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Hedgehog signalling regulates liver sinusoidal endothelial cell capillarisation.
Gut 2013; 62(2):299-309Gut

Abstract

OBJECTIVE

Vascular remodelling during liver damage involves loss of healthy liver sinusoidal endothelial cell (LSEC) phenotype via capillarisation. Hedgehog (Hh) signalling regulates vascular development and increases during liver injury. This study therefore examined its role in capillarisation.

DESIGN

Primary LSEC were cultured for 5 days to induce capillarisation. Pharmacological, antibody-mediated and genetic approaches were used to manipulate Hh signalling. Effects on mRNA and protein expression of Hh-regulated genes and capillarisation markers were evaluated by quantitative reverse transcription PCR and immunoblot. Changes in LSEC function were assessed by migration and tube forming assay, and gain/loss of fenestrae was examined by electron microscopy. Mice with acute or chronic liver injury were treated with Hh inhibitors; effects on capillarisation were assessed by immunohistochemistry.

RESULTS

Freshly isolated LSEC expressed Hh ligands, Hh receptors and Hh ligand antagonist Hhip. Capillarisation was accompanied by repression of Hhip and increased expression of Hh-regulated genes. Treatment with Hh agonist further induced expression of Hh ligands and Hh-regulated genes, and upregulated capillarisation-associated genes; whereas Hh signalling antagonist or Hh ligand neutralising antibody each repressed expression of Hh target genes and capillarisation markers. LSEC isolated from Smo(loxP/loxP) transgenic mice that had been infected with adenovirus expressing Cre-recombinase to delete Smoothened showed over 75% knockdown of Smoothened. During culture, Smoothened-deficient LSEC had inhibited Hh signalling, less induction of capillarisation-associated genes and retention of fenestrae. In mice with injured livers, inhibiting Hh signalling prevented capillarisation.

CONCLUSIONS

LSEC produce and respond to Hh ligands, and use Hh signalling to regulate complex phenotypic changes that occur during capillarisation.

Authors+Show Affiliations

Division of Gastroenterology Duke University Medical Center, Durham, NC 27710, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

22362915

Citation

Xie, Guanhua, et al. "Hedgehog Signalling Regulates Liver Sinusoidal Endothelial Cell Capillarisation." Gut, vol. 62, no. 2, 2013, pp. 299-309.
Xie G, Choi SS, Syn WK, et al. Hedgehog signalling regulates liver sinusoidal endothelial cell capillarisation. Gut. 2013;62(2):299-309.
Xie, G., Choi, S. S., Syn, W. K., Michelotti, G. A., Swiderska, M., Karaca, G., ... Diehl, A. M. (2013). Hedgehog signalling regulates liver sinusoidal endothelial cell capillarisation. Gut, 62(2), pp. 299-309. doi:10.1136/gutjnl-2011-301494.
Xie G, et al. Hedgehog Signalling Regulates Liver Sinusoidal Endothelial Cell Capillarisation. Gut. 2013;62(2):299-309. PubMed PMID: 22362915.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Hedgehog signalling regulates liver sinusoidal endothelial cell capillarisation. AU - Xie,Guanhua, AU - Choi,Steve S, AU - Syn,Wing-Kin, AU - Michelotti,Gregory A, AU - Swiderska,Marzena, AU - Karaca,Gamze, AU - Chan,Isaac S, AU - Chen,Yuping, AU - Diehl,Anna Mae, Y1 - 2012/02/23/ PY - 2012/2/25/entrez PY - 2012/3/1/pubmed PY - 2013/3/6/medline SP - 299 EP - 309 JF - Gut JO - Gut VL - 62 IS - 2 N2 - OBJECTIVE: Vascular remodelling during liver damage involves loss of healthy liver sinusoidal endothelial cell (LSEC) phenotype via capillarisation. Hedgehog (Hh) signalling regulates vascular development and increases during liver injury. This study therefore examined its role in capillarisation. DESIGN: Primary LSEC were cultured for 5 days to induce capillarisation. Pharmacological, antibody-mediated and genetic approaches were used to manipulate Hh signalling. Effects on mRNA and protein expression of Hh-regulated genes and capillarisation markers were evaluated by quantitative reverse transcription PCR and immunoblot. Changes in LSEC function were assessed by migration and tube forming assay, and gain/loss of fenestrae was examined by electron microscopy. Mice with acute or chronic liver injury were treated with Hh inhibitors; effects on capillarisation were assessed by immunohistochemistry. RESULTS: Freshly isolated LSEC expressed Hh ligands, Hh receptors and Hh ligand antagonist Hhip. Capillarisation was accompanied by repression of Hhip and increased expression of Hh-regulated genes. Treatment with Hh agonist further induced expression of Hh ligands and Hh-regulated genes, and upregulated capillarisation-associated genes; whereas Hh signalling antagonist or Hh ligand neutralising antibody each repressed expression of Hh target genes and capillarisation markers. LSEC isolated from Smo(loxP/loxP) transgenic mice that had been infected with adenovirus expressing Cre-recombinase to delete Smoothened showed over 75% knockdown of Smoothened. During culture, Smoothened-deficient LSEC had inhibited Hh signalling, less induction of capillarisation-associated genes and retention of fenestrae. In mice with injured livers, inhibiting Hh signalling prevented capillarisation. CONCLUSIONS: LSEC produce and respond to Hh ligands, and use Hh signalling to regulate complex phenotypic changes that occur during capillarisation. SN - 1468-3288 UR - https://www.unboundmedicine.com/medline/citation/22362915/Hedgehog_signalling_regulates_liver_sinusoidal_endothelial_cell_capillarisation_ L2 - http://gut.bmj.com/cgi/pmidlookup?view=long&pmid=22362915 DB - PRIME DP - Unbound Medicine ER -