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

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

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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.

Original languageEnglish (US)
Pages (from-to)185-196
Number of pages12
JournalHepatology
Volume63
Issue number1
DOIs
StatePublished - Jan 1 2016
Externally publishedYes

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

ASJC Scopus subject areas

  • Hepatology

Cite this

Tabibian, J. H., O'Hara, S. P., Trussoni, C. E., Tietz, P. S., Splinter, P. L., Mounajjed, T., ... Larusso, N. F. (2016). Absence of the intestinal microbiota exacerbates hepatobiliary disease in a murine model of primary sclerosing cholangitis. Hepatology, 63(1), 185-196. https://doi.org/10.1002/hep.27927

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

In: Hepatology, Vol. 63, No. 1, 01.01.2016, p. 185-196.

Research output: Contribution to journalArticle

Tabibian, JH, O'Hara, SP, Trussoni, CE, Tietz, PS, Splinter, PL, Mounajjed, T, Hagey, LR & Larusso, NF 2016, 'Absence of the intestinal microbiota exacerbates hepatobiliary disease in a murine model of primary sclerosing cholangitis', Hepatology, vol. 63, no. 1, pp. 185-196. https://doi.org/10.1002/hep.27927
Tabibian, James H. ; O'Hara, Steven P. ; Trussoni, Christy E. ; Tietz, Pamela S. ; Splinter, Patrick L. ; Mounajjed, Taofic ; Hagey, Lee R. ; Larusso, Nicholas F. / Absence of the intestinal microbiota exacerbates hepatobiliary disease in a murine model of primary sclerosing cholangitis. In: Hepatology. 2016 ; Vol. 63, No. 1. pp. 185-196.
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AU - Tietz, Pamela S.

AU - Splinter, Patrick L.

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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.

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