Tang Xiahong, Chen Feng, Lin Qinming, You Yan, Ke Jun, Zhao Shen
Department of Emergency, Fujian Provincial Hospital, Fujian Medical University, Fuzhou, Fujian 350009, P.R. China.
Fujian Provincial Institute of Emergency Medicien, Fuzhou, Fujian 350009, P.R. China.
Exp Ther Med. 2017 Nov;14(5):4312-4320. doi: 10.3892/etm.2017.5059. Epub 2017 Aug 28.
The present study aimed to investigate the beneficial effects and underlying mechanisms of bone marrow mesenchymal stem cells (BMSCs) on global ischemic hypoxic brain injury. Cells collected from the femurs and tibias of male Sprague Dawley rats were used to generate BMSCs following three culture passages. A rate model of cardiac arrest (CA) was induced by asphyxia. One hour following return of spontaneous circulation (ROSC), BMSCs were transplanted through injection into the tail vein. Neurological status was assessed using modified neurological severity score (mNSS) tests 1, 3 and 7 days following ROSC. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and immunohistochemical staining were used to detect insulin-like growth factor 1 (IGF-1) expression in the hippocampus. Furthermore, double-fluorescent labeling of green fluorescent protein (GFP) and IGF-1 was used to detect the IGF-1 expression in transplanted BMSCs. Serum levels of protein S100-B were examined using ELISA. GFP-labeled BMSCs were observed in the hippocampus at 1, 3 and 7 days post transplantation through fluorescent microscopy. BMSC transplantation resulted in reduced protein S100-B levels. The mNSS of the BMSC-treatment group was significantly reduced compared with that of the CA group. The RT-qPCR analysis and immunohistochemistry results demonstrated that BMSC treatment significantly increased IGF-1 expression in the hippocampus. In addition, the double-fluorescent labeling results demonstrated that transplanted BMSCs expressed IGF-1 in the hippocampus. The results of the present study suggest that BMSC treatment promotes the recovery of cerebral function following CA in rats possibly through the secretion of IGF-1.
本研究旨在探讨骨髓间充质干细胞（BMSCs）对全脑缺血缺氧性脑损伤的有益作用及潜在机制。从雄性Sprague Dawley大鼠的股骨和胫骨采集细胞，经三次传代培养后用于生成BMSCs。通过窒息诱导心脏骤停（CA）模型。自主循环恢复（ROSC）后1小时，经尾静脉注射移植BMSCs。在ROSC后1、3和7天，使用改良神经功能缺损评分（mNSS）测试评估神经功能状态。采用逆转录-定量聚合酶链反应（RT-qPCR）和免疫组化染色检测海马中胰岛素样生长因子1（IGF-1）的表达。此外，使用绿色荧光蛋白（GFP）和IGF-1的双荧光标记检测移植BMSCs中IGF-1的表达。采用酶联免疫吸附测定（ELISA）检测血清中蛋白S100-B的水平。通过荧光显微镜观察移植后1、3和7天海马中GFP标记的BMSCs。BMSC移植导致蛋白S100-B水平降低。与CA组相比，BMSC治疗组的mNSS显著降低。RT-qPCR分析和免疫组化结果表明，BMSC治疗显著增加了海马中IGF-1的表达。此外，双荧光标记结果表明，移植的BMSCs在海马中表达IGF-1。本研究结果表明，BMSC治疗可能通过分泌IGF-1促进大鼠CA后脑功能的恢复。
