Department of Neurosurgery, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China.
Cell Biochem Biophys. 2013 Jul;66(3):671-80. doi: 10.1007/s12013-012-9511-6.
This study aimed to investigate the expression of the Nemo-like kinase (NLK) in the brain after experimental subarachnoid hemorrhage (SAH) in rats. A total of 90 rats were randomly divided into six groups: control group, day 1, day 3, day 5, day 7, and day 14. Day 1, day 3, day 5, day 7, and day 14 groups were all SAH groups in which the rats were killed on days 1, 3, 5, 7, and 14, respectively. In SAH groups, autologous arterial blood was injected into cisterna magna once on day 0. Cross-sectional area of basilar artery was measured by H&E staining. Immunostaining and immunoblotting experiments were performed to detect the expression of NLK protein. Real-time polymerase chain reaction was used to analyze the presence and quantity of NLK mRNA. The level of oxidative stress in the artery was also measured. The basilar arteries exhibited vasospasm after SAH and became the most severe on day 3. The expressions of NLK protein and mRNA were decreased remarkably in SAH groups compared with the control group. The down-regulated expression of NLK was detected after SAH and the low ebb was on day 3, which was oppositely the peak time of oxidative stress. The expression of NLK was present mainly in the neurons in the brain and smooth muscle cells in the basilar artery. NLK is decreasingly expressed in an opposite time-course to the development of cerebral vasospasm (CVS) and SAH-induced brain injury in this rat experimental model of SAH and these findings might have important implications during the administration of specific NLK agonist to prevent or reduce CVS or neuronal apoptosis caused by SAH.
本研究旨在探讨神经母细胞瘤样激酶(NLK)在大鼠实验性蛛网膜下腔出血(SAH)后脑内的表达。90 只大鼠随机分为 6 组:对照组、第 1 天、第 3 天、第 5 天、第 7 天和第 14 天。第 1 天、第 3 天、第 5 天、第 7 天和第 14 天组均为 SAH 组,大鼠分别于第 1、3、5、7 和 14 天处死。SAH 组于第 0 天一次性向脑池内注入自体动脉血。用 H&E 染色测量基底动脉横截面积。进行免疫染色和免疫印迹实验检测 NLK 蛋白表达。实时聚合酶链反应分析 NLK mRNA 的存在和数量。还测量了动脉中的氧化应激水平。SAH 后基底动脉发生血管痉挛,第 3 天最为严重。与对照组相比,SAH 组 NLK 蛋白和 mRNA 的表达显著降低。SAH 后 NLK 的下调表达在第 3 天检测到,低谷出现在第 3 天,与氧化应激的高峰期相反。NLK 的表达主要存在于大脑中的神经元和基底动脉中的平滑肌细胞中。NLK 在大鼠 SAH 实验模型中,其表达随脑动脉痉挛(CVS)和 SAH 诱导的脑损伤的发展呈相反的时间进程减少,这些发现可能对特异性 NLK 激动剂的给药具有重要意义,以预防或减少 CVS 或由 SAH 引起的神经元凋亡。