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目的:观察移植pIRESneo-EGFP-BDNF修饰骨髓间充质干细胞(MSCs)对帕金森病模型大鼠TH表达的影响。方法:采用电穿孔法将pIRESneo-EGFP-BDNF转染至骨髓MSCs;通过脑内注射6-OHDA制备PD大鼠模型,随机分为Sham组,PD组,MSCs组,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,移植BDNF修饰骨髓MSCs在大鼠脑组织内的迁移,采用免疫组化及Western blot方法测定各组大鼠中脑黑质TH蛋白表达的变化。结果: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(P<0.05),以BDNF组较MSCs组比较改善更为明显(P<0.05),移植细胞干预PD模型大鼠8周后中脑黑质TH表达,BDNF组较MSCs组、PD组明显增高(P<0.05),移植pIRESneo-EGFP-BDNF修饰骨髓MSCs可以在PD大鼠脑内存活,并迁移至远处。结论:经侧脑室移植pIRESneo-EGFP-BDNF修饰的骨髓MSCs通过提高中脑黑质TH的活性,能够明显改善PD模型大鼠的行为能力。
Objective: To observe the effect of transplanted pIRESneo-EGFP-BDNF modified bone marrow mesenchymal stem cells (MSCs) on TH expression in Parkinson’s disease model rats. METHODS: The pIRESneo-EGFP-BDNF was transfected into bone marrow mesenchymal stem cells by electroporation. PD rats were injected with 6-OHDA through intracerebral injection. The rats were randomly divided into Sham group, PD group, MSCs group and BDNF group. MSCs or pIRESneo-EGFP-BDNF were used to modify bone marrow MSCs. Apomorphine was injected intraperitoneally at 2, 4, and 8 weeks after the operation to induce PD rats to rotate. BDNF-modified bone marrow MSCs were transplanted into the brain of rats. Immunohistochemistry Western blot was used to determine the changes of TH protein expression in substantia nigra of rats in each group. Results: Compared with PD group, the numbers of rotations of MSCs group and BDNF group were significantly decreased (P <0.05) at 2, 4, 8 weeks after transplantation, and more significantly in BDNF group than those in MSCs group (P <0.05) After 8-week intervention, the expression of TH in substantia nigra was increased in BDNF-treated group compared with MSCs group and PD group (P <0.05). The transplantation of pIRESneo-EGFP-BDNF modified bone marrow MSCs could survive and migrate in the brain of PD rats To the distance. CONCLUSION: MSCs modified with pIRESneo-EGFP-BDNF can significantly improve the behavioral ability of PD model rats by increasing TH activity in midbrain substantia nigra.