Zhu Jun-de, Wang Jun-Jie, Ge Guo, Kang Chao-Sheng
Department of Anatomy, School of Basic Medicine, Guizhou Medical University, Guian New Area, China.
Department of Anatomy, School of Basic Medicine, Guizhou Medical University, Guian New Area, China.
J Stroke Cerebrovasc Dis. 2017 Jul;26(7):1547-1559. doi: 10.1016/j.jstrokecerebrovasdis.2017.02.034. Epub 2017 May 3.
To investigate neuroprotection of noggin-transfected neural stem cells (NSCs) against focal cerebral ischemia reperfusion injury (IRI) in rats.
Eighty Wistar rats were randomly divided into the sham, IRI, NSCs, and noggin + NSCs groups. Noggin containing adenoviral vectors was transfected into rat NSCs. Rats were subjected to 2.0 hours middle cerebral artery occlusion and reperfusion 1.0 hour, followed by infusion into the lateral ventricles of NSCs alone, noggin-transfected NSCs, and saline at 3 days in the NSCs, noggin + NSCs, and sham groups, respectively. All rats were sacrificed on 1, 3, 7, and 28 days after transplantation; the colorimetric method was used to detect the levels of superoxide dismutase (SOD) and the malondialdehyde (MDA) content after the behavior capability determined. Western blot was performed for detecting the expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) proteins. The TUNEL-positive and BrdU/nestin double-positive cells were observed under a light microscope and quantitative analysis was performed by morphometric technique.
Noggin-transfected NSCs significantly decreased the infarct volume and improved the neurological scores. Noggin-transfected NSCs also reduced the percentage of apoptotic neurons and relieved neuronal morphological damage. Noggin-transfected NSC transplantation markedly decreased the MDA levels and increased the SOD activity, and simultaneously downregulated the BMP4 (bone morphogenesis protein), VEGF, and bFGF proteins.
The present study demonstrates that grafting NSCs modified by noggin gene provides better neuroprotection for cerebrovascular disease.
探讨经诺金蛋白(noggin)转染的神经干细胞(NSCs)对大鼠局灶性脑缺血再灌注损伤(IRI)的神经保护作用。
将80只Wistar大鼠随机分为假手术组、IRI组、NSCs组和noggin + NSCs组。将携带诺金蛋白基因的腺病毒载体转染至大鼠神经干细胞。大鼠接受2.0小时大脑中动脉闭塞及1.0小时再灌注,然后分别于术后3天向NSCs组、noggin + NSCs组和假手术组大鼠侧脑室注入单纯神经干细胞、经诺金蛋白转染的神经干细胞及生理盐水。于移植后1、3、7和28天处死所有大鼠;在测定行为能力后,采用比色法检测超氧化物歧化酶(SOD)水平及丙二醛(MDA)含量。采用蛋白质免疫印迹法检测血管内皮生长因子(VEGF)和碱性成纤维细胞生长因子(bFGF)蛋白的表达。在光学显微镜下观察TUNEL阳性细胞及BrdU/巢蛋白双阳性细胞,并采用形态计量学技术进行定量分析。
经诺金蛋白转染的神经干细胞显著减小梗死体积并改善神经功能评分。经诺金蛋白转染的神经干细胞还降低了凋亡神经元的比例并减轻了神经元形态学损伤。经诺金蛋白转染的神经干细胞移植显著降低了MDA水平并提高了SOD活性,同时下调了骨形态发生蛋白4(BMP4)、VEGF和bFGF蛋白的表达。
本研究表明,移植经诺金蛋白基因修饰的神经干细胞可为脑血管疾病提供更好的神经保护作用。