulated the production and transcription of MMP-1, and enhanced collagenase activity. Conclusion: These findings demonstrated that fasudil not only suppresses proliferation and collagen production but also increases collagenase activity.

Original languageEnglish
Pages (from-to)829-838
Number of pages10
JournalLiver International
Volume25
Issue number4
DOIs
Publication statusPublished - Aug 1 2005

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Hepatic Stellate Cells
rho-Associated Kinases
Collagenases
Matrix Metalloproteinase 1
Collagen
Phosphotransferases
Stress Fibers
Mitogen-Activated Protein Kinase 9
Mitogen-Activated Protein Kinase 8
Phosphorylation
Apoptosis
Growth Inhibitors
Tissue Inhibitor of Metalloproteinase-1
Matrix Metalloproteinase Inhibitors
In Situ Nick-End Labeling
Bromodeoxyuridine
Collagen Type I
Smooth Muscle
fasudil
Actins

All Science Journal Classification (ASJC) codes

  • Hepatology

Cite this

Fasudil hydrochloride hydrate, a Rho-kinase (ROCK) inhibitor, suppresses collagen production and enhances collagenase activity in hepatic stellate cells. / Fukushima, Marie; Nakamuta, Makoto; Kohjima, Motoyuki; Kotoh, Kazuhiro; Enjoji, Munechika; Kobayashi, Naoya; Nawata, Hajime.

In: Liver International, Vol. 25, No. 4, 01.08.2005, p. 829-838.

Research output: Contribution to journalArticle

Fukushima, Marie ; Nakamuta, Makoto ; Kohjima, Motoyuki ; Kotoh, Kazuhiro ; Enjoji, Munechika ; Kobayashi, Naoya ; Nawata, Hajime. / Fasudil hydrochloride hydrate, a Rho-kinase (ROCK) inhibitor, suppresses collagen production and enhances collagenase activity in hepatic stellate cells. In: Liver International. 2005 ; Vol. 25, No. 4. pp. 829-838.
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AU - Nakamuta, Makoto

AU - Kohjima, Motoyuki

AU - Kotoh, Kazuhiro

AU - Enjoji, Munechika

AU - Kobayashi, Naoya

AU - Nawata, Hajime

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AB - Background/Aims: The Rho-ROCK signaling pathways play an important role in the activation of hepatic stellate cells (HSCs). We investigated the effects of fasudil hydrochloride hydrate (fasudil), a Rho-kinase (ROCK) inhibitor, on cell growth, collagen production, and collagenase activity in HSCs. Methods: Rat HSCs and human HSC-derived TWNT-4 cells were cultured for studies on stress fiber formation and α-smooth muscle actin (α-SMA) expression. Proliferation was measured by BrdU incorporation, and apoptosis by TUNEL assay. The phosphorylation states of the MAP kinases (MAPKs), extra cellular signal-regulated kinase 1/2 (ERK1/2), c-jun kinase (JNK), and p38 were evaluated by western blot analysis. Type I collagen, matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) production and gene expression were evaluated by ELISA and real-time PCR, respectively. Collagenase activity (active MMP-1) was also evaluated. Results: Fasudil (100 μM) inhibited cell spreading, the formation of stress fibers, and expression of α-SMA with concomitant suppression of cell growth, although it did not induce apoptosis. Fasudil inhibited phosphorylation of ERK1/2, JNK, and p38. Treatment with fasudil suppressed the production and transcription of collagen and TIMP, stimulated the production and transcription of MMP-1, and enhanced collagenase activity. Conclusion: These findings demonstrated that fasudil not only suppresses proliferation and collagen production but also increases collagenase activity.

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