神经生长因子对人胚胎视网膜色素上皮细胞生长及其DNA合成的影响

The effects of NGF on proliferation and DNA synthesis in cultured human fetal RPE cells

目的 探讨神经生长因子 (NGF)对体外培养的人胚胎视网膜色素上皮 (RPE)细胞的促增殖作用及DNA合成的影响。方法 分别用MTT法测定加入NGF 10 0、2 0 0、30 0 μg/L 48h后人RPE细胞数量的改变和核酸蛋白分析仪测定加入NGF 5 0、10 0、2 0 0、30 0、40 0 μg/L 48h后人RPE细胞DNA质量浓度的变化。 结果 10 0、2 0 0、30 0 μg/LNGF作用 48h后其细胞增殖的A值分别为 ( 0 2 137± 0 0 2 33)、( 0 2 188± 0 0 181)、( 0 2 32 2± 0 0 16 4) ,与对照组 ( 0 1897± 0 0 15 2 )相比 ,差异有显著性 (q test ,P <0 0 5 ) ;10 0、2 0 0、30 0、40 0 μg/LNGF作用 48h后其细胞DNA质量浓度分别为 ( 981 2 2 0 4± 12 3 5 35 7)、( 1375 8484± 2 44 4 718)、( 16 5 8 70 71± 176 9381)、( 2 35 3 0 86 3± 6 0 9 90 6 4) μg/ml ,与对照组 ( 6 6 6 8188± 14 1 330 2 ) μg/ml相比差异有显著性 (q test,P <0 0 5 )。结论 NGF可促进人RPE细胞增殖及促进细胞DNA合成。

ObjectiveTo investigate the effects of nerve growth factor (NGF) on proliferation and DNA synthesis in cultured human fetal retinal pigment epithelium (hRPE) cells in vitro.MethodsPrimary culture and subculture of hRPE cells were established in vitro firstly.Cultured hRPE cells were treated with NGF by various concentrations 50,100,200,300,400 μg/L (final concentration) for 48 h.After 48 h,cells proliferation were measured with methyl thiazolyl tetrazolium (MTT) assay method and the amount of DNA was determined by the absorbance at 280 nm of nucleic acid and protein analysis.ResultsThe A values of 100,200,300 μg/L of NGF were ( 0.213 7±0.023 3),( 0.218 8±0.018 1),( 0.232 2± 0.016 4),q value was 3.63,4.40,6.42 and P values were 0.015,0.000,0.000 (q-test) as compared to ( 0.189 7± 0.015 2) of A value of 0 μg/L NGF respectively.The DNA concentrations for 100,200,300,400 μg/L NGF were ( 981.220 4± 123.535 7),( 1 375.848 4± 244.471 8),( 1 658.707 1± 176.938 1),( 2 353.086 3± 609.906 4)μg/ml,q values were 3.63,8.20,11.47,19.46,P values were 0.024,0.000,0.000,0.000 (q-test) as compared to ( 666.818 8± 141.330 2) μg/ml of DNA concentration for 0 μg/L NGF respectively.ConclusionNGF could stimulate the proliferation and DNA synthesis in cultured hRPE cells in vitro in a dose-dependent manner.