Chu Kon, Jung Keun-Hwa, Kim Se-Jeong, Lee Soon-Tae, Kim Juhyun, Park Hee-Kwon, Song Eun-Cheol, Kim Seung U, Kim Manho, Lee Sang Kun, Roh Jae-Kyu
Stroke and Neural Stem Cell Laboratory in Clinical Research Institute, Department of Neurology, Seoul National University Hospital, Seoul, South Korea.
Brain Res. 2008 May 1;1207:182-92. doi: 10.1016/j.brainres.2008.02.043. Epub 2008 Mar 4.
Hypoxia-inducible factor-1 (HIF-1) plays important roles in the prevention of cerebral ischemia. Deferoxamine (DFX), an iron chelator stabilizes the HIF-1alpha and activates target genes involved in compensation for ischemia. In this study, we are to investigate whether HIF-1alpha can be stabilized in human neural stem cells (NSCs) by DFX, and pre-transplantation of NSCs with HIF-1alpha stabilization can induce prolonged ischemic tolerance. In the DFX-treated NSCs, the HIF-1alpha protein expression was increased about 100-fold time-dependently, and subsequent transcriptional activation (VEGF, BDNF and CXCR4) was also observed. To test an ability to induce ischemic prevention in vivo, DFX-treated NSCs or naïve NSCs were transplanted in the striatum of adult rats. Seven days following the transplantation, focal cerebral ischemia was done. Infarct volumes were reduced in both NSCs-transplanted groups, compared with ischemia-only, but more reduced in DFX-treated NSCs group. The protective effects of NSCs were ablated when HIF-1alpha was silenced. HIF-1alpha protein levels were increased in both NSCs-transplanted groups, but more increased in DFX-treated NSCs group. RT-PCR analysis manifested a downregulation of mRNA expression of TNF-alpha, IL-6 and MMP-9 in both NSCs groups, but further decrease in DFX-treated NSCs group. These findings provide evidence that HIF-1alpha stabilization in human NSCs can be achieved effectively by DFX, and HIF-1alpha-stabilized NSCs protect against ischemia in a preventive mode.
缺氧诱导因子-1(HIF-1)在预防脑缺血中发挥着重要作用。去铁胺(DFX),一种铁螯合剂,可稳定HIF-1α并激活参与缺血代偿的靶基因。在本研究中,我们旨在探究DFX是否能在人神经干细胞(NSCs)中稳定HIF-1α,以及预先移植经HIF-1α稳定处理的NSCs是否能诱导延长的缺血耐受性。在经DFX处理的NSCs中,HIF-1α蛋白表达呈时间依赖性增加约100倍,随后还观察到转录激活(VEGF、BDNF和CXCR4)。为了测试在体内诱导缺血预防的能力,将经DFX处理的NSCs或未处理的NSCs移植到成年大鼠的纹状体中。移植后7天,进行局灶性脑缺血。与仅缺血组相比,两个NSCs移植组的梗死体积均减小,但经DFX处理的NSCs组减小得更多。当HIF-1α沉默时,NSCs的保护作用消失。两个NSCs移植组中HIF-1α蛋白水平均升高,但经DFX处理的NSCs组升高得更多。RT-PCR分析表明,两个NSCs组中TNF-α、IL-6和MMP-9的mRNA表达均下调,但经DFX处理的NSCs组进一步降低。这些发现提供了证据,表明DFX能有效实现人NSCs中HIF-1α的稳定,且经HIF-1α稳定处理的NSCs以预防模式保护免受缺血损伤。