目的观察大鼠酪氨酸羟化酶(tyrosinehydroxylase,TH)修饰的骨髓基质源性神经元样干细胞(neuronoid stem cells derived from bone marrow stem cells,NdSCs-D—BMSCs)在脑室移植途径下对帕金森病(Parkinson disease,PD)大鼠的治疗作用。方法将酶切鉴定后的新构建质粒pEGFP—C2-TH经电穿孔法转染培养第8天NdSCs—D—BMSCs,注射到PD大鼠模型右侧脑室,观察大鼠行为学变化,移植细胞在大鼠脑组织内的迁移,以及高效液相方法检测脑内DA含量。结果质粒pEGFP—C2-TH转染NdSCs—D—BMSCs移植后10周,PD大鼠症状显著改善,DA恢复至正常水平33.0%,移植细胞可以在PD大鼠脑内存活,并出现远处迁移。结论TH修饰的大鼠NdSCs—D—BMSCs经脑室移植对PD大鼠具有明显的治疗作用,为临床中腰椎穿刺干细胞移植的应用提供实验依据。
Objective To observe the therapeutic effects of tyrosine hydroxylase (TH) -transfected neuronoid stem cells derived from bone marrow stem cells ( NdSCs - D - BMSCs) on Parkinson's disease (PD) by microinjection into the cerebral ventricles (CV). Methods After identification by enzyme digestion, the constructed plasmid pEGFP - C2 - TH was transfected into 8 - day - cultured NdSCs - D - BMSCs by electroporation resulting in the coexpression of green fluorescent protein (GFP) and TH. The TH - transfected cells were injected into the right CV of PD rats. The changes in locomotor activity of PD rats and the migration of transplanted cells in cerebral tissue were monitored and cerebral DA levels were assayed by high performance liquid chromatography (HPLC). Results Ten weeks after transplantation, the symptoms of PD rats were significantly improved and DA levels were restored 33% of control. The transferred ceils showed excellent survival rates in PD rat brains and distant migration was observed. Conclusion CV transplantation of TH - transfected NDSCs - D - BMSCs has obvious therapeutic effects on PD rats. This study could provide evidence for future transplantation route of stem cell transplantation through lumbar puncture.