摘要
目的 探讨用N-甲基-D-门冬氨酸(N-methyl-D-aspartate,NMDA)诱导体外目的 探讨用 N-甲基 -D-门冬氨酸 (N-methyl-D-aspartate,NMDA)诱导体外培养的人胚胎视网膜色素上皮 (retinal pigment epithelium,RPE)细胞的凋亡。方法 加入终浓度分别为 1 0、50、1 0 0、2 0 0、3 0 0 μmol· L-1的 NMDA作用 72 h;3 0 0 μmol· L-1NM-DA作用 1 2、2 4、3 6、48、72 h后用透射电镜定性观察凋亡细胞 ,并用吖啶橙荧光染色进行定量和定性分析。结果 经 2 0 0、3 0 0 μmol· L-1NMDA作用 72 h后 RPE细胞凋亡指数分别为 (51 .0 0 0± 1 .4491 ) %和 (74.0 0 0± 1 .71 2 7) %与对照组 (2 .0 0 0 0± 0 .42 1 6) %相比 ,差异有显著性 (t=1 0 .0 66、1 4.790 ,P=0 .0 0 0、0 .0 0 0 ) ,随着 NMDA浓度的增加 ,RPE细胞凋亡数逐渐增多 ;3 0 0 μmol·L-1NMDA作用 2 4、3 6、48、72 h后 RPE细胞凋亡指数分别为(9.0 0 0 0± 0 .73 79) %、 (1 3 .0 0 0 0± 1 .0 593 ) %、(2 5.0 0 0 0± 0 .70 71 ) %、(53 .0 0 0 0±0 .82 3 3 ) %与对照组 (2 .0 0 0 0± 0 .42 1 6) %相比差异有显著性 (t=2 .0 1 4、3 .1 65、6.61 8、1 4.676,P=0 .0 50、0 .0 0 3、0 .0 0 0、0 .0 0 0 )。结论 NMDA能诱导体外培养的人胚胎
<Abstrcat>ObjectiveTo investigate the involvement of apoptosis in NmethylDaspartate(NMDA) induced excitotoxicity in cultured human fetal retinal pigment epithelium(RPE) cells in vitro. MethodsPrimary culture and subculture of human fetal retinal pigment epithelium cells were established in vitro first.Then the cultured RPE cells were treated with NmethylDaspartate(NMDA) in various concentrations 10μmol·L-1, 50μmol·L-1, 100μmol·L-1, 200μmol·L-1, 300μmol·L-1 (final concentration) for 72h and 300μmol·L-1 NMDA for 12?24?36?48?72h , the quantitative and the qualitative analysis of apoptotic RPE cells were assessed using an acridine orange (AO) staining method and transmission electron microscopy (TEM) respectively. ResultsAO staining in apoptotic RPE cells showed typical apoptosis with chromation condensation and reduction of nuclear size and cell volume and orangestained condensed nucle. The RPE apoptotic numbers of 200?300μmol·L-1 NMDA were (51.0000±1.4991)%, (74.0000±17127)% compared to (2.0000±04216)% of number of 0μmol·L-1 NMDA respectively, t value was 10.066?14.790 and P value was 0.000?0.000 (ttest) after 72h; The RPE apoptotic numbers of 24?36?48?72h was (9.0000±0.7379)%? (130000±10593)%?(25.0000±0.7071)%?(53.0000±0.8233) % compared to (2.0000±04216)% of number of 12h respectively, t value was 2.014?3.165?6.618?14676 and P value was 0.050?0.003?0.000?0.000(ttest) by 300μmol·L-1 NMDA , the more concentration of NMDA, the more apoptosis of RPE cells. ConclusionThe data suggested that NMDA could induce apoptosis in cultured RPE cells in vitro in a dose and coursedependent manner.
出处
《眼科新进展》
CAS
2002年第5期309-311,共3页
Recent Advances in Ophthalmology
