Department of Traumatic Surgery& Emergency Surgery, The 1st Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang,325000, China.
Department of Ward 233, The 1st Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang,325000, China.
Cell Mol Biol (Noisy-le-grand). 2023 Sep 30;69(9):125-129. doi: 10.14715/cmb/2023.69.9.18.
The aim of this experiment was to analyze the ameliorating effect of neural stem cells (NSCs) on focal cerebral ischemia (FCI) through GDNF/PI3K/AKT axis, so as to provide evidence for future clinical application of NSCs. In this study, the 15 Sprague-Dawley (SD) male rats were modeled for middle cerebral artery occlusion (MCAO)-induced FCI and then grouped: NSCs group was treated with NSC transplantation, GDNF/NSCs group was transplanted with recombinant adenovirus pAdEasy-1-pAdTrackCMV-GDNF-transfedcted NSCs, and the blank group was treated with normal saline transplantation. Rats were tested by rotarod and corner turn tests at 1 week and 4 weeks after NSC transplantation, and the levels of tumor necrosis factor-α (TNF-α), interleukin-6/8 (IL-6/8), superoxide dismutase (SOD) and malondialdehyde (MDA) were quantified. Then all rats were killed and their brain tissues were HE stained for the determination of and GDNF/PI3K/AKT axis-associated protein expression. The results of the experiment showed that: at the 1st and 4th week after transplantation, the time on the rod, number of turnings and SOD were the lowest in the blank group among the three groups, while IL-6, IL-8, TNF-α and MDA were the highest (P<0.05). Increased time on the rod, number of turnings and SOD, as well as decreased IL-6, IL-8, TNF-α and MDA were observed in NSCs and GDNF/NSCs groups after transplantation, with better performance in GDNF/NSCs group (P<0.05). Based on HE staining of brain tissue, GDNF/NSCs group had the most significant improvement in tissue injury and the highest GDNF, PI3K, AKT and p-AKT protein expression among the three groups (P<0.05). In conclusions, NSC transplantation can ameliorate neurological function in MCAO-induced FCI rats through the GDNF/PI3K/AKT axis.
本实验旨在通过 GDNF/PI3K/AKT 轴分析神经干细胞（NSCs）对局灶性脑缺血（FCI）的改善作用，为 NSCs 的临床应用提供依据。本研究对 15 只 Sprague-Dawley（SD）雄性大鼠进行大脑中动脉闭塞（MCAO）诱导的 FCI 造模，然后分组：NSCs 组进行 NSC 移植，GDNF/NSCs 组移植转染重组腺病毒 pAdEasy-1-pAdTrackCMV-GDNF 的 NSCs，空白组进行生理盐水移植。在 NSC 移植后 1 周和 4 周，通过转棒和转角测试评估大鼠的运动能力，同时定量检测肿瘤坏死因子-α（TNF-α）、白细胞介素-6/8（IL-6/8）、超氧化物歧化酶（SOD）和丙二醛（MDA）的水平。然后处死所有大鼠，对其脑组织进行 HE 染色，以确定 GDNF/PI3K/AKT 轴相关蛋白的表达。实验结果表明：在移植后的第 1 周和第 4 周，三组中空白组大鼠在棒上的时间、转弯次数和 SOD 最低，而 IL-6、IL-8、TNF-α和 MDA 最高（P<0.05）。NSCs 和 GDNF/NSCs 组移植后，在棒上的时间、转弯次数和 SOD 增加，IL-6、IL-8、TNF-α和 MDA 减少，其中 GDNF/NSCs 组效果更好（P<0.05）。基于脑组织的 HE 染色，三组中 GDNF/NSCs 组的组织损伤改善最显著，GDNF、PI3K、AKT 和 p-AKT 蛋白表达最高（P<0.05）。综上所述，NSC 移植可通过 GDNF/PI3K/AKT 轴改善 MCAO 诱导的 FCI 大鼠的神经功能。
