Activated hepatic stellate cells and portal fibroblasts contribute to cholestatic liver fibrosis in MDR2 knockout mice

Takahiro Nishio, Ronglin Hu, Yukinori Koyama, Shuang Liang, Sara B. Rosenthal, Gen Yamamoto, Daniel Karin, Jacopo Baglieri, Hsiao Yen Ma, Jun Xu, Xiao Liu, Debanjan Dhar, K. Iwaisako, Kojiro Taura, David A. Brenner, Tatiana Kisseleva

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Background & Aims: Chronic liver injury often results in the activation of hepatic myofibroblasts and the development of liver fibrosis. Hepatic myofibroblasts may originate from 3 major sources: hepatic stellate cells (HSCs), portal fibroblasts (PFs), and fibrocytes, with varying contributions depending on the etiology of liver injury. Here, we assessed the composition of hepatic myofibroblasts in multidrug resistance gene 2 knockout (Mdr2−/−) mice, a genetic model that resembles primary sclerosing cholangitis in patients. Methods: Mdr2−/− mice expressing a collagen-GFP reporter were analyzed at different ages. Hepatic non-parenchymal cells isolated from collagen-GFP Mdr2−/− mice were sorted based on collagen-GFP and vitamin A. An NADPH oxidase (NOX) 1/4 inhibitor was administrated to Mdr2−/− mice aged 12–16 weeks old to assess the therapeutic approach of targeting oxidative stress in cholestatic injury. Results: Thy1+ activated PFs accounted for 26%, 51%, and 54% of collagen-GFP+ myofibroblasts in Mdr2−/− mice at 4, 8, and 16 weeks of age, respectively. The remaining collagen-GFP+ myofibroblasts were composed of activated HSCs, suggesting that PFs and HSCs are both activated in Mdr2−/− mice. Bone-marrow-derived fibrocytes minimally contributed to liver fibrosis in Mdr2−/− mice. The development of cholestatic liver fibrosis in Mdr2−/− mice was associated with early recruitment of Gr1+ myeloid cells and upregulation of pro-inflammatory cytokines (4 weeks). Administration of a NOX inhibitor to 12-week-old Mdr2−/− mice suppressed the activation of myofibroblasts and attenuated the development of cholestatic fibrosis. Conclusions: Activated PFs and activated HSCs contribute to cholestatic fibrosis in Mdr2−/− mice, and serve as targets for antifibrotic therapy. Lay summary: Activated portal fibroblasts and hepatic stellate cells, but not fibrocytes, contributed to the production of the fibrous scar in livers of Mdr2−/− mice, and these cells can serve as targets for antifibrotic therapy in cholestatic injury. Therapeutic inhibition of the enzyme NADPH oxidase (NOX) in Mdr2−/− mice reversed cholestatic fibrosis, suggesting that targeting NOXs may be an effective strategy for the treatment of cholestatic fibrosis.

Original languageEnglish (US)
Pages (from-to)573-585
Number of pages13
JournalJournal of Hepatology
Volume71
Issue number3
DOIs
StatePublished - Sep 2019
Externally publishedYes

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Hepatic Stellate Cells
Knockout Mice
Liver Cirrhosis
Fibroblasts
Myofibroblasts
Liver
Collagen
Fibrosis
NADPH Oxidase
Wounds and Injuries
MDR Genes
Sclerosing Cholangitis
Gene Knockout Techniques
Genetic Models
Myeloid Cells
Therapeutics
Vitamin A
Cicatrix
Oxidative Stress
Up-Regulation

Keywords

  • Activated hepatic stellate cells
  • Activated portal fibroblasts
  • Cholestatic fibrosis

ASJC Scopus subject areas

  • Hepatology

Cite this

Activated hepatic stellate cells and portal fibroblasts contribute to cholestatic liver fibrosis in MDR2 knockout mice. / Nishio, Takahiro; Hu, Ronglin; Koyama, Yukinori; Liang, Shuang; Rosenthal, Sara B.; Yamamoto, Gen; Karin, Daniel; Baglieri, Jacopo; Ma, Hsiao Yen; Xu, Jun; Liu, Xiao; Dhar, Debanjan; Iwaisako, K.; Taura, Kojiro; Brenner, David A.; Kisseleva, Tatiana.

In: Journal of Hepatology, Vol. 71, No. 3, 09.2019, p. 573-585.

Research output: Contribution to journalArticle

Nishio, T, Hu, R, Koyama, Y, Liang, S, Rosenthal, SB, Yamamoto, G, Karin, D, Baglieri, J, Ma, HY, Xu, J, Liu, X, Dhar, D, Iwaisako, K, Taura, K, Brenner, DA & Kisseleva, T 2019, 'Activated hepatic stellate cells and portal fibroblasts contribute to cholestatic liver fibrosis in MDR2 knockout mice', Journal of Hepatology, vol. 71, no. 3, pp. 573-585. https://doi.org/10.1016/j.jhep.2019.04.012
Nishio, Takahiro ; Hu, Ronglin ; Koyama, Yukinori ; Liang, Shuang ; Rosenthal, Sara B. ; Yamamoto, Gen ; Karin, Daniel ; Baglieri, Jacopo ; Ma, Hsiao Yen ; Xu, Jun ; Liu, Xiao ; Dhar, Debanjan ; Iwaisako, K. ; Taura, Kojiro ; Brenner, David A. ; Kisseleva, Tatiana. / Activated hepatic stellate cells and portal fibroblasts contribute to cholestatic liver fibrosis in MDR2 knockout mice. In: Journal of Hepatology. 2019 ; Vol. 71, No. 3. pp. 573-585.
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abstract = "Background & Aims: Chronic liver injury often results in the activation of hepatic myofibroblasts and the development of liver fibrosis. Hepatic myofibroblasts may originate from 3 major sources: hepatic stellate cells (HSCs), portal fibroblasts (PFs), and fibrocytes, with varying contributions depending on the etiology of liver injury. Here, we assessed the composition of hepatic myofibroblasts in multidrug resistance gene 2 knockout (Mdr2−/−) mice, a genetic model that resembles primary sclerosing cholangitis in patients. Methods: Mdr2−/− mice expressing a collagen-GFP reporter were analyzed at different ages. Hepatic non-parenchymal cells isolated from collagen-GFP Mdr2−/− mice were sorted based on collagen-GFP and vitamin A. An NADPH oxidase (NOX) 1/4 inhibitor was administrated to Mdr2−/− mice aged 12–16 weeks old to assess the therapeutic approach of targeting oxidative stress in cholestatic injury. Results: Thy1+ activated PFs accounted for 26{\%}, 51{\%}, and 54{\%} of collagen-GFP+ myofibroblasts in Mdr2−/− mice at 4, 8, and 16 weeks of age, respectively. The remaining collagen-GFP+ myofibroblasts were composed of activated HSCs, suggesting that PFs and HSCs are both activated in Mdr2−/− mice. Bone-marrow-derived fibrocytes minimally contributed to liver fibrosis in Mdr2−/− mice. The development of cholestatic liver fibrosis in Mdr2−/− mice was associated with early recruitment of Gr1+ myeloid cells and upregulation of pro-inflammatory cytokines (4 weeks). Administration of a NOX inhibitor to 12-week-old Mdr2−/− mice suppressed the activation of myofibroblasts and attenuated the development of cholestatic fibrosis. Conclusions: Activated PFs and activated HSCs contribute to cholestatic fibrosis in Mdr2−/− mice, and serve as targets for antifibrotic therapy. Lay summary: Activated portal fibroblasts and hepatic stellate cells, but not fibrocytes, contributed to the production of the fibrous scar in livers of Mdr2−/− mice, and these cells can serve as targets for antifibrotic therapy in cholestatic injury. Therapeutic inhibition of the enzyme NADPH oxidase (NOX) in Mdr2−/− mice reversed cholestatic fibrosis, suggesting that targeting NOXs may be an effective strategy for the treatment of cholestatic fibrosis.",
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author = "Takahiro Nishio and Ronglin Hu and Yukinori Koyama and Shuang Liang and Rosenthal, {Sara B.} and Gen Yamamoto and Daniel Karin and Jacopo Baglieri and Ma, {Hsiao Yen} and Jun Xu and Xiao Liu and Debanjan Dhar and K. Iwaisako and Kojiro Taura and Brenner, {David A.} and Tatiana Kisseleva",
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TY - JOUR

T1 - Activated hepatic stellate cells and portal fibroblasts contribute to cholestatic liver fibrosis in MDR2 knockout mice

AU - Nishio, Takahiro

AU - Hu, Ronglin

AU - Koyama, Yukinori

AU - Liang, Shuang

AU - Rosenthal, Sara B.

AU - Yamamoto, Gen

AU - Karin, Daniel

AU - Baglieri, Jacopo

AU - Ma, Hsiao Yen

AU - Xu, Jun

AU - Liu, Xiao

AU - Dhar, Debanjan

AU - Iwaisako, K.

AU - Taura, Kojiro

AU - Brenner, David A.

AU - Kisseleva, Tatiana

PY - 2019/9

Y1 - 2019/9

N2 - Background & Aims: Chronic liver injury often results in the activation of hepatic myofibroblasts and the development of liver fibrosis. Hepatic myofibroblasts may originate from 3 major sources: hepatic stellate cells (HSCs), portal fibroblasts (PFs), and fibrocytes, with varying contributions depending on the etiology of liver injury. Here, we assessed the composition of hepatic myofibroblasts in multidrug resistance gene 2 knockout (Mdr2−/−) mice, a genetic model that resembles primary sclerosing cholangitis in patients. Methods: Mdr2−/− mice expressing a collagen-GFP reporter were analyzed at different ages. Hepatic non-parenchymal cells isolated from collagen-GFP Mdr2−/− mice were sorted based on collagen-GFP and vitamin A. An NADPH oxidase (NOX) 1/4 inhibitor was administrated to Mdr2−/− mice aged 12–16 weeks old to assess the therapeutic approach of targeting oxidative stress in cholestatic injury. Results: Thy1+ activated PFs accounted for 26%, 51%, and 54% of collagen-GFP+ myofibroblasts in Mdr2−/− mice at 4, 8, and 16 weeks of age, respectively. The remaining collagen-GFP+ myofibroblasts were composed of activated HSCs, suggesting that PFs and HSCs are both activated in Mdr2−/− mice. Bone-marrow-derived fibrocytes minimally contributed to liver fibrosis in Mdr2−/− mice. The development of cholestatic liver fibrosis in Mdr2−/− mice was associated with early recruitment of Gr1+ myeloid cells and upregulation of pro-inflammatory cytokines (4 weeks). Administration of a NOX inhibitor to 12-week-old Mdr2−/− mice suppressed the activation of myofibroblasts and attenuated the development of cholestatic fibrosis. Conclusions: Activated PFs and activated HSCs contribute to cholestatic fibrosis in Mdr2−/− mice, and serve as targets for antifibrotic therapy. Lay summary: Activated portal fibroblasts and hepatic stellate cells, but not fibrocytes, contributed to the production of the fibrous scar in livers of Mdr2−/− mice, and these cells can serve as targets for antifibrotic therapy in cholestatic injury. Therapeutic inhibition of the enzyme NADPH oxidase (NOX) in Mdr2−/− mice reversed cholestatic fibrosis, suggesting that targeting NOXs may be an effective strategy for the treatment of cholestatic fibrosis.

AB - Background & Aims: Chronic liver injury often results in the activation of hepatic myofibroblasts and the development of liver fibrosis. Hepatic myofibroblasts may originate from 3 major sources: hepatic stellate cells (HSCs), portal fibroblasts (PFs), and fibrocytes, with varying contributions depending on the etiology of liver injury. Here, we assessed the composition of hepatic myofibroblasts in multidrug resistance gene 2 knockout (Mdr2−/−) mice, a genetic model that resembles primary sclerosing cholangitis in patients. Methods: Mdr2−/− mice expressing a collagen-GFP reporter were analyzed at different ages. Hepatic non-parenchymal cells isolated from collagen-GFP Mdr2−/− mice were sorted based on collagen-GFP and vitamin A. An NADPH oxidase (NOX) 1/4 inhibitor was administrated to Mdr2−/− mice aged 12–16 weeks old to assess the therapeutic approach of targeting oxidative stress in cholestatic injury. Results: Thy1+ activated PFs accounted for 26%, 51%, and 54% of collagen-GFP+ myofibroblasts in Mdr2−/− mice at 4, 8, and 16 weeks of age, respectively. The remaining collagen-GFP+ myofibroblasts were composed of activated HSCs, suggesting that PFs and HSCs are both activated in Mdr2−/− mice. Bone-marrow-derived fibrocytes minimally contributed to liver fibrosis in Mdr2−/− mice. The development of cholestatic liver fibrosis in Mdr2−/− mice was associated with early recruitment of Gr1+ myeloid cells and upregulation of pro-inflammatory cytokines (4 weeks). Administration of a NOX inhibitor to 12-week-old Mdr2−/− mice suppressed the activation of myofibroblasts and attenuated the development of cholestatic fibrosis. Conclusions: Activated PFs and activated HSCs contribute to cholestatic fibrosis in Mdr2−/− mice, and serve as targets for antifibrotic therapy. Lay summary: Activated portal fibroblasts and hepatic stellate cells, but not fibrocytes, contributed to the production of the fibrous scar in livers of Mdr2−/− mice, and these cells can serve as targets for antifibrotic therapy in cholestatic injury. Therapeutic inhibition of the enzyme NADPH oxidase (NOX) in Mdr2−/− mice reversed cholestatic fibrosis, suggesting that targeting NOXs may be an effective strategy for the treatment of cholestatic fibrosis.

KW - Activated hepatic stellate cells

KW - Activated portal fibroblasts

KW - Cholestatic fibrosis

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