大鼠胚肾组织促帕金森病移植神经元存活的实验研究

Fetal kidney tissue increases the survival of grafted dopamine neurons in Parkinsonian rats

目的 :探讨大鼠胚肾组织对帕金森病移植神经元的促存活作用。方法 :用神经毒剂 6 OHDA单侧损毁SD大鼠左侧中脑被盖腹侧区 (VTA)和黑质致密部 (SNC)建立动物模型。实验分 3组 :A组将胚鼠腹侧中脑组织 (VM )预置于胚肾组织培养液中 ;B组将VM组织预置于胶质细胞源性神经营养因子 (GDNF)营养液中 ;C组将VM组织预置于DMEM营养液中。 6h后移植 ,于移植后 1、4、7d将动物处死 ,采用原位末端标记法 (TUNEL)结合流式细胞术及免疫组织化学方法对 3组移植神经元凋亡和存活情况进行定量分析。结果 :根据TUNEL、酪氨酸羟化酶 (TH)染色和流式细胞仪检测结果 ,A、B移植组较C移植组移植神经元凋亡率降低 ,凋亡细胞数减少 ,纹状体移植针道和针道周围都可见大量TH阳性细胞 ,而且TH阳性神经元数量明显增多。另外 ,A移植组和B移植组TH阳性神经元和凋亡细胞数比较也存在统计学差异。结论 :胚肾组织分泌物对帕金森病移植神经元具有促存活作用 ;胚肾组织培养液较单一生长因子作用好。

Objective To investigate whether fetal kidney tissue is beneficial for the survival in intrastriatum transplants of fetal ventral mesencephalic cell suspension.Methods Nigral dopaminergic neurons was lesioned in adult sprague dawley rats by injecting 6 hydroxydopamine into nigra.Four weeks following the procedure,36 animals were selected and divided into three groups:group A(in kidney tissue culture n =12);group B (in GDNF culture n =12);group C (in DMEM culture n =12);1,4,7 days after grafting,the rats were sacrificed for TUNEL,TH immunohischemistry staining and FACM detection.Results Fetal kidney tissue pretreatment increased the survival of dopamine neuron,growth factors deprived from kidney tissue reduced the apoptotic rate of transplanted cells.TH immunohistochemistry revealed THir positive neurons were also significantly more in A,B compared to C group ( P < 0.05 ).Conclusion Growth factors deprived from kidney tissue reduce the apoptotic rate of transplanted cells and increase the survival of dopamine neurons.