NT-3基因修饰的MSCs脑内移植对颞叶癫痫大鼠作用的研究

Effects of Intracerebral Transplantation of NT-3-modified Mesenchymal Stem Cells on Temporal Lobe Epilepsy in Rats

目的观察神经营养素-3(Neurotrophin 3,NT-3)基因修饰的间充质干细胞(mesenchymal stem cells,MSCs)脑内移植对颞叶癫痫(temporal lobe epilepsy,TLE)的作用及其对大鼠海马苔藓纤维发芽(mossy fiber sprouting,MFS)的影响。方法将构建成功的LV-GFP-NT-3转染大鼠骨髓MSCs,检测转染后MSCs内NT-3的表达;建立海人酸(Kainic acid,KA)TLE大鼠模型;将NT-3基因修饰的MSCs移植至海人酸TLE大鼠侧脑室模型后,观察其对TLE大鼠行为学、脑电图的影响,并用Timm组织化学染色法观察海马结构MFS;Western blot检测细胞移植后大鼠海马组织中NT-3的表达。结果 LV-GFP-NT-3转染细胞后,MSCs中能够正确过表达NT-3;细胞移植后NT-3-MSCs组TLE大鼠的发作次数减少,发作级别减低;脑电图结果表明TLE发作痫波减少;NT-3-MSCs组大鼠海马CA3区内分子层及始层未见到或极少见到Timm染色颗粒,Timm染色颗粒较对照组和LV-MSCs组显著减少;Western blot检测结果显示NT-3-MSCs组大鼠海马组织中NT-3含量明显增高。结论 NT-3基因修饰的MSCs脑内移植对颞叶癫痫大鼠有明显的抑制作用。

Objective To examine the expression of NT 3 transfected mesenchymal stem cells（MSCs）in the brain of rats with temporal lobe epilepsy（TLE）and to explore the effect of such expression on TLE. Methods Lentivirus（LV）vectors con taining NT-3 gene were constructed,and they were later transfected into rat bone marrow MSCs. The expression levels of NT 3 gene were detected in MSCs. The TLE model was established in rats induced by kainic acid（KA）. Thereafter, the NT-3-modi fled MSCs were transplanted into the lateral ventricle of TLE models. The effects of NT-3-modified MSCs on the behaviors of TLE rats,and EEG were obserued and the hippocampal mossy fiber sprouting wars measured by Timm histochemical stai ning. Western blotting was used to detect the NT-3 expression. Results Transfection of LV-GFP-NT-3 cells could leael to the over-expression of NT-3 in MSCs. After transplantation with NT-3-MSCs,the number of seizures and attacks were found to be reduced. EEG results showed the epileptic seizure waves were reduced,and Timm histochemical staining showed less of hipp ocampal mossy fiber sprouting. Timm staining particles were significantly reduced in NT-3-modified MSCs-transplanted rats a,s compared with those in the control group and the LV-MSCs group. Western blot analysis showed that in hippocampal tissues NT-3 was significantly increased in NT-3-MSCs group. Conclusion NT-3 gene-modified MSCs transplantation can significantly reduce the seizures in rats with TLE.