中文摘要：

目的在模拟人中脑黑质的多种原代细胞培养体系中，研究铁对多巴胺能神经元的损伤及其神经免疫炎症机制。方法采用5、25和100μmol／L的FeCl2（Fe^2＋）：（1）处理中脑原代神经元-小胶质细胞-星形胶质细胞混合培养体系，7d后计数酪氨酸羟化酶（TH）（＋）多巴胺能神经元及抗神经特异性核蛋白抗体（Neu—N）（＋）的全部神经元的数量，观察神经元形态，研究Fe^2＋对多巴胺能神经元的选择性损伤；（2）同时处理中脑原代神经元-小胶质细胞-星形胶质细胞混合培养体系和神经元-星形胶质细胞培养体系，7d后计数TH（＋）多巴胺能神经元的数量，研究小胶质细胞在Fe^2＋选择性损伤多巴胺能神经元中的作用；（3）处理原代小胶质细胞培养体系，测量细胞外超氧化物（O2^-）及细胞内活性氧类物质（iROS），研究Fe^2＋激活小胶质细胞导致的功能变化；（4）处理中脑原代神经元-小胶质细胞．星形胶质细胞混合培养体系，7d后计数抗补体受体-3（OX-42）（＋）小胶质细胞的数量，观察细胞形态，研究Fe^2＋激活小胶质细胞导致的形态学变化。结果（1）5、25和100~mol／LFe^2＋处理组TH（＋）多巴胺能神经元的数量分别为对照组的89％、70％和55％，其中25、100μmol／LFe^＋处理组与对照组比较差异具有统计学意义（F=12．047，P〈0．01）；多巴胺能神经元胞体皱缩，胞质淡染，神经突起数目减少；（2）5、25和100μmol／LFe^2＋处理组Neu—N（＋）全部神经元的数量分别为对照组的100％、104％和101％，与对照组比较差异均无统计学意义；5、25和100μmol／LFe^2＋处理组Neu—N（＋）全部神经元的数量与TH（＋）多巴胺能神经元数量的差值分别为11％、34％和46％，其中25和100μmol／LFe^2＋处理组两种神经元数量的差异具有统计学意义（分别为t=8．098，P〈0．05；t=11．218，

英文摘要：

Objective To investigate the role and neuroinflammatory mechanism of iron on dopamine ( DA) neurons in multiple primary midbrain cultures that mimic human substantia nigra pars compacta.Methods Ferrous chloride ( Fe2+ ) with the desired concentrations of 5,25 and 100 μmol/L was used to ( 1 ) treat primary midbrain neuron-microglia-astroglia cultures for 7 days and the numbers of DA neurons and total neurons were counted after tyrosine hydroxylase (TH) and neuron-specific neuclear protein neurons in 5,25 and 100 μmol/L Fe2 + -treated groups were 89%,70% and 55% of control group,and 25,100 μmol/L Fe2+ significantly decreased DA neuronal numbers compared with control group ( F = 12.047,P ＜0.01);DA neuronal bodies were shrunk and smaller,cytoplasmic stainings were reduced,neuronal dendrites were decreased;(2) The numbers of Neu-N ( + ) total neurons in 5,25 and 100 μmol/L Fe2+-treated groups were 100%,104% and 101% of control group and Fe2+ did not decrease DA neuronal numbers compared with control group (t =4.458,P ＞ 0.05 );5,25 and 100 μmol/L Fe2+-induced difference between total neurons and DA neurons were 11%,34% and 46%,and 25 and 100 (Amol/L Fe2+ produced significant difference(t =8.098,P ＜0.05;t = 11.218,P＜0.05);(3) In primary midbrain neuron-microglia-astroglia and neuron-astroglia cultures,the numbers of DA neurons in 25 μmol/L Fe+-treated group were 70% and 98% of control group,respectively.The difference between two groups was 28%,which was statistically significant (t =8.061,P＜0.05);The numbers of DA neurons in 100 μmol/L groups were 183%,190 % and 240% of control group,and 25 and 100 μmol/L Fe2 + significantly increased microglial numbers compared with control group ( F = 6.101,P ＜ 0.01 );dramatic changes of microglial morphology were indicated by the enlarged cell bodies and irregular shape.Conclusions Fe2 + provokes selective DA neuronal damage and microglia are the mediators of the neurotoxic effect,which may be due to microglial over-activation featured by the significant producti