SPARC expression by cerebral microvascular endothelial cells in vitro and its influence on blood-brain barrier properties.
SPARC expression by cerebral microvascular endothelial cells in vitro and its influence on blood-brain barrier properties.
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DOI：
10.1186/s12974-016-0657-9
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发表时间：
2016-08-31
影响因子：
9.3
通讯作者：
Quandt JA
中科院分区：
文献类型：
--
作者：
Alkabie S;Basivireddy J;Zhou L;Roskams J;Rieckmann P;Quandt JA
分泌蛋白（分泌的酸性和富含半胱氨酸的蛋白）是一种非结构性的细胞基质调节蛋白，参与血管生成和内皮屏障功能，但其在脑血管发育、炎症和中枢神经系统（CNS）修复中的潜在作用尚未确定。本研究检测了培养的人脑微血管内皮细胞（hCMEC/D3）-血脑屏障（BBB）的体外模型-在增殖和屏障表型之间转换并遇到促炎刺激时的表达。通过Western印迹和免疫细胞化学和RT-PCR的信使RNA（mRNA）的定量蛋白质水平。随着细胞成熟并建立汇合单层，增殖的hCMEC/D3 s的组成型表达减少。PCNA表达与增殖标志物Ki-67呈正相关，提示PCNA在脑血管发育中的作用。促炎分子肿瘤坏死因子-α（TNF-α）和内毒素脂多糖（LPS）可增加脑内皮细胞中TNF α的表达。干扰素γ（IFN-γ）消除TNF-α单独给药时观察到的TNF-α诱导。屏障功能测定显示重组人（rh）-β-D-半乳糖苷酶增加细胞旁通透性和降低跨内皮电阻（TEER）。与含有生理剂量的rh-Lactobacillus的培养基中的细胞相比，暴露于rh-Lactobacillus（1-10 μg/ml）的hCMEC/D3中的闭合小带-1（ZO-1）和闭合蛋白表达减少。总之，这些发现定义了在BBB的发育和炎症过程中影响脑微血管特性和功能的作用，使得它可以介导CNS炎症和修复的过程。本文的在线版本（doi：10.1186/s12974-016-0657-9）包含补充材料，可供授权用户使用。
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. 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. 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. 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. The online version of this article (doi:10.1186/s12974-016-0657-9) contains supplementary material, which is available to authorized users.
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影响因子：
3.2
作者：
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通讯作者：
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影响因子：
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DOI：
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作者：
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