Activation of c-Jun N-terminal kinase and p38 after cerebral ischemia upregulates cerebral sodium-glucose transporter type 1
Activation of c-Jun N-terminal kinase and p38 after cerebral ischemia upregulates cerebral sodium-glucose transporter type 1
复制标题
脑缺血后 c-Jun N 末端激酶和 p38 的激活上调脑钠葡萄糖转运蛋白 1 型
DOI：
10.1016/j.ejphar.2017.02.007
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Tokuyama S
中科院分区：
文献类型：
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作者：
Yamazaki Y.;Arita K.;Harada S.;Tokuyama S
我们最近报道脑钠-葡萄糖转运蛋白1（SGLT-1）在脑缺血加重中起作用。然而，大脑SGLT-1的作用机制仍不清楚。在这里，我们证明了通过脑SGLT-1的钠内流加剧了脑缺血性神经元损伤。使用α-甲基-D-吡喃葡萄糖苷（α-MG）诱导SGLT特异性钠离子内流。原代皮层神经元细胞内的钠浓度估计使用钠结合苯并呋喃三乙酸荧光。使用SGLT-1 siRNA在原代皮质神经元和小鼠中实现SGLT-1敲低。在处理后1天，使用生化测定评估原代培养的皮层神经元的存活率。使用大脑中动脉闭塞（MCAO）在SGLT-1敲低小鼠中产生局灶性脑缺血模型。在MCAO后1天评估空腹血糖水平、梗死发展和行为异常的变化。用200 mM α-MG处理诱导细胞内钠浓度持续增加，并且这种增加在α-MG去除后恢复正常。神经元SGLT-1敲低对100 µM H2 O2诱导的神经元细胞死亡没有影响;然而，敲低阻止了17.5 mM葡萄糖和100 µM H2 O2/8.75 mM葡萄糖联合处理诱导的神经元细胞死亡。神经元SGLT-1敲低也抑制了α-MG单独和100 µM H2 O2/0.01 mM α-MG联合处理诱导的细胞死亡。我们的体内结果表明，脑SGLT-1敲低小鼠侧脑室给予5.0 µg α-MG/小鼠诱导的脑缺血性神经元损伤加重得到改善。因此，通过脑SGLT-1的钠内流可能加剧脑缺血诱导的神经元损伤。
We recently reported that cerebral sodium-glucose transporter type 1 (SGLT-1) plays a role in exacerbation of cerebral ischemia. However, the mechanism by which cerebral SGLT-1 acts remains unclear. Here we demonstrated that sodium influx through cerebral SGLT-1 exacerbates cerebral ischemic neuronal damage. SGLT-specific sodium ion influx was induced using α-methyl-D-glucopyranoside (α-MG). Intracellular sodium concentrations in primary cortical neurons were estimated using sodium-binding benzofuran isophthalate fluorescence. SGLT-1 knockdown in primary cortical neurons and mice was achieved using SGLT-1 siRNA. The survival rates of primary cultured cortical neurons were assessed using biochemical assays 1 day after treatment. Middle cerebral artery occlusion (MCAO) was used to generate a focal cerebral ischemic model in SGLT-1 knockdown mice. The change in fasting blood glucose levels, infarction development, and behavioral abnormalities were assessed 1 day after MCAO. Treatment with 200 mM α-MG induced a continuous increase in the intracellular sodium concentration, and this increase was normalized after α-MG removal. Neuronal SGLT-1 knockdown had no effect on 100 µM H2O2-induced neuronal cell death; however, the knockdown prevented the neuronal cell death induced by 17.5 mM glucose and the co-treatment of 100 µM H2O2/8.75 mM glucose. Neuronal SGLT-1 knockdown also suppressed the cell death induced by α-MG alone and the co-treatment of 100 µM H2O2/0.01 mM α-MG. Ourin vivoresults showed that the exacerbation of cerebral ischemic neuronal damage induced by the intracerebroventricular administration of 5.0 µg α-MG/mouse was ameliorated in cerebral SGLT-1 knockdown mice. Thus, sodium influx through cerebral SGLT-1 may exacerbate cerebral ischemia-induced neuronal damage.