骨髓单个核细胞移植对脑出血大鼠神经修复方面的影响

发布时间：2018-05-25 04:12

本文选题：大鼠 + 脑出血　；参考：《郑州大学》2014年硕士论文

【摘要】：研究背景脑出血是临床上常见的脑血管疾病，具有高致死率和高致残率的特点，脑出血后出血损伤中心区域的神经细胞死亡，进而导致相应的神经功能缺失，至今尚无真正有效的治疗措施，目前移植具有自我更新和定向分化能力的外源性的多潜能干细胞是脑出血最具潜能的治疗措施，骨髓单个核细胞因其具有提取容易、无需培养及可自体移植等诸多优点，使之成为治疗急性期脑出血的最佳种子细胞之一，最新研究证明静脉移植BMMNCs安全有效，且能够在特定的微环境的诱导下定向分化为多种细胞。目的探讨骨髓单个核细胞（BMMNCs）移植治疗对脑出血大鼠的胶质源性神经营养因子（GDNF）的表达、其脑内神经元样细胞和星形胶质样细胞分化情况及神经功能恢复的影响。方法纹状体区立体定向注射胶原酶建立脑出血模型，筛选成功模型后随机分为模型组（ICH组）和BMMNCs移植组（BMMNCs组），BMMNCs组于造模后24h后经尾静脉注入1mL含3×107个BMMNCs细胞悬液，分别于术后1d、7d、14d、28d采用改良的大鼠神经功能评分（mNSS）观察大鼠神经行为学表现；于1d、7d、14d采用Western blotting方法检测脑内GDNF的表达情况；28d后采用免疫组织荧光双标法示踪BMMNCs脑内分化情况。结果术后7d、14d、28d，BMMNCs组mNSS评分均明显低于ICH组，差异有统计学意义（P0.05）；两组的GDNF表达均于1d开始出现，7d达高峰，14d逐渐下降，ICH组各时间点GDNF的表达量均明显低于BMMNCs组，差异有统计学意义（P0.05）；术后28d，，BMMNCs组纹状体区可检测到Brdu和NeuN双标阳性细胞及Brdu和GFAP双标阳性细胞。结论 BMMNCs移植能显著改善大鼠神经行为学功能，显著增加GDNF的分泌量，并能够在脑出血大鼠体内分化为神经元样细胞和星形胶质样细胞。
[Abstract]:Research background Intracerebral hemorrhage (ICH) is a common cerebrovascular disease with high mortality and high disability rate. So far, there is no effective treatment. Transplantation of exogenous pluripotent stem cells with self-renewal and directional differentiation is the most potential treatment for intracerebral hemorrhage, and bone marrow mononuclear cells are easy to extract because of their ability of self-renewal and directional differentiation. It is one of the best seed cells for the treatment of acute intracerebral hemorrhage without the advantages of culture and autologous transplantation. The latest research has proved that vein transplantation BMMNCs is safe and effective. And can be induced by specific microenvironment to differentiate into a variety of cells. Purpose To investigate the effects of bone marrow mononuclear cell transplantation (BMMNCs) on the expression of glial derived neurotrophic factor (GDNF) in rats with intracerebral hemorrhage (ICH), the differentiation of neuron-like cells and astrocytes in the brain and the recovery of neural function. Method The model of intracerebral hemorrhage was established by stereotactic injection of collagenase in the striatum. After screening, the model group was randomly divided into two groups: the model group (ICH group) and the BMMNCs transplantation group (BMMNCs group). After 24 hours, 1mL was injected into the caudal vein with 3 脳 107 BMMNCs cell suspensions. The neurobehavioral manifestations of rats were observed by modified rat neural function score (mNSSs) at 1 day, 7 days and 14 days after operation, and the expression of GDNF in brain was detected by Western blotting method on day 1 and day 7, respectively. After 28 days, the differentiation of BMMNCs in brain was traced by immunofluorescence double labeling method. Result The mNSS scores in the BMMNCs group were significantly lower than those in the ICH group (P 0.05), the GDNF expression in both groups began to appear on the 1st day and reached the peak on the 1st day and gradually decreased on the 14th day after operation. The expression of GDNF at all time points in the ICH group was significantly lower than that in the BMMNCs group. Brdu and NeuN double labeled positive cells and Brdu and GFAP double labeled positive cells could be detected in striatum of BMMNCs group 28 days after operation. Conclusion BMMNCs transplantation could significantly improve the neurobehavioral function of rats, increase the secretion of GDNF, and differentiate into neuron-like cells and astrocyte-like cells in rats with intracerebral hemorrhage.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R743.34

【参考文献】