Linying Zhou, Wei Wang, Minxia Wu, Wenmin Zhang, Liangcheng Zhang
Department of Pathology, School of Basic Medical Sciences, Fujian Medical University, Fuzhou, China; Centre of Neuroscience, Fujian Medical University, Fuzhou, China.
Centre of Neuroscience, Fujian Medical University, Fuzhou, China.
Pediatr Neurol. 2014 Apr;50(4):357-62. doi: 10.1016/j.pediatrneurol.2013.12.007. Epub 2013 Dec 14.
Hypoxic-ischemic brain injury in neonates, especially in premature infants, is one of the main contributors to the mortality of newborns and can cause nervous system dysfunction in children. The major pathogenesis seems to be cerebral ischemia/reperfusion in the immature white matter that preferentially targets vulnerable premyelinating oligodendrocytes.
The goal of this study was to culture oligodendrocyte type 2 astrocyte cells in an oxygen and glucose deprivation environment to simulate ischemia injury and examine the cellular and molecular mechanisms involved in the neuroprotective effects of neuregulin-1ß on ischemia-induced immature oligodendrocytes.
Oligodendrocyte type 2 astrocyte cells were cultured from neonatal Sprague-Dawley rat cerebra. The cells were divided into two groups: one was subjected to oxygen and glucose deprivation for 9 hours and the other was treated with 50 ng/mL or 100 ng/mL neuregulin-1β during oxygen and glucose deprivation. Cell survival was determined by Trypan Blue staining and cell apoptosis were observed by fluorescein isothiocyanate-Annexin V and propidium iodide double staining. To study if the PI3K-Akt signaling pathway was involved in the mechanism of protective effect of neuregulin-1ß, Western blot analysis was used to quantitative the changes of protein.
Treatment with neuregulin-1ß within the period of oxygen and glucose deprivation significantly increased cell survival and also resulted in a significant decrease in cell apoptosis. The neuroprotective effects of neuregulin-1ß were prevented by treatment with Ly294002, an inhibitor of the phosphatidylinositol-3-kinase/Akt pathway.
These results suggest that neuregulin-1ß could protect the oligodendrocyte type 2 astrocyte progenitors against hypoxic injury, and the mechanism may be associated with the PI3K-Akt signaling pathway.
新生儿缺氧缺血性脑损伤,尤其是早产儿,是新生儿死亡的主要原因之一,可导致儿童神经系统功能障碍。主要发病机制似乎是未成熟白质中的脑缺血/再灌注,其优先靶向易损的少突胶质前体细胞。
本研究的目的是在氧和葡萄糖剥夺环境中培养少突胶质细胞2型星形胶质细胞,以模拟缺血损伤,并研究神经调节蛋白-1β对缺血诱导的未成熟少突胶质细胞神经保护作用的细胞和分子机制。
从新生Sprague-Dawley大鼠大脑中培养少突胶质细胞2型星形胶质细胞。将细胞分为两组:一组进行9小时的氧和葡萄糖剥夺,另一组在氧和葡萄糖剥夺期间用50 ng/mL或100 ng/mL神经调节蛋白-1β处理。通过台盼蓝染色测定细胞存活率,通过异硫氰酸荧光素-膜联蛋白V和碘化丙啶双重染色观察细胞凋亡。为了研究PI3K-Akt信号通路是否参与神经调节蛋白-1β的保护作用机制,采用蛋白质印迹分析法定量蛋白质的变化。
在氧和葡萄糖剥夺期间用神经调节蛋白-1β处理显著提高了细胞存活率,同时也导致细胞凋亡显著减少。磷脂酰肌醇-3-激酶/Akt通路抑制剂Ly294002处理可阻止神经调节蛋白-1β的神经保护作用。
这些结果表明,神经调节蛋白-1β可以保护少突胶质细胞2型星形胶质细胞祖细胞免受缺氧损伤,其机制可能与PI3K-Akt信号通路有关。