Transplantation of neural stem cells overexpressing glial cell line-derived neurotrophic factor enhances Akt and Erk1/2 signaling and neurogenesis in rats after stroke
Transplantation of neural stem cells overexpressing glial cell line-derived neurotrophic factor enhances Akt and Erk1/2 signaling and neurogenesis in rats after stroke
复制标题
过表达神经胶质细胞系源性神经营养因子的神经干细胞移植可增强中风后大鼠的 Akt 和 Erk1/2 信号传导和神经发生
DOI：
10.3760/cma.j.issn.0366-6999.20122965
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发表时间：
2013-04-05
影响因子：
6.1
通讯作者：
Yuan Qiong-lan
中科院分区：
文献类型：
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作者：
Yuan Miao;Wen Sheng-jun;Yuan Qiong-lan
背景我们之前的研究表明，移植过度表达胶质细胞系衍生神经营养因子（GDNF）的神经干细胞（NSC）对中风后的大鼠具有有益作用。然而，其背后的机制存在很大争议。在本研究中，我们探讨了神经发生、Akt和细胞外信号调节激酶1/2（Erk1/2）信号是否参与了这一过程。方法通过阻断大脑中动脉2小时并再灌注诱导短暂性缺血性卒中。再灌注后3天，给予GDNF/NSC、NSC和载体。免疫组织化学染色用于评估巢蛋白抗体的神经发生；通过Western blotting分析检测Akt和Erk1/2的磷酸化。结果再灌注后1至7周，GDNF/NSCs和NSCs移植与对照组（vehicle）相比，巢蛋白阳性细胞显着增加，并且在再灌注后2和3周，GDNF/NSCs比NSCs表现出更强的效果。同时，与对照组相比，GDNF/NSCs组和NSCs组的Erk1/2磷酸化水平均升高，并且GDNF/NSCs组的Erk1/2磷酸化水平在任何给定时间均显着高于NSCs组。相反，与对照组相比，GDNF/NSCs 组和 NSCs 组中丝裂原激活蛋白激酶磷酸酶-1 (MKP-1)（称为 Erk1/2 信号抑制剂）的表达显着降低。此外，与对照组和NSCs组相比，GDNF/NSCs组的Akt磷酸化水平显着增强和延长。 结论 移植GDNF/NSCs可增强神经发生并激活Akt和Erk1/2信号传导，这可能为GDNF/NSCs在中风治疗中提供潜力。
Background Our previous studies have indicated that the beneficial effects of grafting neural stem cells (NSCs) overexpressing glial cell line-derived neurotrophic factor (GDNF) in rats after stroke. However, the underlying mechanisms are highly debatable. In this study, we investigated whether neurogenesis, Akt, and extracellular signal-regulated kinase 1/2 (Erk1/2) signaling were involved in this process.Methods Transient ischemic stroke were induced by occluding middle cerebral artery for 2 hours and reperfusion. At 3 days after reperfusion, GDNF/NSCs, NSCs, and vehicle were administered. Immunohistochemical staining was used to evaluate neurogenesis by nestin antibody; phosphorylation of Akt and Erk1/2 was investigated by Western blotting analysis.Results Transplantation of GDNF/NSCs and NSCs significantly increased nestin-positive cells compared to control group (vehicle) from 1 to 7 weeks after reperfusion, and GDNF/NSCs showed stronger effect than NSCs at 2 and 3 weeks after reperfusion. Meanwhile, enhanced phosphorylation level of Erk1/2 was observed in the GDNF/NSCs and NSCs groups compared with control group, and phosphorylation level of Erk1/2 in GDNF/NSCs group was remarkably higher than that of NSCs group at any given time. In contrast, expression of mitogen-activated protein kinase phosphatase-1 (MKP-1), known as inhibitor of Erk1/2 signaling, was significantly decreased in the GDNF/NSCs and NSCs groups compared with the control group. Moreover, much enhanced and prolonged phosphorylation level of Akt of GDNF/NSCs group was detected compared with control and NSCs group.Conclusion Grafting GDNF/NSCs enhances neurogenesis and activates Akt and Erk1/2 signaling, that may provide the potential for GDNF/NSCs in stroke treatment.