人羊膜上皮细胞移植治疗帕金森病鼠的研究

目的 观察人羊膜上皮细胞在帕金森病鼠移植后的存活情况,以及它对帕金森病鼠旋转行为的改善作用.方法 采用6-羟多巴胺立体定向纹状体注射制作帕金森病鼠模型;51只大鼠随机分三组:人羊膜上皮细胞移植组、假手术PBS对照组以及空白模型对照组.制模成功后第5周用人特异性抗体Nestin和Vimentin检测人羊膜细胞的存活情况,第10周切片观察黑质部TH阳性神经元的变化情况,高效液相色谱--电化学仪测定纹状体多巴胺(DA),高香草酸(HVA),3,4-二羟基苯乙酸(DOPAC)等浓度以及脑脊液DA的含量.结果 人羊膜上皮细胞帕金森病鼠侧脑室内移植可以存活达10周;移植组大鼠旋转数较对照组明显降低(P≤0.01);黑质部TH阳性神经元数量较对照组升高(P≤0.01),纹状体区DA、HVA和DOPAC含量较PBS对照组明显升高(P＜0.01～0.05),移植组脑脊液DA含量较PBS对照组也显著增加(P＜0.01).结论 人羊膜上皮细胞侧脑室移植可以改善帕金森病鼠的旋转行为,其机制可能与其增加纹状体区多巴胺等递质水平有关.

An effect of transplantation of human amniotic epithelial cells on Parkinson disease model rats

Objective To investigate the survival time of human amniotic epithelial cells(HAECs) after being transplanted into the lateral ventricle of Parkinson disease (PD) model rats,and the effect of HAECs on the circling behavior.MethodsThe Parkinson model was made by using stereotactic microinjection of 6-hydroxydopamine(6-OHDA) into the cerebral striatum.51 PD models were randomly divided into three groups:the HAECs group,the phosphate buffer saline(PBS) group and the normal model group.The HAECs survival expression was investigated in the fifth week by using human specific monoclone antibody nestin and vimentin.After being treated for 10 weeks,the influence of HAECs on the substantia nigra's tyrosine hydroxylase (TH) positive cells of all three groups was collected.Finally,the contents of dopamine (DA)and its metabolite dihydroxyphenylacetic acid(DOPAC) and homovanillic acid(HVA) in the striatum were tested with high performance liquid chromatography-electrochemical detection after being treated for 10 weeks.Results The HAECs can survive for 10 weeks after being transplanted into the lateral ventricle of the PD model rats.The circling behavior of the HAECs group significantly ameliorated than the PBS group and normal model group(P≤0.01).The TH immunohistocbemistry showed that the TH positive cells of HAECs group increased than PBS group and normal model group(P≤0.01).The content of DA,HVA and DOPAC of HAECs group increased than PBS group(P＜0.05～0.01).In addition,the content of DA in the eerebrospinal fluid of HAECs group increased than PBS group(P≤0.01).Conclusions The human amniotic epithelial cells transplantation can improve the cycling behavior of the PD models rats.It may be due to the content of increased DA and its metabolite in the striatum of PD models rats.