Protective Effect of Ad-VEGF-Bone Mesenchymal Stem Cells on Cerebral Infarction
Protective Effect of Ad-VEGF-Bone Mesenchymal Stem Cells on Cerebral Infarction
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DOI：
10.5137/1019-5149.jtn.11488-14.3
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发表时间：
2016-01-01
影响因子：
0.8
通讯作者：
Lan, Qing
中科院分区：
文献类型：
--
作者：
Chen, Bo;Zhang, Feng;Lan, Qing
目的：为探讨血管内皮生长因子（VEGF）基因修饰的骨髓间充质干细胞（BMSCs）脑室内移植治疗脑梗死的机制，采用Zea-Longa线栓法建立大鼠大脑中动脉闭塞缺血再灌注（MCAO I/R）模型。重组腺病毒（Ad-VEGF）被设计成表达VEGF。将大鼠分为3组。在大脑中动脉闭塞（MCAO）后24 h，将对照腺病毒感染的BMSC（BMSC-Ad）、Ad-VEGF感染的BMSC（BMSC-Ad-VEGF）和磷酸盐缓冲盐水（PBS）悬液分别注射到第1、2和3组大鼠的侧脑室内。采用改良的神经功能严重度评分（mNSS）评价大鼠的神经功能。用氯化三苯基四氮唑（TTC）染色法测定脑梗死体积。免疫组化法检测缺血半暗带GFAP和pGSK 3 β的表达。GFAP、pAKT、AKT和pGSK3 β的表达用Western blot测定。结果：接受BMSC-Ad的动物的功能改善加速，而在MCAO后7天至28天的所有时间内，移植BMSC-Ad-VEGF的动物的功能改善显著大于其他治疗的动物。结论：PI3K/AKT/GSK3 β信号通路可减少大鼠脑缺血后反应性胶质增生，改善神经功能缺损，减少脑梗死体积，促进血管新生。
AIM: To understand the mechanism of intracerebroventricular transplantation of vascular endothelial growth factor (VEGF) gene modified bone mesenchymal stem cells (BMSCs) in rats after cerebral infarction.MATERIAL and METHODS: The middle cerebral artery occlusion ischemia/reperfusion (MCAO I/R) model was established in rats using the Zea-Longa suture method. A recombinant adenovirus (Ad-VEGF) was engineered to express VEGF. The rats were divided into 3 groups. Control BMSC infected with control adenovirus (BMSC-Ad), BMSC infected by Ad-VEGF (BMSC-Ad-VEGF), and phosphate buffered saline (PBS) suspension were injected into the intracerebroventricular system of the rats in groups 1, 2 and 3 respectively, 24 hours after middle cerebral artery occlusion (MCAO). The neurological function of rats was evaluated with the modified Neurological Severity Scores (mNSS). The infarct volume of brain in rats was determined using 2,3,5-triphenyltetrazolium chloride (TTC) stain at 14 days. GFAP and pGSK3 beta expression of ischemic penumbra was determined using immunohistochemical method. GFAP, pAKT, AKT, and pGSK3 beta expressions were determined with Western blot.RESULTS: Functional improvement was accelerated in animals receiving BMSC-Ad, while improvement at all times between 7 days and 28 days post MCAO was significantly greater in animals transplanted with BMSC-Ad-VEGF than for other treated animals. The number of GFAP-labeled cells was prevented by post-ischemic BMSC-Ad-VEGF treatment; pMCAO activate the PI3K/AKT/GSK3 beta pathway to reduce reactive gliosis.CONCLUSION: Our findings demonstrate that PI3K/AKT/GSK3 beta pathway could reduce reactive gliosis, ameliorate neurological deficit, diminish the percentage of cerebral infarction volume in rats, and facilitate angiogenesis.