神经干细胞通过P25和P35蛋白质促进缺血缺氧性脑病新生大鼠神经功能恢复  

Effects of Neural Stem Cells on Treating Hypoxic-Ischemic Encephalopathy through P25 and P35 Proteins

作　　者：田雨光[1] 庞炜[1] 安雷 魏伟 顾为望[1] 王玉珏[1] 岳敏[1] 

机构地区：[1]南方医科大学实验动物中心暨比较医学研究所,广州510515 [2]南方医科大学第三附属医院神经外科,广州510630 [3]广东省脐带血造血干细胞库,广州510000

出　　处：《中国细胞生物学学报》2017年第11期1435-1440,共6页Chinese Journal of Cell Biology

基　　金：广东省自然科学基金(批准号:2017A030313192);国家自然科学基金(批准号:801402625);广东省医学科学技术研究基金(批准号:A2017124;A2016255);广东省科技计划项目(批准号:2016A030303008;2017A030303019)资助的课题~~

摘　　要：该文探讨了神经干细胞(neural stem cells,NSCs)对缺血缺氧性脑病(hypoxic-ischemic encephalopathy,HIE)新生大鼠的治疗作用及P25、P35蛋白质水平变化的影响。60只新生7 d SD大鼠随机分为3组,每组20只。Sham组:施假手术;模型组:HIE造模,以PBS处理;治疗组:HIE造模,以NSCs治疗。干预后第5、7、14、21 d作神经行为学检测。第21 d处死,取组织切片进行HE染色作病理分析。Western blot法检测海马区P25、P35蛋白质水平。免疫荧光双染验证P25、P35蛋白质水平。Rotarod test结果显示,与Sham组比较,模型组与治疗组大鼠在转轮中坚持时间明显下降(P<0.05);造模后第7、14、21 d,治疗组大鼠在转轮中坚持时间升高趋势明显高于模型组(P<0.05)。Morris水迷宫结果显示,与模型组相比,治疗组大鼠的逃避潜伏期在各个时间点均明显下降(P<0.05)。模型组大鼠出现海马区神经元水肿、核染色质结构不清、空泡形成等病理变化,治疗组病理损伤明显减轻。Western blot结果显示,与Sham组相比,模型组P35水平减少,P25水平增加(P<0.05);治疗组与模型组相比较P35水平增多,P25水平减少(P<0.05)。免疫荧光双染在细胞学水平验证了P35、P25水平变化。该研究结果表明,NSCs可促进大鼠缺血脑损伤后神经功能恢复,可能通过上调P35蛋白质水平,下调P25蛋白质水平,对脑缺血损伤起保护作用。The objective of this study is to evaluate the efficacy on neurological functional recovery after transplantation of neural stem cells （NSCs） and the changes of P25 and P35 proteins in adult rats. 60 rats （7-day-old Sprague-Dawley） were randomly assigned to sham-operated group （n=20）, ischemia group （n=20） and NSCs transplanted group （n=20）. Rotarod test and Morris test were performed at 5, 7, 14, 21 days after treatment to measure the behaviors of the rats. Hematoxylin-eosin staining was used to observe the pathological changes of brain damage in neonatal rats. Westem blot was used to analyse the levels of P35 and P25 in hippocampus. The irnmunofluorescence double dye was used to verify the levels of P35 and P25 proteins. Rotarod test result showed that motor function recovered gradually and restored to near the normal level on the 21th day in NSCs group and there was a significant difference between NSCs group and HIE group on the 14th day （P〈0,05）. Morris test showed that escape latency significantly shortened in NSCs group at each time points compared with HIE group （P〈0.05）. Pathological results showed that hippocampal neuron edema, nuclear chromatin structure is not clear, and vacuoles appeared in HIE group. Compared with HIE group, pathological changes in brain were significantly lessened. Western blot showed that the changes of P35 and P25 levels were significantly different in other groups compared with the Sham group. The P35 level decreased while P25 level increased in HIE group （P〈0.05）. Compared with the HIE group, the P35 levels increased while P25 level decreased in NSCs group （P〈0.05）. The levels of P35 and P25 protein were verified by immunofluoreseence double dye in cytological level. Our results showed that NSCs promoted the recovery of neurological function in rats after ischemic brain injury possibly by up regulating P35 level expression and decreasing the expression of P25 protein level.

关 键 词：缺血性脑病 神经干细胞 P35 P25 神经功能 

分 类 号：R742[医药卫生—神经病学与精神病学]