苏拉明联合地塞米松对体外培养的视网膜色素上皮细胞增殖的影响

Effects of suramin combined with dexamethasone on proliferation of cultured retinal pigment epithelial cells in vitro

目的:探讨苏拉明(suramin,Sur)联合地塞米松(dexamethasone,Dex)对体外培养的猪视网膜色素上皮(retinal pigment epithelium,RPE)细胞增殖的影响。方法:选择0.05,0.1,0.2,0.4g/L Sur单独作用及联合0.2g/LDex作用于RPE细胞4d,MTT比色法检测对RPE细胞增殖的影响;0.2g/L Sur和0.2g/L Dex分别作用于RPE细胞,流式细胞术检测细胞周期变化;Sur浓度为0.2g/L,0.4g/L及0.2g/L Sur联合0.2g/L Dex时,透射电镜观察细胞超微结构情况。结果:MTT比色法检测表明各浓度Sur对RPE细胞有抑制作用(P<0.05),呈剂量效应关系;0.2g/L Dex与各浓度Sur联合应用对RPE细胞的抑制率由单独应用Sur的16.0%,27.3%,40.5%,64.1%增至42.3%,52.1%,65.6%,78.8%。流式细胞检测提示Sur将细胞阻滞于G2/M期(P<0.01);Dex将细胞阻滞于S期(P<0.01)。透射电镜结果显示,Sur浓度为0.2g/L时,单独应用及联合0.2g/L Dex应用,RPE细胞结构无明显变化,而Sur0.4g/L时,细胞表面微绒毛减少,细胞核皱缩。结论:Sur联合Dex能有效抑制体外培养的RPE细胞增殖。

AIM： To investigate the effects of suramin combined with dexamethasone on proliferation of cultured pig retinal pigment epithelial（RPE） cells. METHODS： Cultured RPE cells were treated with different concentrations of suramin （0.05,0.1,0.2,0.4g/L） lonely and combined with 0. 2g/L dexamethasone for 4 days. The inhibition ratio of RPE cells was measured by tetrazolium （MTT） colorimetric assay. 0. 2g/L suramin and 0. 2g/L dexamethasone were added to RPE cells respectively, flow cytometry（FCM） analysis was used to examine RPE cells cycles. The ultrastructure of cells treated whit 0.2g/L suramin, 0.2g/L suramin combined with 0. 2g/L dexamethasone and 0. 4g/L suramin were observed by transmission electronic microscopy（TEM）. RESULTS： Suramin could inhibit the proliferation of RPE cells in a dose-dependent manner. The inhibition ratios of RPE cells treated with suramin combined with 0. 2g/L dexamethasone （42. 3%, 52. 1%, 65. 6%, 78. 8%） were much higher than those treated with suramin （16.0%, 27.3% ,40.5%, 64.1% ）. FCM revealed that the cells were blocked in G2/M phase in their cell cycle by suramin（P〈 0.01 ） and blocked in G2/M phase by dexamethasone（ P〈 0. 01 ）. The ultrastructure of cells were well at the concentrations of 0. 2g/L suramin and 0. 2g/L suramin combined with 0. 2g/L dexamethasone, but at theconcentration of 0.4g/L, significant reduction of microvilli and crenulation of cell nucleus were observed. CONCLUSION- Suramin combined with dexamethasone can effectively inhibit RPE cells growth in vitro.