摘要
目的:探讨大鼠酪氨酸羟化酶(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)分别在脑室移植途径及纹状体移植途径下对帕金森病(Parkinson's 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8d NdSCs-D-BMSCs后,观察绿色荧光蛋白及TH的共表达,而后分别注射到PD大鼠模型右侧纹状体及右侧脑室,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的含量。结果:质粒pEGFP-C2-TH转染NdSCs-D-BMSCs后第5d,绿色荧光蛋白表达59.0%,与TH共表达率为83.5%;移植后10周,纹状体移植组及脑室移植组大鼠症状均显著改善,DA含量较模型对照组显著提高(P<0.01),分别恢复至正常水平46.6%、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在纹状体移植及脑室移植途径下均对PD大鼠具有明显的治疗作用,为临床中多种移植途径的选择,特别是经脑室穿刺干细胞移植提供实验依据。
Objective ：To investigate the therapeutic effects of tyrosine hydroxylase（TH） transfected neu-ronal stem cells derived from bone marrow stem ceils （NdSCs - D - BMSCs） on Parkinson＇s disease （PD） through different transplantation protocols, including microinjection into the cerebral ventricles （CV） and the striatum （ST）. Methods After identification by enzyme digestion, the constructed plasmid pEGFPC2 -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 either the right ST or 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 Five days after plasmid pEGFP - C2 - TH transfection into NdSCs - D - BMSCs, GFP was expressed in 39.0% of the cells and the rate oi co - expression with TH was 85.3%. len weeks following transplantation, the symptoms of PD rats in both groups were significantly improved and DA levels were re- stored to 46.60/0 in：ST group and 33% in CV group of control. The transferred cells showed exceUent survival rates in PD rat brains and distant migration was observed. Conclusion Both CV and ST transplantation of TH - transfected NDSCs - D - BMSCs has obvious therapeutic effects on PD rats. This study could provide evidence for CV transplantation route selection.
出处
《新疆医学》
2012年第6期21-27,共7页
Xinjiang Medical Journal
关键词
基因治疗
骨髓基质源性神经元样干细胞
帕金森病
大鼠
移植途径
Gene therapy
Neuronal stem cells derived from bone marrow stem cells
Parkinsong dis-ease
Rats
Transl31antation route
