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Gene expression profiling of brain cortex microvessels may support brain vasodilation in acute liver failure rat models

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Abstract

Development of brain edema in acute liver failure can increase intracranial pressure, which is a severe complication of the disease. However, brain edema is neither entirely cytotoxic nor vasogenic and the specific action of the brain microvasculature is still unknown. We aimed to analyze gene expression of brain cortex microvessels in two rat models of acute liver failure. In order to identify global gene expression changes we performed a broad transcriptomic approach in isolated brain cortex microvessels from portacaval shunted rats after hepatic artery ligation (HAL), hepatectomy (HEP), or sham by array hybridization and confirmed changes in selected genes by RT-PCR. We found 157 and 270 up-regulated genes and 143 and 149 down-regulated genes in HAL and HEP rats respectively. Western blot and immunohistochemical assays were performed in cortex and ELISA assays to quantify prostaglandin E metabolites were performed in blood of the sagittal superior sinus. We Identified clusters of differentially expressed genes involving inflammatory response, transporters-channels, and homeostasis. Up-regulated genes at the transcriptional level were associated with vasodilation (prostaglandin-E synthetase, prostaglandin-E receptor, adrenomedullin, bradykinin receptor, adenosine transporter), oxidative stress (hemoxygenase, superoxide dismutase), energy metabolism (lactate transporter) and inflammation (haptoglobin). The only down-regulated tight junction protein was occludin but slightly. Prostaglandins levels were increased in cerebral blood with progression of liver failure. In conclusion, in acute liver failure, up-regulation of several genes at the level of microvessels might suggest an involvement of energy metabolism accompanied by cerebral vasodilation in the cerebral edema at early stages.

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Abbreviations

ALF:

Acute liver failure

BBB:

Blood-Brain-Barrier

BSA:

Bovine serum albumin

CBF:

Cerebral blood flow

Ct:

Cycle threshold

Fc:

Fold-change

HAL:

Hepatic artery ligation

HEP:

Hepatectomy

HRP:

Horseradish peroxidase

IV:

Intravenous

PCA:

Portacaval anastomosis

PGEM:

Prostaglandin E metabolites

RT-PCR:

Real time reverse transcription polymerase chain reaction

TJ:

Tight junction

ICP:

Intracranial pressure

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Acknowledgments

The authors want to give special thanks to their mentor Prof. Cordoba, deceased in 2014. We will always remember him for his honesty, generosity and friendship. Thanks for showing us the best way to do science.

Author contributions

Study concept and design: Ll. P., M.O., J.C.

Acquisition of data: Ll. P., M.O., J. R-G. T. G-L, L.C.

Analysis and interpretation of data: Ll. P., L.C., M.O., J.C.

Drafting of the manuscript: Ll. P., M.O., L.C., J.C.

Critical revision of the manuscript for important intellectual content: J.C.

Statistical analysis: Ll. P., M.O., J.C.

Obtention of funding: J.C., M.O.

Technical or material support: J. R-G, T. G-L.

Study supervision: J.C.

Author information

Authors and Affiliations

  1. Servei de Medicina Interna-Hepatologia, Valld’Hebron Institut de Recerca (VH-IR), Barcelona, Spain

    Lluis Palenzuela, Marc Oria, Jordi Romero-Giménez, Teresa Garcia-Lezana, Laia Chavarria & Juan Cordoba

  2. Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Instituto de Salud Carlos III, Madrid, Spain

    Lluis Palenzuela, Marc Oria, Teresa Garcia-Lezana, Laia Chavarria & Juan Cordoba

  3. Departament de Medicina, Universitat Autònoma de Barcelona, Barcelona, Spain

    Lluis Palenzuela, Marc Oria, Jordi Romero-Giménez, Teresa Garcia-Lezana, Laia Chavarria & Juan Cordoba

  4. Center for Fetal, Cellular and Mollecular Therapy, Division of Pediatric General and Thoracic Surgery, Cincinnati Children’s Hospital Medical Center (CCHMC), Cincinnati, OH, USA

    Marc Oria

Authors
  1. Lluis Palenzuela
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  2. Marc Oria
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  3. Jordi Romero-Giménez
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  4. Teresa Garcia-Lezana
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  5. Laia Chavarria
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  6. Juan Cordoba
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Corresponding author

Correspondence to Marc Oria.

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Disclosures

The authors have nothing to disclose.

Grant support

Grant support by Fundació Investigació Sant Pau (FISP), Barcelona and Sheila Sherlock EASL Post-Doc Fellowship to Marc Oria, PhD.

Study supported by Fondo de Investigaciones Sanitarias PI08/0698, PI11/0954 co-financed by the European Regional Development Fund (ERDP).

CIBERehd is supported by Instituto de Salud Carlos III, Madrid, Spain.

Additional information

Lluis Palenzuela and Marc Oria contributed equally to the work

Marc Oria and Juan Cordoba have equal responsibility for all aspects of this publication

Juan Cordoba died in January 27, 2014

Electronic supplementary material

Supplementary Fig. 1

Brain Cortex Microvessels a) Extracted microvessels (MV) were smeared and observed under an optical microscope. Magnification 40×. b) Enrichment was assessed by Real Time PCR for the endothelial marker GLUT1 (GIF 1133 kb)

High resolution image (TIFF 36203 kb)

Supplementary Fig. 2

RTime PCR in Sham and PCA rats for Ptger4, Ptges, Bdkrb2 and Adm genes. Expression levels for the listed genes were normalized for β actin (GIF 92 kb)

High resolution image (TIFF 2480 kb)

Supplementary Fig. 3

Ptges and Bdkrb2 Western Blot. Protein levels detected for Ptges, Bdkrb2 in SHAM, HAL and HEP samples (GIF 512 kb)

High resolution image (TIFF 13491 kb)

Supplementary Data 1

(DOCX 124 kb)

Supplementary Table 1

(DOCX 17 kb)

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Palenzuela, L., Oria, M., Romero-Giménez, J. et al. Gene expression profiling of brain cortex microvessels may support brain vasodilation in acute liver failure rat models. Metab Brain Dis 31, 1405–1417 (2016). https://doi.org/10.1007/s11011-016-9863-5

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  • DOI: https://doi.org/10.1007/s11011-016-9863-5

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