胎盘来源3D球体间充质干细胞对大鼠脑缺血/再灌注损伤Caspase-3的影响

【作者】 张明；

【导师】 夏玉军；

【作者基本信息】 青岛大学 ， 人体解剖与组织胚胎学（专业学位）， 2015， 硕士

【摘要】 研究背景:脑梗塞,是一种由于大脑血管堵塞而导致脑缺血性发作的疾病,已经严重危害到人类的健康。血栓溶解疗法和脑保护疗法已经应用于临床治疗脑梗塞。但是随着脑部科学的发展与探索和神经再生技术领域的研究拓展,细胞疗法和细胞替代疗法已经成为近年来的热点。本研究旨在探讨间充质干细胞在脑缺血再灌注大鼠的神经功能恢复和caspase-3在神经细胞表达的影响。方法:建立大鼠脑梗死模型。治疗组注射2106脐带血间充质干细胞;假手术组和对照组注入相同数量的培养基。在注射治疗之前和注射后1天,3天,7天分别进行神经功能评分。Caspase-3mRNA和蛋白的表达水平的检测分别使用RT-PCR法和免疫组织化学法。细胞凋亡的分析评估用TUNEL工具包。结果:被标记过得脐带血间充质干细胞注入大鼠后治疗组大鼠的神经功能恢复相比于对照组有明显改善。RT-PCR和免疫组织化学的实验结果表明caspase-3的表达水平与对照组相比较有显著地减少,而假手术组和对照组没有显著差异。细胞凋亡结果表明治疗组和对照组相比,TUNEL检测的阳性细胞会有减少。结论:脐带血来源的间充质干细胞能够促进脑缺血再灌注大鼠的神经功能恢复并且减少caspase-3在神经细胞中的表达。更多还原

【Abstract】 Background: Cerebral infarction,a type of ischemic stroke disease resulting from a blockage in the blood vessels supplying blood to the brain, is a threat to human health. Thrombolytic therapy and brain protection methods have been used to treat cerebral infarction clinically. But with the development of Brain Science and nerve regeneration, cell therapy and cell replacement therapy become a hotspot in recent years. This study aimed to explore the effect of placenta-derived mesenchymal stem cells on the neurological function of cerebral ischemia-reperfusion rats and the expression of caspase-3 in nerve cells.Methods: The rat cerebral infarction models were established. The treatment group was injected 2×106 Umbilical Cord Blood Mesenchymal Stem Cells;Sham-operated group and the control were injected into the same amount of cell culture media. Evaluation of neurological function was performed before and after the injection: 1day, 3days and 7days. The mRNA and protein expression level of caspase-3 was detected using RT-PCR and immunohistochemical method, respectively. Apoptosis analysis was assessed using a TUNEL kit.Results:Labeled Umbilical Cord Blood Mesenchymal Stem Cells survived well after being injected into rats; The neurological function of treatment rats improved compared with the control(F=29.996,t=2.236,n=6,p<0.05). RT-PCR and immunohistochemistry results indicated that the expression level of caspase-3 decreased in treatment rats(F=116.502~466.271, t=7.208~23.168, P<0.01), while there was no significant difference between the sham-operated group and the control(P>0.05). Apoptosis results showed that TUNEL-positive cells were significantly lower in the treatment group than the control group.Conclusions: This study indicated that placenta-derived mesenchymal stem cells contribute to the recovery of neurological function of cerebral ischemia-reperfusion rats and the expression of caspase-3 decreased in nerve cells.更多还原