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Journal of Neuroinflammation

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Open Access 01-12-2016 | Research

SPARC expression by cerebral microvascular endothelial cells in vitro and its influence on blood-brain barrier properties

Authors: Samir Alkabie, Jayasree Basivireddy, Lixin Zhou, Jane Roskams, Peter Rieckmann, Jacqueline A. Quandt

Published in: Journal of Neuroinflammation | Issue 1/2016

Abstract

Background

SPARC (secreted protein acidic and rich in cysteine) is a nonstructural, cell-matrix modulating protein involved in angiogenesis and endothelial barrier function, yet its potential role in cerebrovascular development, inflammation, and repair in the central nervous system (CNS) remains undetermined.

Methods

This study examines SPARC expression in cultured human cerebral microvascular endothelial cells (hCMEC/D3)—an in vitro model of the blood-brain barrier (BBB)—as they transition between proliferative and barrier phenotypes and encounter pro-inflammatory stimuli. SPARC protein levels were quantified by Western blotting and immunocytochemistry and messenger RNA (mRNA) by RT-PCR.

Results

Constitutive SPARC expression by proliferating hCMEC/D3s is reduced as cells mature and establish a confluent monolayer. SPARC expression positively correlated with the proliferation marker Ki-67 suggesting a role for SPARC in cerebrovascular development. The pro-inflammatory molecules tumor necrosis factor-α (TNF-α) and endotoxin lipopolysaccharide (LPS) increased SPARC expression in cerebral endothelia. Interferon gamma (IFN-γ) abrogated SPARC induction observed with TNF-α alone. Barrier function assays show recombinant human (rh)-SPARC increased paracellular permeability and decreased transendothelial electrical resistance (TEER). This was paralleled by reduced zonula occludens-1 (ZO-1) and occludin expression in hCMEC/D3s exposed to rh-SPARC (1–10 μg/ml) compared with cells in media containing a physiological dose of SPARC.

Conclusions

Together, these findings define a role for SPARC in influencing cerebral microvascular properties and function during development and inflammation at the BBB such that it may mediate processes of CNS inflammation and repair.

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Title: SPARC expression by cerebral microvascular endothelial cells in vitro and its influence on blood-brain barrier properties
Authors: Samir Alkabie
Jayasree Basivireddy
Lixin Zhou
Jane Roskams
Peter Rieckmann
Jacqueline A. Quandt
Publication date: 01-12-2016
Publisher: BioMed Central
Published in: Journal of Neuroinflammation / Issue 1/2016
Electronic ISSN: 1742-2094
DOI: https://doi.org/10.1186/s12974-016-0657-9

Keynote webinar | Spotlight on medication adherence

Springer Medicine

SPARC expression by cerebral microvascular endothelial cells in vitro and its influence on blood-brain barrier properties

Abstract

Background

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

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