携带GAD65的永生化神经前体细胞移植治疗癫痫大鼠

Transplantation of genetically engineered cell lines into hippocampus in rat epileptic model induced by pflocarpine

目的将构建好的携带GAD65的永生化神经前体细胞（INPCs—GAD65）移植到癫痫大鼠的脑内，观察这些细胞在癫痫大鼠脑内的存活及对癫痫大鼠的治疗作用。方法INPCs—GAD65用超顺磁性氧化铁纳米粒子（SPIO）标记后，MRI扫描追踪观察细胞的存活状况；观察对照组、实验组大鼠在行为学、脑电图的改变；免疫组织化学观察其分化。结果行1．5TMRI检查，见移植处在T2WI呈低信号改变，移植后7周仍可见移植处的低信号改变，随时间延长信号变浅；与对照组比较，行为学观察发现自发痫性发作次数减少差异有统计学意义；脑电图改善，痫波发放减少；移植INPCs-GAD65可分化为神经元和胶质细胞。结论在癫痫大鼠脑内移植合成和分泌GABA的永生化神经前体细胞能在脑内存活及分化，并能改善痫性发作。

Objective Transplant genetically engineered cell lines to synthesize and release GA- BA by immortalized neural progenitor cells （INPCs） in rats, can survive and differentiate in the hippocampus in the epileptic rats model induced by pilocarpine, have ability to suppress spontaneous seizures. Methods The INPCs were transfected with the GABA-synthesizing enzyme GAD （65） cDNA （INPCs- GAD65）, Labeled INPCs-GAD65 with superparamagnetic iron oxide （SPIO）, SPIO-labeled cells were tracked with 1.5T MRI scanner in vivo;Behavior were monitored for 6 days after surgery;The change exam- ined with the electroencephalogram recording; The differentiation of transplanted cells was observed with immunohisochemistry. Results Animals that received genetically engineered GABA-producing cells had significantly fewer spontaneous seizures than did that the control animals, Epileptiform spikes was suppressed significantly in the electroencephalogram recording; 1.5 Tesia MRI in vivo displayed that remarkable low signal area on T2WI was seen in the right brain which transplanted with SPIO-labeled cells. SV40Tag-positive cell can change into neurons and astrocytes in the hippocampus. Conclusion These data demonstrate that genetically engineered cells could suppress spontaneous seizures in an accepted model of temporal lobe epilepsy. This is an important step toward defining a clinical potential for this approach in epilepsy.