Astrocytic endothelin-1 overexpression promotes neural progenitor cells proliferation and differentiation into astrocytes via the Jak2/Stat3 pathway after stroke
Astrocytic endothelin-1 overexpression promotes neural progenitor cells proliferation and differentiation into astrocytes via the Jak2/Stat3 pathway after stroke
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星形胶质细胞内皮素-1过表达促进中风后神经祖细胞增殖并通过Jak2/Stat3途径分化为星形胶质细胞
DOI：
10.1186/s12974-019-1597-y
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发表时间：
2019-11-16
影响因子：
9.3
通讯作者：
Chung, Sookja K.
中科院分区：
文献类型：
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作者：
Cheng, Xiao;Yeung, Patrick K. K.;Chung, Sookja K.
背景内皮素-1（ET-1）是脑卒中时星形胶质细胞合成和表达上调的一种蛋白质。我们以前证明过表达星形胶质细胞ET-1（GET-1）的转基因小鼠表现出更严重的神经功能缺损，其特征是短暂性大脑中动脉闭塞（tMCAO）后出现更大的梗死。ET-1是一种已知的血管收缩剂、促有丝分裂剂和存活因子。然而，目前还不清楚是否在GET-1小鼠中风后观察到严重的脑损伤是由于ET-1神经发生失调，通过改变干细胞niche.MethodsNon-transgenic（Ntg）和GET-1小鼠进行tMCAO与1小时闭塞，然后长期再灌注（从第1天到第28天）。采用四点量表法评估神经功能。2，3，5-三苯基四氮唑氯化物染色测定细胞面积和体积。采用溴脱氧尿苷（BrdU）、Ki 67和Sox 2、Nestin和Doublecortin（DCX）免疫荧光双标记法检测神经干细胞（NSC）在脑室下区（SVZ）的增殖和迁移。采用BrdU和神经元特异性核蛋白（NeuN）、BrdU和胶质细胞酸性蛋白（GFAP）免疫荧光双标染色评价SVZ中NSC的分化。磷酸化Stat 3（p-Stat 3）的表达通过Western-blot和免疫荧光染色检测。结果GET-1小鼠在tMCAO损伤后表现出更严重的神经功能缺损和更大的梗死面积。在tMCAO后28天，GET-1脑同侧SVZ处BrdU标记的祖细胞增殖显著增加，其与GFAP共表达。在tMCAO后7天，Ntg和GET-1小鼠缺血脑中的p-Stat 3表达增加。在tMCAO后7天，GET-1脑中同侧的p-Stat 3表达显著高于Ntg脑中的p-Stat 3表达。此外，GET-1小鼠AG 490（JAK 2/Stat 3抑制剂）治疗显示了显着减少神经功能缺损沿着减少梗死面积和矮化星形胶质细胞分化后tMCAO.ConclusionsThe数据表明，星形胶质细胞内皮素-1过度表达促进祖细胞干细胞增殖和星形胶质细胞分化通过JAK 2/Stat 3途径。
BackgroundEndothelin-1 (ET-1) is synthesized and upregulated in astrocytes under stroke. We previously demonstrated that transgenic mice over-expressing astrocytic ET-1 (GET-1) displayed more severe neurological deficits characterized by a larger infarct after transient middle cerebral artery occlusion (tMCAO). ET-1 is a known vasoconstrictor, mitogenic, and a survival factor. However, it is unclear whether the observed severe brain damage in GET-1 mice post stroke is due to ET-1 dysregulation of neurogenesis by altering the stem cell niche.MethodsNon-transgenic (Ntg) and GET-1 mice were subjected to tMCAO with 1 h occlusion followed by long-term reperfusion (from day 1 to day 28). Neurological function was assessed using a four-point scale method. Infarct area and volume were determined by 2,3,5-triphenyltetra-zolium chloride staining. Neural stem cell (NSC) proliferation and migration in subventricular zone (SVZ) were evaluated by immunofluorescence double labeling of bromodeoxyuridine (BrdU), Ki67 and Sox2, Nestin, and Doublecortin (DCX). NSC differentiation in SVZ was evaluated using the following immunofluorescence double immunostaining: BrdU and neuron-specific nuclear protein (NeuN), BrdU and glial fibrillary acidic protein (GFAP). Phospho-Stat3 (p-Stat3) expression detected by Western-blot and immunofluorescence staining.ResultsGET-1 mice displayed a more severe neurological deficit and larger infarct area after tMCAO injury. There was a significant increase of BrdU-labeled progenitor cell proliferation, which co-expressed with GFAP, at SVZ in the ipsilateral side of the GET-1 brain at 28 days after tMCAO. p-Stat3 expression was increased in both Ntg and GET-1 mice in the ischemia brain at 7 days after tMCAO. p-Stat3 expression was significantly upregulated in the ipsilateral side in the GET-1 brain than that in the Ntg brain at 7 days after tMCAO. Furthermore, GET-1 mice treated with AG490 (a JAK2/Stat3 inhibitor) sh owed a significant reduction in neurological deficit along with reduced infarct area and dwarfed astrocytic differentiation in the ipsilateral brain after tMCAO.ConclusionsThe data indicate that astrocytic endothelin-1 overexpression promotes progenitor stem cell proliferation and astr ocytic differentiation via the Jak2/Stat3 pathway.