Absence of the intestinal microbiota exacerbates hepatobiliary disease in a murine model of primary sclerosing cholangitis in mice

James H. Tabibian, Steven P. O'Hara, Christy E. Trussoni, Pamela S. Tietz, Patrick L. Splinter, Taofic Mounajjed, Lee R. Hagey, Nicholas F La Russo

Research output: Contribution to journalArticle

81 Citations (Scopus)

Abstract

Primary sclerosing cholangitis (PSC) is a chronic, idiopathic, fibroinflammatory cholangiopathy. The role of the microbiota in PSC etiopathogenesis may be fundamentally important, yet remains obscure. We tested the hypothesis that germ-free (GF) mutltidrug resistance 2 knockout (mdr2<sup>-/-</sup>) mice develop a distinct PSC phenotype, compared to conventionally housed (CV) mdr2<sup>-/-</sup> mice. Mdr2<sup>-/-</sup> mice (n=12) were rederived as GF by embryo transfer, maintained in isolators, and sacrificed at 60 days in parallel with age-matched CV mdr2<sup>-/-</sup> mice. Serum biochemistries, gallbladder bile acids, and liver sections were examined. Histological findings were validated morphometrically, biochemically, and by immunofluorescence microscopy (IFM). Cholangiocyte senescence was assessed by p16<sup>INK4a</sup> in situ hybridization in liver tissue and by senescence-associated β-galactosidase staining in a culture-based model of insult-induced senescence. Serum biochemistries, including alkaline phosphatase, aspartate aminotransferase, and bilirubin, were significantly higher in GF mdr2<sup>-/-</sup> (P<0.01). Primary bile acids were similar, whereas secondary bile acids were absent, in GF mdr2<sup>-/-</sup> mice. Fibrosis, ductular reaction, and ductopenia were significantly more severe histopathologically in GF mdr2<sup>-/-</sup> mice (P<0.01) and were confirmed by hepatic morphometry, hydroxyproline assay, and IFM. Cholangiocyte senescence was significantly increased in GF mdr2<sup>-/-</sup> mice and abrogated in vitro by ursodeoxycholic acid (UDCA) treatment. Conclusions: GF mdr2<sup>-/-</sup> mice exhibit exacerbated biochemical and histological features of PSC and increased cholangiocyte senescence, a characteristic and potential mediator of progressive biliary disease. UDCA, a commensal microbial metabolite, abrogates senescence in vitro. These findings demonstrate the importance of the commensal microbiota and its metabolites in protecting against biliary injury and suggest avenues for future studies of biomarkers and therapeutic interventions in PSC.

Original languageEnglish (US)
JournalHepatology
DOIs
StateAccepted/In press - 2015

Fingerprint

Sclerosing Cholangitis
Ursodeoxycholic Acid
Microbiota
Biochemistry
Galactosidases
Liver
Embryo Transfer
Aspartate Aminotransferases
Gastrointestinal Microbiome
Gallbladder
Bile Acids and Salts
Serum
Fluorescence Microscopy
Bilirubin
Knockout Mice
In Situ Hybridization
Alkaline Phosphatase
Fibrosis
Biomarkers
Staining and Labeling

ASJC Scopus subject areas

  • Hepatology

Cite this

Absence of the intestinal microbiota exacerbates hepatobiliary disease in a murine model of primary sclerosing cholangitis in mice. / Tabibian, James H.; O'Hara, Steven P.; Trussoni, Christy E.; Tietz, Pamela S.; Splinter, Patrick L.; Mounajjed, Taofic; Hagey, Lee R.; La Russo, Nicholas F.

In: Hepatology, 2015.

Research output: Contribution to journalArticle

Tabibian, James H. ; O'Hara, Steven P. ; Trussoni, Christy E. ; Tietz, Pamela S. ; Splinter, Patrick L. ; Mounajjed, Taofic ; Hagey, Lee R. ; La Russo, Nicholas F. / Absence of the intestinal microbiota exacerbates hepatobiliary disease in a murine model of primary sclerosing cholangitis in mice. In: Hepatology. 2015.
@article{b4eae075ff2d41359f9f976136eed77c,
title = "Absence of the intestinal microbiota exacerbates hepatobiliary disease in a murine model of primary sclerosing cholangitis in mice",
abstract = "Primary sclerosing cholangitis (PSC) is a chronic, idiopathic, fibroinflammatory cholangiopathy. The role of the microbiota in PSC etiopathogenesis may be fundamentally important, yet remains obscure. We tested the hypothesis that germ-free (GF) mutltidrug resistance 2 knockout (mdr2-/-) mice develop a distinct PSC phenotype, compared to conventionally housed (CV) mdr2-/- mice. Mdr2-/- mice (n=12) were rederived as GF by embryo transfer, maintained in isolators, and sacrificed at 60 days in parallel with age-matched CV mdr2-/- mice. Serum biochemistries, gallbladder bile acids, and liver sections were examined. Histological findings were validated morphometrically, biochemically, and by immunofluorescence microscopy (IFM). Cholangiocyte senescence was assessed by p16INK4a in situ hybridization in liver tissue and by senescence-associated β-galactosidase staining in a culture-based model of insult-induced senescence. Serum biochemistries, including alkaline phosphatase, aspartate aminotransferase, and bilirubin, were significantly higher in GF mdr2-/- (P<0.01). Primary bile acids were similar, whereas secondary bile acids were absent, in GF mdr2-/- mice. Fibrosis, ductular reaction, and ductopenia were significantly more severe histopathologically in GF mdr2-/- mice (P<0.01) and were confirmed by hepatic morphometry, hydroxyproline assay, and IFM. Cholangiocyte senescence was significantly increased in GF mdr2-/- mice and abrogated in vitro by ursodeoxycholic acid (UDCA) treatment. Conclusions: GF mdr2-/- mice exhibit exacerbated biochemical and histological features of PSC and increased cholangiocyte senescence, a characteristic and potential mediator of progressive biliary disease. UDCA, a commensal microbial metabolite, abrogates senescence in vitro. These findings demonstrate the importance of the commensal microbiota and its metabolites in protecting against biliary injury and suggest avenues for future studies of biomarkers and therapeutic interventions in PSC.",
author = "Tabibian, {James H.} and O'Hara, {Steven P.} and Trussoni, {Christy E.} and Tietz, {Pamela S.} and Splinter, {Patrick L.} and Taofic Mounajjed and Hagey, {Lee R.} and {La Russo}, {Nicholas F}",
year = "2015",
doi = "10.1002/hep.27927",
language = "English (US)",
journal = "Hepatology",
issn = "0270-9139",
publisher = "John Wiley and Sons Ltd",

}

TY - JOUR

T1 - Absence of the intestinal microbiota exacerbates hepatobiliary disease in a murine model of primary sclerosing cholangitis in mice

AU - Tabibian, James H.

AU - O'Hara, Steven P.

AU - Trussoni, Christy E.

AU - Tietz, Pamela S.

AU - Splinter, Patrick L.

AU - Mounajjed, Taofic

AU - Hagey, Lee R.

AU - La Russo, Nicholas F

PY - 2015

Y1 - 2015

N2 - Primary sclerosing cholangitis (PSC) is a chronic, idiopathic, fibroinflammatory cholangiopathy. The role of the microbiota in PSC etiopathogenesis may be fundamentally important, yet remains obscure. We tested the hypothesis that germ-free (GF) mutltidrug resistance 2 knockout (mdr2-/-) mice develop a distinct PSC phenotype, compared to conventionally housed (CV) mdr2-/- mice. Mdr2-/- mice (n=12) were rederived as GF by embryo transfer, maintained in isolators, and sacrificed at 60 days in parallel with age-matched CV mdr2-/- mice. Serum biochemistries, gallbladder bile acids, and liver sections were examined. Histological findings were validated morphometrically, biochemically, and by immunofluorescence microscopy (IFM). Cholangiocyte senescence was assessed by p16INK4a in situ hybridization in liver tissue and by senescence-associated β-galactosidase staining in a culture-based model of insult-induced senescence. Serum biochemistries, including alkaline phosphatase, aspartate aminotransferase, and bilirubin, were significantly higher in GF mdr2-/- (P<0.01). Primary bile acids were similar, whereas secondary bile acids were absent, in GF mdr2-/- mice. Fibrosis, ductular reaction, and ductopenia were significantly more severe histopathologically in GF mdr2-/- mice (P<0.01) and were confirmed by hepatic morphometry, hydroxyproline assay, and IFM. Cholangiocyte senescence was significantly increased in GF mdr2-/- mice and abrogated in vitro by ursodeoxycholic acid (UDCA) treatment. Conclusions: GF mdr2-/- mice exhibit exacerbated biochemical and histological features of PSC and increased cholangiocyte senescence, a characteristic and potential mediator of progressive biliary disease. UDCA, a commensal microbial metabolite, abrogates senescence in vitro. These findings demonstrate the importance of the commensal microbiota and its metabolites in protecting against biliary injury and suggest avenues for future studies of biomarkers and therapeutic interventions in PSC.

AB - Primary sclerosing cholangitis (PSC) is a chronic, idiopathic, fibroinflammatory cholangiopathy. The role of the microbiota in PSC etiopathogenesis may be fundamentally important, yet remains obscure. We tested the hypothesis that germ-free (GF) mutltidrug resistance 2 knockout (mdr2-/-) mice develop a distinct PSC phenotype, compared to conventionally housed (CV) mdr2-/- mice. Mdr2-/- mice (n=12) were rederived as GF by embryo transfer, maintained in isolators, and sacrificed at 60 days in parallel with age-matched CV mdr2-/- mice. Serum biochemistries, gallbladder bile acids, and liver sections were examined. Histological findings were validated morphometrically, biochemically, and by immunofluorescence microscopy (IFM). Cholangiocyte senescence was assessed by p16INK4a in situ hybridization in liver tissue and by senescence-associated β-galactosidase staining in a culture-based model of insult-induced senescence. Serum biochemistries, including alkaline phosphatase, aspartate aminotransferase, and bilirubin, were significantly higher in GF mdr2-/- (P<0.01). Primary bile acids were similar, whereas secondary bile acids were absent, in GF mdr2-/- mice. Fibrosis, ductular reaction, and ductopenia were significantly more severe histopathologically in GF mdr2-/- mice (P<0.01) and were confirmed by hepatic morphometry, hydroxyproline assay, and IFM. Cholangiocyte senescence was significantly increased in GF mdr2-/- mice and abrogated in vitro by ursodeoxycholic acid (UDCA) treatment. Conclusions: GF mdr2-/- mice exhibit exacerbated biochemical and histological features of PSC and increased cholangiocyte senescence, a characteristic and potential mediator of progressive biliary disease. UDCA, a commensal microbial metabolite, abrogates senescence in vitro. These findings demonstrate the importance of the commensal microbiota and its metabolites in protecting against biliary injury and suggest avenues for future studies of biomarkers and therapeutic interventions in PSC.

UR - http://www.scopus.com/inward/record.url?scp=84938890827&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84938890827&partnerID=8YFLogxK

U2 - 10.1002/hep.27927

DO - 10.1002/hep.27927

M3 - Article

C2 - 26044703

AN - SCOPUS:84952637577

JO - Hepatology

JF - Hepatology

SN - 0270-9139

ER -