N-甲基-D-门冬氨酸对体外培养的视网膜色素上皮细胞增殖及DNA合成的影响

EFFECTS OF N-METHYL-D-ASPARTATE ON PROLIFERATION AND DNA SYNTHESIS IN CULTURED HUMAN FETAL RETINAL PIGMENT EPITHELIUM CELLS

目的 :探讨N -甲基 -D -门冬氨酸对体外培养的人胚胎视网膜色素上皮细胞的抑制作用及DNA合成的影响。方法 :分别用MTT法测定加入NMDA 10 ,50 ,10 0 ,2 0 0 ,3 0 0 ,40 0 μmol/L 72h后RPE细胞数量的改变和核酸蛋白分析仪测定加入NMDA 10 0 ,150 ,2 0 0 ,2 50 ,3 0 0 μmol/L 72h后RPE细胞DNA浓度的变化。结果 :50 ,10 0 ,2 0 0 ,3 0 0 ,40 0 μmol/LNMDA作用 72h后其细胞OD与对照组相比 ,差异有显著性 (t检验 ,P <0 .0 5) ;10 0 ,150 ,2 0 0 ,2 50 ,3 0 0 μmol/LNMDA作用 72h后其细胞DNA与对照组相比 ,差异有显著性 (t检验 ,P <0 .0 5)。结论 :NMDA可抑制体外培养的RPE细胞增殖及促进细胞DNA合成。

Objective:To investigate the inhibitive effects of N-methyl-D-aspartate(NMDA) on proliferation and DNA synthesis of cultured human fetal retinal pigment epithelium (hRPE) cells in vitro. Methods:Primary culture and subculture of human fetal retinal pigment epithelium cells were established in vitro first. Cultured hRPE cells were treated with NMDA by various concentrations of 10μmol/L,50μmol/L,100μmol/L,200μmol/L,300μmol/L and 400μmol/L (final concentration) for 72h.After 72h, cell proliferation was measured with methyl thiazolyl tetrazolium (MTT) assay method and 100μmol/L,150μmol/L,200μmol/L,250μmol/L and 300μmol/L (final concentration) for same time. After 72h,the amount of DNA was determined by the absorbance at 280 nm of Nucleic Acids δ Protein Analysis. Results:There was a statistically significant difference in OD values of 10μmol/L, 50μmol/L,100μmol/L,200μmol/L,300μmol/L and 400μmol/L NMDA compared to OD value of 0μg/L NMDA ( t -test, P <0.05), respectively; there was a statistically significnat difference in DNA concentrations of 100μmol/L,150μmol/L,200μmol/L,250μmol/L and 300μmol/L NMDA compared to DNA concentration of 0μg/L NMDA ( t -test, P <0.05), respectively. Conclusions:The data suggested that NMDA could inhibit the proliferation and DNA synthesis in cultured hRPE cells in vitro in a dose-dependent manner.