氨基甲酰促红细胞生成素对氧糖剥夺条件下神经干细胞保护作用的研究

Protective effect of carbamylated erythropoietin on neural stem cells in oxygen-glucose deprivation conditions

目的 研究氨基甲酰促红细胞生成素（CEPO）对体外神经干细胞缺氧缺血性损伤的保护作用.方法 从孕14-16 d大鼠获得神经干细胞,经过培养并传代,对所获细胞的自我增殖、自我更新及多分化潜能进行检测.取第3代神经干细胞中添加CEPO,在氧糖剥夺（OGD）条件下培养2 h.通过TUNEL法计数神经干细胞凋亡率和MTT法检测神经干细胞的存活情况.采用酶联免疫吸附试验（ELISA）法检测神经干细胞分泌IFN-γ的情况.应用流式细胞仪检测MHC-Ⅱ在神经干细胞中的表达.结果 在OGD环境中,细胞凋亡率达58.97%,存活率为39.46%,IFN-γ和MHC-Ⅱ类分子的表达分别为78.47 pg/ml和35.68%,加入CEPO后神经干细胞的凋亡率下降至30.15%,存活率升高至75.84%,IFN-γ（15.35 pg/ml）和MHC-Ⅱ类分子的表达均下降（9.77%）.结论 CEPO对神经干细胞缺氧缺血性损伤具有明显的保护作用,而且对炎症分子IFN-γ和免疫分子MHC-Ⅱ类分子的表达具有调节作用.

Objective To observe the protection of carbamylated erythropoietin （CEPO） on neural stem cells （NSCs） impaired by oxygen glucose deprivation （OGD）. Methods The NSCs of 14-16 days were isolated, cultured, and passaged. Proliferation, self-renewal ability and multipotency of differentiation of NSCs were examined. CEPO was added to the third passage of NSCs culture exposed to OGD condition for 2h. The apoptosis of NSCs was studied by TUNEL. The proliferation of NSCs was evaluated by MTT method. The level of IFN--γ was detected by ELJSA. Flow cytometry was applied to measure the expression of MHC-Ⅱ in NSCs. Results Under OGD condition, apoptosis of NSCs was 58.97% , and survival rate was 39.46% , and expressions of IFN--γ and MHC- Ⅱ were 78.47 pg/ml and 35.68% , respectively. Supplementation of CEPO displayed significant decrease of apoptosis （30. 15% ） and increase of survival rate （75. 84% ）. The level of IFN-7 （15.35 pg/ml） and expression of MHC-H （9.77%） were decreased significantly in CEPO-treated group. Conclusion CEPO can significantly protect NSCs against hypoxia and ischemia, and meanwhile, CEPO can regulate the expression of IFN--γ and MHC- Ⅱ.