摘要
目的探讨转染肝细胞生长因子(HGF)基因的骨髓间质细胞(MSCs)治疗大鼠缺血性脑卒中的效果。方法应用I型单纯疱疹病毒(HSV-1)载体经体外将HGF基因转染MSCs。制备鼠一过性大脑中动脉缺血模型,24 h后通过立体定向技术,分别将磷盐溶液(PBS)、转染MSCs和HGF基因的MSCs(MSC-HGF)移植入大鼠缺血脑组织。应用改良神经功能损伤严重性(mNSS)评分评价检测神经功能,应用2,3,5-氯化三苯基四氮唑染色法测量缺血脑组织容积。结果相同病毒感染滴度下,MSCs细胞培养上清液HGF浓度是MSCs细胞内的15倍,而且随感染滴度升高而显著增高(P<0.05)。治疗前各组大鼠mNSS评分无显著差异(P>0.05),治疗后4、7、14、21、28、35 d,MSC-HGF组较PBS组显著降低(P<0.05);治疗后14、21、28、35 d,MSC-HGF组较MSC组显著下降(P<0.05);治疗后14、21、28、35 d,MSC组较PBS组显著降低(P<0.05)。治疗后3 d,MSC-HGF组缺血脑组织容积较PBS组显著减少(P<0.05),PBS组和MSC组、MSC组和MSC-HGF组之间均无显著差异(P>0.05);治疗后14 d,MSC-HGF组缺血脑组织容积较PBS组和MSC均显著减小(P<0.05),MSC组较PBS组也显著减少(P<0.05)。结论 HGF基因经HSV-1载体体外转染MSCs可高效分泌HGF,转染HGF基因的MSCs治疗急性缺血性脑卒中有效,且较单纯MSCs治疗显著提高疗效。
Objective To observe the curative effect of hepatocyte growth factor （HGF）- transfected bone marrow stromal cells （MSCs） on cerebral uschemia. Methods HGF was transfected to MSCs in vitro by herpes simplex virus vector type one （HSV-1）. The models of cerebral ischemia were established were by transient middle cerebral artery occlusion （MACO） in 60 rats divided into 4 groups including phosphate-buffered saline （PBS） treatment group （n=18）, MSCs treatment group （n=18）, the green fluorescent protein-transfected MSCs （MSC-GFP） treatment group （n=6）, and HGF-transfected MSCs （MSC-HGF） treatment group （n=18）. All the agents were stereotactically transplanted into the ischemic cerebral areas 24 hours after transient MCAO. The curative effects including the neurological function recovery and cerebral ischemic area volume, which were determined respectively by the behavioral test and 2, 3, 5-triphenyhetrazolium-chloride （TTC） staining, were compared among all the groups. Results Behavioral tests showed that the neurological deficits were significantly improved in MSC-HGF treatment group compared with the other three groups （P〈0.05）. TTC staining showed that the volume of the cerebral ischemic area was significant decrease in MSC-HGF treatment group compared with both PBS and MSC group 2 weeks after the treatment （P〈0.05）. Conclusions MSC-HGF may be made vitro by HSV-1. The present study suggested that the curative effect of MSCs-HGF on cerebral ischemia is better than that of MSCs.
出处
《中国临床神经外科杂志》
2014年第7期414-417,共4页
Chinese Journal of Clinical Neurosurgery
基金
上海市重点学科建设项目(ZK2010A31)
脑卒中多学科一体化救治学科群建设项目(PWZxkq2011-02)
关键词
缺血性脑卒中
肝细胞生长因子
骨髓间质细胞
Ⅰ型单纯疱疹病毒
脑内移植
Cerebral ischemia
Hepatocyte growth factor
Bone marrow stromal cells
Herpes simplex virus vector type one
Intracerebral transplantation
