Amplification and propagation of interleukin-1β signaling by murine brain endothelial and glial cells
Amplification and propagation of interleukin-1β signaling by murine brain endothelial and glial cells
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DOI：
10.1186/s12974-017-0908-4
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发表时间：
2017-07-01
影响因子：
9.3
通讯作者：
Marks, Daniel L.
中科院分区：
文献类型：
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作者：
Krasnow, Stephanie M.;Knoll, J. Gabriel;Marks, Daniel L.
背景资料：在急性感染和慢性疾病期间，促炎细胞因子白细胞介素-1 β（IL-1 β）在大脑内起作用，引起代谢紊乱和疾病行为。目前尚不清楚大脑中哪些细胞是IL-1 β产生这些疾病反应的近端靶点。我们进行了一系列体外实验，以（1）研究哪些脑细胞群体对IL-1 β表现出炎症反应，以及（2）检查各种共培养组合中不同IL-1 β反应细胞类型之间的相互作用。我们用PBS或IL-1 β处理小鼠脑微血管内皮细胞（BMEC）、星形胶质细胞和小胶质细胞的原代培养物，然后进行qPCR以测量炎症基因表达或免疫细胞化学以评估活化B细胞的核因子κ轻链增强子（NF-κ B）活化。为了评估星形胶质细胞和/或BMEC是否向小胶质细胞传播炎症信号，我们将小胶质细胞暴露于星形胶质细胞条件培养基，并在transwell中共培养内皮细胞和胶质细胞。治疗组进行了比较学生的t检验或方差分析，然后由Bonferroni校正的t tests.Results：IL-1 β增加炎症基因的表达和NF-κ B激活的主要鼠混合神经胶质细胞，丰富的星形胶质细胞，BMEC文化。尽管IL-1 β引起炎症基因表达的最小变化，并且在分离的小胶质细胞中不诱导NF-κ B的核转位，但是当与星形胶质细胞和/或BMEC共培养时，这些细胞被IL-1 β更稳健地激活。我们观察到一个极化的内皮细胞对IL-1 β的反应，因为应用IL-1 β的近腔内皮表面产生了一个更复杂的小胶质细胞的炎症反应比发生以下管腔IL-1 β exposation.Conclusions：炎症信号检测，放大，并通过中枢神经系统通过一个连续的和混响的信号级联涉及脑内皮细胞和神经胶质细胞之间的通信传播。我们提出，大脑的先天免疫反应不同，这取决于哪一侧的血脑屏障的炎症刺激的出现，从而使大脑作出不同的反应，中央与外周炎症的侮辱。
Background: During acute infections and chronic illnesses, the pro-inflammatory cytokine interleukin-1 beta (IL-1 beta) acts within the brain to elicit metabolic derangements and sickness behaviors. It is unknown which cells in the brain are the proximal targets for IL-1 beta with respect to the generation of these illness responses. We performed a series of in vitro experiments to (1) investigate which brain cell populations exhibit inflammatory responses to IL-1 beta and (2) examine the interactions between different IL-1 beta-responsive cell types in various co-culture combinations.Methods: We treated primary cultures of murine brain microvessel endothelial cells (BMEC), astrocytes, and microglia with PBS or IL-1 beta, and then performed qPCR to measure inflammatory gene expression or immunocytochemistry to evaluate nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kappa B) activation. To evaluate whether astrocytes and/or BMEC propagate inflammatory signals to microglia, we exposed microglia to astrocyte-conditioned media and co-cultured endothelial cells and glia in transwells. Treatment groups were compared by Student's t tests or by ANOVA followed by Bonferroni-corrected t tests.Results: IL-1 beta increased inflammatory gene expression and NF-kappa B activation in primary murine-mixed glia, enriched astrocyte, and BMEC cultures. Although IL-1 beta elicited minimal changes in inflammatory gene expression and did not induce the nuclear translocation of NF-kappa B in isolated microglia, these cells were more robustly activated by IL-1 beta when co-cultured with astrocytes and/or BMEC. We observed a polarized endothelial response to IL-1 beta, because the application of IL-1 beta to the abluminal endothelial surface produced a more complex microglial inflammatory response than that which occurred following luminal IL-1 beta exposure.Conclusions: Inflammatory signals are detected, amplified, and propagated through the CNS via a sequential and reverberating signaling cascade involving communication between brain endothelial cells and glia. We propose that the brain's innate immune response differs depending upon which side of the blood-brain barrier the inflammatory stimulus arises, thus allowing the brain to respond differently to central vs. peripheral inflammatory insults.