【摘要】 目的观察三维(3-D)球体间充质干细胞(MSCs)移植对脑缺血再灌注损伤大鼠脑组织肿瘤坏死因子α(TNF-α)及凋亡相关蛋白表达的影响,探讨3-D球体MSCs的神经保护作用及可能机制。方法实验大鼠随机分为假手术组、溶剂对照组、3-D球体MSCs治疗组(n=36),假手术组常规分离大脑中动脉(MCA)但不结扎,MSCs治疗组采用线栓法建立大脑中动脉缺血(MCAO)模型后给予MSCs移植,溶剂对照组模型建立成功后给予等体积溶剂对照。各组大鼠分别于移植治疗后1、3、7d进行神经功能评分;采用ELISA和RT-PCR方法检测大鼠脑组织中的TNF-α表达;采用免疫组化SABC法染色,观察损伤侧大鼠脑组织Caspase-3和Caspase-8蛋白的表达。结果与假手术组、溶剂对照组相比,MSCs治疗组大鼠神经运动功能评分降低(P<0.05);与假手术组相比,溶剂对照组大鼠脑组织TNF-α、Caspase-3和Caspase-8表达增加(P<0.01);与溶剂对照组相比,MSCs治疗组脑组织TNF-α、Caspase-3、Caspase-8表达下降(P<0.05,P<0.01)。结论 3-D球体MSCs移植可能通过下调脑组织中炎性因子TNF-α含量及减少凋亡相关蛋白的表达,改善大鼠脑缺血再灌注损伤后神经运动功能。更多还原

【Abstract】 Objective To investigate the effect of the placenta-derived three-dimensional(3-D)spheroid-cultured mesenchymal stem cells(MSCs)on the expression of tumor necrosis factor-α(TNF-α)and apoptosis-related proteins in rats with cerebral ischemia-reperfusion injury,so as to explore the neuroprotective effects and mechanism of 3-D spheroid-cultured MSCs.Methods The experimental animals were randomly divided into Sham group,Vehicle group and 3-D spheroid-cultured MSCs treated group(n=36).In the Sham group,the middle cerebral artery(MCA)was simply isolated but not ligated.In the MSC treated group,focal ischemia/reperfusion in rats was induced by intraluminal middle cerebral artery occlusion(MCAO)with a nylon monofilament suture and then MSCs transplantation was done.In the Vehicle group,the MCAO model was set up and an equal volume of vehicle was given.Neurological scores of animals were evaluated at different time points:the first day,the third day and the seventh day after surgery.Enzyme-linked immunosorbent assay(ELISA)and RT-PCR method were used to detect TNF-αexpression in rat brain tissues.The SABC immunohistochemical staining method was used to detect the expression of Caspase-3and Caspase-8in rat brain tissues.Results The neuromotor function of rats in MSCs treated group was significantly decreased compared with those in the other two groups(P <0.05).Compared with the Sham group,the expressions of TNF-α,Caspase-3and Caspase-8in the brain tissue were significantly increased in the Vehicle group(P<0.01).Moreover,compared with the Vehicle group,expression of TNF-α,Caspase-3 and Caspase-8in the brain tissue was significantly decreased in MSCs treated group(P<0.05,P<0.01).Conclusion 3-D spheroid-cultured MSCs transplantation may improve the neuromotor function in rats with cerebral ischemia/reperfusion injury via down-regulating inflammatory cytokine TNF-αand expression of Caspase-3and Caspase-8in the brain tissue.更多还原