移植脑源性神经生长因子修饰骨髓间充质干细胞对帕金森病大鼠模型治疗作用的实验研究

发布时间：2018-10-16 21:24

【摘要】：目的:观察移植pIRESneo-EGFP-BDNF修饰骨髓间充质干细胞(mesenchymal stem cells,MSCs)对帕金森病(Parkinson’sdisease,PD)大鼠的研究。方法 :将pEGFP(N1)-BDNF质粒进行改造与pIRESneo相连结,构建高拷贝质粒pIRESneo-EGFP-BDNF,采用电穿孔法转染骨髓MSCs;制备PD大鼠模型,随机分为假手术组、PD组、MSCs组、脑源性神经生长因子(brain-derivedneurotrophic factor,BDNF)组,经侧脑室移植MSCs或pIRESneo-EGFP-BDNF修饰骨髓MSCs术后2、4、8周,腹腔注射阿朴吗啡诱导PD大鼠旋转行为;采用Western blot方法测定中脑黑质BDNF、酪氨酸羟化酶(tyrosine hydroxylase,TH)蛋白的含量。结果:双酶切鉴定pIRESneo-EGFP-BDNF构建成功;移植术后2、4、8周MSCs组和BDNF组大鼠与PD组比较旋转次数明显减少(P0.05),以BDNF组较MSCs比较改善更为明显(P0.05);移植细胞干预PD模型8周后中脑黑质BDNF、TH蛋白的含量,BDNF组较MSCs组、PD组明显增高(P0.05)。结论:经侧脑室移植pIRESneo-EGFP-BDNF修饰的骨髓MSCs能明显改善PD大鼠的行为能力,显著提高中脑黑质BDNF、TH蛋白的含量。
[Abstract]:Aim: to observe the effects of pIRESneo-EGFP-BDNF modified bone marrow mesenchymal stem cells (mesenchymal stem cells,MSCs) on Parkinson's disease (Parkinson'sdisease,PD) rats. Methods: pEGFP (N1)-BDNF plasmid was modified with pIRESneo to construct the PD rat model of high copy plasmid pIRESneo-EGFP-BDNF, transfected with bone marrow MSCs; by electroporation. PD rats were randomly divided into sham-operation group, PD group, MSCs group, and brain derived nerve growth factor (brain-derivedneurotrophic factor,BDNF) group. The rotation behavior of PD rats induced by apomorphine was induced by intraperitoneal injection of apomorphine and the content of BDNF, tyrosine hydroxylase (tyrosine hydroxylase,TH) in the substantia nigra was measured by Western blot method. Results: pIRESneo-EGFP-BDNF was successfully constructed by double enzyme digestion. The rotation times of MSCs group and BDNF group were significantly decreased than that of PD group (P0.05), especially in BDNF group compared with MSCs group (P0.05). The content of BDNF,TH protein in substantia nigra was significantly increased in BDNF group than in MSCs group after 8 weeks of transplantation (P0.05). Conclusion: pIRESneo-EGFP-BDNF modified bone marrow MSCs can significantly improve the behavioral ability of PD rats and increase the content of BDNF,TH protein in the substantia nigra.
【作者单位】：河北大学生物技术研究中心;河北大学附属医院神经内科;
【基金】：河北省科学技术研究与发展计划资助项目(编号:08206120D)
【分类号】：R-332

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