重组人促红细胞生成素预处理对黑质多巴胺能神经元凋亡的影响

目的研究重组人促红细胞生成素(rhEPO)对离体帕金森病模型中黑质多巴胺神经元凋亡的影响。方法以6-羟基多巴胺(6-OHDA)为毁损剂建立大鼠离体帕金森病(PD)模型。用6u/mlrhEPO预处理黑质多巴胺神经元,然后用免疫组化方法观察黑质中酪氨酸羟化酶(TH)免疫反应阳性细胞数和半胱天冬酶-3(Caspase-3)免疫反应阳性细胞数的变化,TUNEL法观察黑质中多巴胺神经元的凋亡情况。结果与6-OHDA组(44.2±5.0)相比,rhEPO预处理组TH免疫反应阳性细胞(63.8±6.2,P<0.01)增多;与6-OHDA组(22.3±2.8)相比,rhEPO预处理组多巴胺神经元中Caspase-3表达减少,Caspase-3免疫反应阳性细胞染色较淡,数量减少(13.7±1.8,P<0.01);与6-OHDA组(20.3±3.1)相比,rhEPO预处理组TUNEL阳性细胞染色较淡,数量减少(10.7±1.5,P<0.01)。结论rhEPO预处理可以减轻6-OHDA对离体帕金森病模型中多巴胺神经元的损伤,其机制可能与rhEPO抑制黑质多巴胺神经元凋亡有关。

Effect of preconditioning with recombinant human erythropoietin on dopaminergic neuronal apoptosis in substantia nigra of rat with Parkinson 's disease induced by 6-hydroxydopamine in vitro

Objective To study the effect of preconditioning with recombinant human erythropoietin （rhEPO） on dopaminergic neuronal apoptosis in substantia nigra （SN） of Parkinson＇s disease （PD） induced by 6-hydroxydopamine（6-OHDA） in vitro. Methods PD models in vitro were established by incubating rat midbrain slices containing SN in artificial cerebrospinal fluid II（ACSFII） containing 6-OHDA （0.05mM） . After preconditioning with rhEPO at concentration of 6u/ml for 6 hours , the immunohistochemical technique was used to detect the expression of tyronsine hydroxylase （TH） and Caspase-3 in SN. Meanwhile, TUNEL method was used to detect dopaminergic neuronal apoptosis. Results rhEPO caused an increase of TH-IR positive cells from 44.2 ± 5.0 in the 6-OHDA group to 63.8 ±6.2 in the 6-OHDA ＋ rhEPO group（ P 〈 0.01 ）. The expression of Caspase-3 in dopaminergic neurons in SN in the 6-OHDA ＋ rhEPO group decreased, Caspase-3-IR positive cells in the 6-OHDA ＋ rhEPO group were less lightly stained than those in the 6-OHDA group ;and rhEPO caused a decrease of Caspase-3-IR positive cells from 22.3 ±2.8 in the 6-OHDA group to 13.7 ±1.8 in the 6-OHDA ＋ rhEPO group （ P 〈0.01 ）. TUNEL positive cells in the 6-OHDA ＋ rhEPO group were less lightly stained when compared with those in the 6-OHDA group;and rhEPO caused a decrease of TUNEL positive cells from 20.3± 3.1 in the 6-OHDA group to 10.7 ± 1.5 in the 6-OHDA ＋ rhEPO group （ P 〈 0.01 ）. Conclusion Preconditioning with rhEPO can ameliorate the damage which is produced by 6-OHDA to DA neurons in SN of PD models in vitro,it seems likely that the protection of rhEPO is associated with inhibition of DA neuronal apoptosis in SN.