苏拉明对体外培养的人眼视网膜色素上皮 细胞增生抑制作用的时效性研究

目的观察苏拉明对体外培养的人眼视网膜色素上皮（RPE）细胞增生抑制作用的时效关系，了解它对RPE细胞的作用方式，进一步阐明其防治增生性玻璃体视网膜病变（PVR）的优势。方法9块96孔细胞培养板，每块取12孔，其中实验组和对照组各6孔。2组各孔内均接种浓度为5×104个/ml的RPE细胞0.1 ml。换液后实验组加250 μg/ml苏拉明，对照组不加。4 d后2组均更换成正常培养液；在生物倒置显微镜下观察RPE细胞生长情况；分别于加药后1、2、4 d及撤药后1、2、3、5、7、9 d随机抽取1块培养板终止培养，采用四甲基偶氮唑盐（MTT）比色法检测同一培养板上两组RPE细胞增生情况。随机化区组 t 检验比较两组吸光度值的差异，计算细胞增生抑制率。结果倒置显微镜下，对照组RPE细胞在接种后第7天完全融合。实验组细胞间隙较对照组稍增大，接种13 d时细胞仍未融合。实验组RPE细胞加入苏拉明后第1天，增生抑制率为14.85%，第4天达最高25.79%；撤药后第1天下降到12.35%，然后逐渐又上升，到撤药后第3~5天达顶峰超过20%，之后逐渐回落，到第9天达14.71%。结论苏拉明对血清诱导的RPE细胞增生具有长时间的抑制作用，特别是在撤除药物后还能再次引起后抑制作用，整个过程呈现特殊的双峰型抑制效应。(中华眼底病杂志,2005,21:25-27)

Long-term effect of suramin on the inhibition of proliferation of cultured human retinal pigment epithelial cells

ObjectiveTo observe the long term effect of suramin on the inhibition of proliferation of human retinal pigment epithelial (RPE) cells in vitro. MethodsRPE cells grown in 9 pieces of 96 well plate (12 wells each plate) were divided into experimental and control group, with 6 wells in each group. The concentration of 0.1 ml RPE cells in each well is 5×10⁴ cells/ml. After the change of the medium, RPE cells were treated with suramin (250 μg/ml) in experimental group while treated with nothing in the control group. The medium of the 2 groups were changed to the normal medium after 4 days. At the 1^st, 2^nd, and 4^thday after the addition of suramin and at the 1^st, 2nd, 3^rd, 5^th, 6^th, 7^th, 9^th , 11^th and 13^th day after removing suramin, 1 plate was randomly selected to stop culturing, and the proliferation of RPE cells were detected by methyl thiazolyl tetrazolium (MTT) assay. ResultsUnder reversed microscope, RPE cells in control group were fused completely at the 7^th day after inoculation. The extracellular space of RPE cells in experimental groups was larger than that in the control group, and remained unfused at the 13th day after inoculation. The inhibitory rate of proliferation of RPE cells at the first day after treated with suramin was 14.85% and increased to the highest 25.79% at the 4th day. The first day after the suramin containing media was removed, the inhibitory rate decreased to 12.35%, and then raised gradually to over 20% at the 3rd to 5th day. Finally, the rate drop to 14.71%.ConclusionSuramin has the long-term effect on the inhibition of RPE cells induced by serum, especially the inhibitive effect after the remove of suramin, which indicates the specific double-peak inhibition during the whole process.(Chin J Ocul Fundus Dis, 2005,21:25-27)