Liang Jingjing, Cui Ronghui, Wang Jinglei, Shen Jiabing, Chen Ying, Cao Maosheng, Ke Kaifu
Department of Neurology, Affiliated Hospital of Nantong University, Nantong, China.
Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong, China.
Front Neurol. 2021 Feb 4;12:613547. doi: 10.3389/fneur.2021.613547. eCollection 2021.
Skin-derived Precursor Schwann cells (SKP-SCs) have been reported to provide neuroprotection for the injured and dysmyelinated nervous system. However, little is known about SKP-SCs on acute ischemic stroke (AIS). We aimed to explore the efficacy and the potential mechanism of action of SKP-SCs on AIS in a rat ischemic stroke model. Adult male Sprague-Dawley rats were subjected to a middle cerebral artery occlusion (MCAO) for 1.5 h on Day 0 and subsequently received an intracarotid injection of 2 × 10 green fluorescent protein (GFP) -labeled SKP-SCs or phosphate buffered saline (PBS) during reperfusion. Neurological function was assessed by behavioral tests on Days 1, 4, 7, 14, and 28. In a satellite cohort, rat brains were harvested and infarct volume was measured with 2,3,5-triphenyltetrazolium chloride (TTC) staining on Days 1 and 7, and migration and survival of SKP-SCs in the brain were traced by monitoring green fluorescence at 6 and12 h on Day 0, and on Days 1, 4, 7, 14, and 28. Histopathology and immunofluorescence staining were used to analyze the morphology, survival and apoptosis of neurons. Additionally, in an SKP-SC co-culture model using fetal rat primary cortical neurons underwent oxygen glucose deprivation/reoxygenation (OGD/R), Western blot was used to detect the expression of apoptosis indicators including activated caspase-3, Bax, and Bcl-2. TUNEL staining was used to count apoptotic cells. Intracarotid transplantation of SKP-SCs effectively migrated to the periinfarct area and survived for at least 4 weeks. Transplanted SKP-SCs inhibited neuronal apoptosis, reduced infarct volume, and improved neurological recovery in the MCAO rats. Moreover, data showed that SKP-SCs treatment inhibited OGD/R-induced neuronal apoptosis and promoted survival of the cultured primary cortical neurons. Intracarotid transplantation of SKP-SCs promoted functional recovery in the rat AIS model and possesses the potential to be further developed as a novel therapy to treat ischemic stroke in humans.
皮肤源性前体雪旺细胞（SKP-SCs）已被报道可为受损和脱髓鞘的神经系统提供神经保护作用。然而，关于SKP-SCs在急性缺血性中风（AIS）方面的研究却知之甚少。我们旨在探讨SKP-SCs对大鼠缺血性中风模型中AIS的疗效及潜在作用机制。成年雄性Sprague-Dawley大鼠于第0天接受大脑中动脉闭塞（MCAO）1.5小时，随后在再灌注期间经颈内动脉注射2×10绿色荧光蛋白（GFP）标记的SKP-SCs或磷酸盐缓冲盐水（PBS）。在第1、4、7、14和28天通过行为测试评估神经功能。在一个卫星队列中，于第1天和第7天收获大鼠大脑，用2,3,5-三苯基四氮唑氯化物（TTC）染色测量梗死体积，并在第0天以及第1、4、7、14和28天通过监测绿色荧光追踪SKP-SCs在大脑中的迁移和存活情况。采用组织病理学和免疫荧光染色分析神经元的形态、存活及凋亡情况。此外，在使用经历氧糖剥夺/复氧（OGD/R）的胎鼠原代皮质神经元的SKP-SC共培养模型中，采用蛋白质免疫印迹法检测包括活化的半胱天冬酶-3、Bax和Bcl-2在内的凋亡指标的表达。采用TUNEL染色对凋亡细胞进行计数。经颈内动脉移植的SKP-SCs有效迁移至梗死周边区域并存活至少4周。移植的SKP-SCs抑制了MCAO大鼠的神经元凋亡，减小了梗死体积，并改善了神经功能恢复。此外，数据表明SKP-SCs治疗可抑制OGD/R诱导的神经元凋亡并促进培养的原代皮质神经元存活。经颈内动脉移植SKP-SCs可促进大鼠AIS模型的功能恢复，具有进一步开发成为治疗人类缺血性中风新疗法的潜力。
