应用流式细胞术检测17β-雌二醇对H_2O_2诱导的视网膜神经细胞凋亡的保护作用

Protective effect of 17β-estrodiol on retina neuron cell apoptosis induced by H_2O_2 by the method of FACS

目的用流式细胞技术(FACS)检测H2O2诱导的视网膜神经细胞凋亡以及17β-雌二醇(βE2)对细胞的保护作用。方法取新生24 h内的SD大鼠进行视网膜神经细胞的原代培养,待细胞培养4-5 d,经不同因素处理后,用MTT法检测细胞存活率,用FACS法检测细胞凋亡率。结果200μmol/L的H2O2能有效诱导视网膜神经细胞凋亡;低浓度的H2O2不能诱导细胞凋亡,但高浓度的H2O2诱导细胞凋亡的同时也导致了正常细胞比率的锐减;10μmol/L的βE2能有效对抗H2O2诱导的视网膜神经细胞凋亡;低浓度的βE2对细胞未表现出保护作用,高浓度的βE2对细胞反而表现出毒性作用。结论一定浓度的βE2在一定范围内能对抗H2O2诱导的视网膜神经细胞凋亡,表现对神经细胞保护的作用。

Objective To observe H2O2-induced neuronal cell apoptosis and neuroprotection of 17 βE2 by the technique of FACS. Methods The primary retina neuronal cells of neonatal SD rats which were born in 24 h were cultured. After 4- 5 days, we treated the cells using different factors, then observed the cell viability by MTT method and the cell apoptosis by FACS method, respectively. Results H2O2 treatment （200 μmol/L） could effectively induce the cultured primary retina neuron cell apoptosis. The low concentration of H2O2 could not affect the cells, but the high concentration treatment could induce cell apoptosis, at the same time leading to normal cell decrease sharply;βE2treatment （10 μmol/L） showed neuroprotection on the primary cultured retina neuron cells, but the low concentration of βE2 had no effects, and the high concentration of βE2 manifested toxic action on the cells. Conclusion The limited concentration of βE2 shows effects of anti-apoptosis and neuroprotection on the cultured primary retina cells.