慢病毒介导核受体相关因子1基因修饰骨髓间充质干细胞移植治疗帕金森病模型大鼠

目的探讨慢病毒介导核受体相关因子1(Nurrl)基因修饰骨髓间充质干细胞(BMSCs)移植至帕金森病(PD)模型大鼠纹状体后对PD大鼠的治疗作用。方法采用神经毒素6-羟多巴胺(6-OHDA)纹状体注射成功制备PD大鼠模型18只,将携带绿色荧光的慢病毒GV287-Nurr1感染大鼠BMSCs,然后随机移植入6只PD模型大鼠的纹状体内(实验组),设生理盐水组(假移植组)6只及感染空慢病毒GV287的BMSCs组(对照组)6只;术后第1、2、4周观察大鼠的行为改善情况;免疫组织化学染色法检测纹状体及黑质中Nurrl和酪氨酸羟化酶(TH)蛋白的表达变化;RT-PCR法检测黑质Nurrl mRNA和TH mRNA的表达变化。结果慢病毒感染后的BMSCs及上清中均检测到Nurr1蛋白;对照组与实验组大鼠在移植后第1、2、4周的旋转行为较假移植组均有所改善,且实验组比对照组改善更明显;实验组纹状体Nurr1阳性细胞有效存活并沿胼胝体向皮质及对侧脑组织迁移;实验组纹状体及黑质损毁侧Nurrl和TH蛋白及mRNA的表达较假移植组和对照组明显增高,差异均有统计学意义(P0.05)。结论慢病毒介导Nurr1基因修饰大鼠BMSCs移植治疗PD大鼠,能有效地改善PD模型大鼠的行为学症状,增加大鼠脑内纹状体和黑质区Nurr1和TH的表达。

Lentiviral vector mediated Nurrl gene-modified mesenchymal stem cells for the treatment of Parkinson's disease rat model

Objective To investigate the therapeutic effects of bone marrow mesenchymal stem cells (BMSCs) modified by nuclear receptor-related factor 1 (Nurrl) gene transplanted into the striatum of Parkinson’s disease rat model. Methods Eighteen rat models of Parkinson’s disease (PD) were successfully induced by 6-hydroxydopamine (6-OHDA) injection into the right striatum. BMSCs infected by GV287-Nurr1 lentivirus carrying green fluorescence were transplanted into the striatum of six PD rats (the experiment group). Randomly set up six PD rats as saline group (the sham-grafted group) and six PD rats as negative lentivirus GV287 infection of BMSCs group (the control group). After 1, 2 and 4 weeks, the behavioral change of the rats was induced by apomorphine (APO). The expression change of Nurrl and TH in the striatum and substantia nigra were examined by immunohistochemical staining, and the expression change of Nurrl mRNA and TH mRNA in the striatum and substantia nigra were detected by RT-PCR. Results The Nurr1 protein was detected in BMSCs and its supernatant after infected by lentivirus GV287-Nurr1. After transplantation 1, 2 and 4 weeks compared to the sham-grafted group the rat rotational behavior of the control group and the experiment group were improved, and that of the experiment group improved more obvious. The implanted Nurr1-positive cells survived and migrated to the cortex and the contralateral brain tissue along the corpus callosum. Compared with the sham-grafted group and the control group, the expression of Nurrl and TH at protein and mRNA levels of the lesion side of the striatum and substantia nigra after transplantation in experiment group were increased significantly. Conclusion Lentiviral vector mediated Nurrl gene-modified BMSCs can effectively improve the symptoms of PD rats and increase the expression of Nurrl and TH in striatum and substantia nigra of Parkinson’s disease rat model.