In vivo inhibition of miR-155 significantly alters post-stroke inflammatory response.
In vivo inhibition of miR-155 significantly alters post-stroke inflammatory response.
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DOI：
10.1186/s12974-016-0753-x
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发表时间：
2016-11-09
影响因子：
9.3
通讯作者：
Roitbak T
中科院分区：
文献类型：
--
作者：
Pena-Philippides JC;Caballero-Garrido E;Lordkipanidze T;Roitbak T
MicroRNA miR-155参与各种病理条件下炎症过程的调节。在我们之前的研究中，我们证明了体内抑制miR-155促进小鼠实验性中风后的功能恢复。在本研究中，我们探讨了这种有益作用是否与miR-155抑制诱导的卒中后炎症反应改变有关。小鼠远端大脑中动脉闭塞（dMCAO）后48小时开始静脉注射特异性miR-155抑制剂。使用小鼠细胞因子基因和蛋白阵列和Western blot分析，在dMCAO后7、14和21天评估细胞因子和与细胞因子信号传导相关的关键分子表达的时间变化。采用电子和免疫荧光共聚焦显微镜技术评估dMCAO后不同时间点的超微结构变化，以及特定表型标记物表达的改变。在注射抑制剂的小鼠（抑制剂组）中，dMCAO后第7天CCL12和CXCL3细胞因子表达显著降低，第14天主要细胞因子IL-10、IL-4、IL-6、MIP-1α、IL-5和IL-17水平显著升高。这些时间变化与miR-155靶蛋白SOCS-1、SHIP-1和C/EBP-β的表达改变以及细胞因子信号调节因子STAT-3的磷酸化水平有关。电镜显示，抑制剂样品中具有吞噬活性的血管周围小胶质细胞/巨噬细胞数量减少。免疫荧光和Western blot显示，与对照组相比，dMCAO后第7天白细胞/巨噬细胞标志物CD45和吞噬标志物CD68的表达减少，而在第14天显著增加。基于我们的研究结果，我们提出小鼠脑卒中后体内miR-155抑制显著改变主要细胞因子和炎症相关分子表达的时间过程，从而影响实验性脑缺血后的炎症过程和组织修复。本文的在线版本（doi:10.1186/s12974-016-0753-x）包含补充材料，可供授权用户使用。
MicroRNA miR-155 is implicated in modulation of the inflammatory processes in various pathological conditions. In our previous studies, we demonstrated that in vivo inhibition of miR-155 promotes functional recovery after mouse experimental stroke. In the present study, we explored if this beneficial effect is associated with miR-155 inhibition-induced alterations in post-stroke inflammatory response. Intravenous injections of a specific miR-155 inhibitor were initiated at 48 h after mouse distal middle cerebral artery occlusion (dMCAO). Temporal changes in the expression of cytokines and key molecules associated with cytokine signaling were assessed at 7, 14, and 21 days after dMCAO, using mouse cytokine gene and protein arrays and Western blot analyses. Electron and immunofluorescence confocal microscopy techniques were used to evaluate the ultrastructural changes, as well as altered expression of specific phenotypic markers, at different time points after dMCAO. In the inhibitor-injected mice (inhibitor group), there was a significant decrease in CCL12 and CXCL3 cytokine expression at 7 days and significantly increased levels of major cytokines IL-10, IL-4, IL-6, MIP-1α, IL-5, and IL-17 at 14 days after dMCAO. These temporal changes correlated with altered expression of miR-155 target proteins SOCS-1, SHIP-1, and C/EBP-β and phosphorylation levels of cytokine signaling regulator STAT-3. Electron microscopy showed decreased number of phagocytically active peri-vascular microglia/macrophages in the inhibitor samples. Immunofluorescence and Western blot of these samples demonstrated that expression of leukocyte/ macrophage marker CD45 and phagocytosis marker CD68 was reduced at 7 days, and in contrast, significantly increased at 14 days after dMCAO, as compared to controls. Based on our findings, we propose that in vivo miR-155 inhibition following mouse stroke significantly alters the time course of the expression of major cytokines and inflammation-associated molecules, which could influence inflammation process and tissue repair after experimental cerebral ischemia. The online version of this article (doi:10.1186/s12974-016-0753-x) contains supplementary material, which is available to authorized users.
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