Fingolimod effects in neuroinflammation: Regulation of astroglial glutamate transporters?
Fingolimod effects in neuroinflammation: Regulation of astroglial glutamate transporters?
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DOI：
10.1371/journal.pone.0171552
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Linker RA
中科院分区：
文献类型：
--
作者：
Lee DH;Seubert S;Huhn K;Brecht L;Rötger C;Waschbisch A;Schlachetzki J;Klausmeyer A;Melms A;Wiese S;Winkler J;Linker RA
芬戈莫德是一种口服鞘氨醇-1-磷酸受体调节剂，可减少免疫细胞的再循环，也可直接靶向胶质细胞。本研究探讨了芬戈莫德对促炎条件下星形胶质细胞谷氨酸转运体表达的影响。在星形胶质细胞培养中，促炎细胞因子的加入导致谷氨酸转运体谷氨酸转运体-1 （slc1a2/ slc1a2）和谷氨酸天冬氨酸转运体（slc1a3/ slc1a3）在mRNA或蛋白水平上的表达显著下调。在这种情况下，直接应用于星形胶质细胞的fingolimod-1 phosphate （F1P）未改变slc1a2和slc1a3 mRNA的表达水平。Western Blot和免疫细胞化学对两种转运蛋白在蛋白水平上的分析也未发现F1P的任何作用。在功能水平上，从F1P处理的星形胶质细胞中添加条件上清液到神经元细胞培养中并没有导致神经突生长增加。在作为多发性硬化症模型的实验性自身免疫性脑脊髓炎中，芬戈莫德治疗减少了T细胞和巨噬细胞/小胶质细胞介导的炎症，也减少了星形胶质细胞的激活。同时，fingolimod在mRNA水平上恢复了slc1a2和slc1a3在炎症脊髓中的表达，在蛋白水平上恢复了slc1a2和slc1a3的表达，可能是通过间接的抗炎机制。这些发现为该化合物在神经炎症中的主要外周作用提供了进一步的证据。
Fingolimod is an oral sphingosine-1-phosphate-receptor modulator which reduces the recirculation of immune cells and may also directly target glial cells. Here we investigate effects of fingolimod on expression of astroglial glutamate transporters under pro-inflammatory conditions. In astrocyte cell culture, the addition of pro-inflammatory cytokines led to a significant downregulation of glutamate transporters glutamate transporter-1 (slc1a2/SLC1A2) and glutamate aspartate transporter (slc1a3/SLC1A3) expression on the mRNA or protein level. In this setting, the direct application of fingolimod-1 phosphate (F1P) on astrocytes did not change expression levels of slc1a2 and slc1a3 mRNA. The analysis of both transporters on the protein level by Western Blot and immunocytochemistry did also not reveal any effect of F1P. On a functional level, the addition of conditioned supernatants from F1P treated astrocytes to neuronal cell culture did not result in increased neurite growth. In experimental autoimmune encephalomyelitis as a model of multiple sclerosis, fingolimod treatment reduced T cell and macrophages/microglia mediated inflammation and also diminished astrocyte activation. At the same time, fingolimod restored the reduced expression of slc1a2 and slc1a3 in the inflamed spinal cord on the mRNA level and of SLC1A2 and SLC1A3 on the protein level, presumably via indirect, anti-inflammatory mechanisms. These findings provide further evidence for a predominantly peripheral effect of the compound in neuroinflammation.