神经生长因子对过氧化氢诱导人胚胎视网膜色素上皮细胞凋亡的保护作用

Nerve growth factor prevents apoptosis induced by hydrogen peroxide in cultured human fetal retinal pigment epithelial cells

目的 探讨神经生长因子 (nerve growth factor,NGF)对体外培养的人胚胎视网膜色素上皮 (human fetal retinal pigm ent epithelium,h FRPE)细胞凋亡的保护作用。方法用传代培养的 h FRPE细胞 ,30 0、5 0 0 μmol· L- 1 过氧化氢 (hydrogen peroxide,H2 O2 )建立诱导的 h FRPE细胞凋亡模型 ,并用吖啶橙 (acridine orange,AO)荧光染色计数和透射电镜 (transmission electron microscope,TEM)观察 NGF对凋亡细胞的保护作用。结果 用AO荧光染色及 TEM均观察到经 30 0、5 0 0μmol· L- 1 H2 O2 处理 6 5 h后的 h FRPE细胞出现典型的凋亡形态学改变。 30 0μm ol· L- 1 H2 O2 作用 6 5 h后 h FRPE细胞凋亡指数为4 5 .6 2 %± 1.4 1%和 30 0μmol· L- 1 H2 O2 +40 0μg· L- 1 NGF作用 6 5 h后的 h FRPE细胞凋亡指数 2 2 .79%± 1.30 %相比 ,差异有显著性 (q=4 .84 4 5 ,P=0 .0 0 0 0 ) ;5 0 0μmol· L- 1H2 O2 作用 6 5 h后 h FRPE细胞凋亡指数为 6 6 .2 2 %± 1.86 %和 5 0 0 μm ol· L- 1 H2 O2 +40 0 μg· L- 1 NGF作用 6 5 h后的 h FRPE细胞凋亡指数 4 2 .5 6 %± 1.2 1%相比差异有非常显著性的意义 (q=5 .114 2 ,P=0 .0 0 0 0 )。结论 NGF能拮抗 H2 O2 诱导的 h

Objective To investigate the protective effect of nerve growth factor (NGF) on apoptosis of cultured human fetal retinal pigment epithelial (hFRPE) cells induced by hydrogen peroxide (H 2O 2) in vitro. Methods Subcultured hFRPE cells were treated with 300?500μmol·L -1 of H 2O 2 to be established cell apoptotic model. The protective effect of NGF on apoptosis of cultured hFRPE cells were assessed by an acridine orange(AO) staining method and transmission electron microscopy(TEM).Results H 2O 2 300?500μmol·L -1 exposured to hFRPE cells for 65h elicited typical apoptosis morphological changes including condensed chromation, nuclear fragmentation and reduction of nuclear size and cell volume. There was a significant difference in hFRPE cells with apoptosis between 300μmol·L -1 H 2O 2 45.62%±1.41% and 300μmol·L -1 H 2O 2 + 400μg·L -1 NGF 22.79%±1.30% groups after 65h ( q =4.844 5, P =0 000 0);There was a significant difference in hFRPE cells with apoptosis between 500μmol·L -1 H 2O 2 66.22%±1.86% and 500μmol·L -1 H 2O 2 + 400μg·L -1 NGF 42.56%±1.21% groups after 65h as well ( q =5.114 2, P =0.000 0). There had been a decrease in the number of apoptotic hFRPE cells and some normal hFRPE cells were seen among the apoptotic hFRPE cells and morphology and ultrastructure of hFRPE cells returned to near normal after by treatment with mentioned different concentration of NGF at different time.Conclusion NGF has an protective effect on H 2O 2 induced hFRPE cells apoptosis.