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A novel murine model to deplete hepatic stellate cells uncovers their role in amplifying liver damage in mice
Author(s) -
Puche Juan E.,
Lee Youngmin A.,
Jiao Jingjing,
Aloman Costica,
Fiel Maria I.,
Muñoz Ursula,
Kraus Thomas,
Lee Tingfang,
Yee Hal F.,
Friedman Scott L.
Publication year - 2013
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.26053
Subject(s) - hepatic stellate cell , sirius red , liver injury , hepatocyte , genetically modified mouse , fibrosis , hepatic fibrosis , microbiology and biotechnology , biology , chemistry , pathology , endocrinology , medicine , transgene , biochemistry , gene , in vitro
We have developed a novel model for depleting mouse hepatic stellate cells (HSCs) that has allowed us to clarify their contributions to hepatic injury and fibrosis. Transgenic (Tg) mice expressing the herpes simplex virus thymidine kinase gene ( HSV‐Tk ) driven by the mouse GFAP promoter were used to render proliferating HSCs susceptible to killing in response to ganciclovir (GCV). Effects of GCV were explored in primary HSCs and in vivo . Panlobular damage was provoked to maximize HSC depletion by combining CCl 4 (centrilobular injury) with allyl alcohol (AA) (periportal injury), as well as in a bile duct ligation (BDL) model. Cell depletion in situ was quantified using dual immunofluorescence (IF) for desmin and GFAP. In primary HSCs isolated from both untreated wild‐type (WT) and Tg mice, GCV induced cell death in ∼50% of HSCs from Tg, but not WT, mice. In TG mice treated with CCl 4 +AA+GCV, there was a significant decrease in GFAP and desmin‐positive cells, compared to WT mice (∼65% reduction; P < 0.01), which was accompanied by a decrease in the expression of HSC‐activation markers (alpha smooth muscle actin, beta platelet‐derived growth factor receptor, and collagen I). Similar results were observed after BDL. Associated with HSC depletion in both fibrosis models, there was marked attenuation of fibrosis and liver injury, as indicated by Sirius Red/Fast Green, hematoxylin and eosin quantification, and serum alanine/aspartate aminotransferase. Hepatic expression of interleukin‐10 and interferon‐gamma was increased after HSC depletion. No toxicity of GCV in either WT or Tg mice accounted for the differences in injury. Conclusion : Activated HSCs significantly amplify the response to liver injury, further expanding this cell type's repertoire in orchestrating hepatic injury and repair. (H EPATOLOGY 2013)