舒拉明对碱性成纤维细胞生长因子促进人视网膜色素上皮细胞增殖作用拮抗效应的研究

The inhibiting effects of suramin on bFGF induced proliferation of cultured human RPE cells

目的探讨舒拉明抗增生性玻璃体视网膜病变(PVR)的作用及机制。方法采用体外培养的人眼视网膜色素上皮(RPE)细胞,总共10组。其中5个对照组的培养基中分别加入终浓度为1、10、100、500ng/ml的bFGF和3125μg/ml的舒拉明。实验组4组,培养基中除加入3125μg/ml的舒拉明外,还分别加入终浓度为1、10、100和500ng/ml的bFGF。另设1组空白组。培养48h后,采用MTT法,酶联免疫检测仪测定各孔吸光度(A)值,计算细胞的增殖率。采用析因分析比较bFGF和舒拉明的交互效应和单独效应。结果各组细胞生长无明显的形态学差异。舒拉明与bFGF存在拮抗性交互效应(F=673,P<001)。对照组比较空白组,全部bFGF组A值都增加,最大A值为bFGF的10ng/ml浓度组;舒拉明组则降低;实验组中,各组A值较bFGF对照组都下降,最大A值左移对应bFGF的100ng/ml浓度组。结论舒拉明能够拮抗bFGF促RPE细胞增殖的作用,其抑制RPE细胞增殖并进一步防治PVR的机制至少包括拮抗了生长因子的生物学效应。

Objective To study the inhibiting effects of suramin on FGF induced proliferation of cultured RPE cells and discuss the effects and mechanisms of suramin in the prevention of proliferative vitreoretinopathy (PVR). Methods Human RPE cells at passage 6 were seeded into 96-well plates. In the 5 control groups, different concentrations of bFGF (1, 10, 100 and 500 ng/ml) or suramin (31.25 μg/ml) were added into the culture medium. In the 4 treatment groups, 31.25 μg/ml suramin with different concentrations of bFGF (1, 10, 100 and 500 ng/ml) were added to the medium. The blank group was incubated with serum-free DMEM only. After incubated for 48 hours, the proliferation of cells in all groups was measured by the MTT assay and expressed as light absorption values. The separate and combine effects of suramin and bFGF were analysed. Results Under the phase-contrast microscope, no changes of cell morphology could be detected between various groups. Suramin showed an inhibitory effects on bFGF induced proliferation of RPE cells (F=6.73,P<0.01). Light absorption value (A value) in bFGF-treated control groups was higher than that of the blank group and reached to the peak in cells treated with ~10 ng/ml of bFGF; with no further increase in cells treated with higher levels of bFGF. The A value in suramin-treated control group was lower than that of the blank group. The A-value in all treated groups was lower than that of the control groups treated with various concentrations of bFGF. In the treated group, the highest A value was detected in cells treated with 100 ng/ml bFGF. With further increase of bFGF concentration to 500 ng/ml, the A-value was not statistically different from that of cells treated with 100 ng/ml bFGF.Conclusions^Suramin can inhibit the bFGF induced proliferation of RPE cells. The mechanisms of the effects of suramin in the inhibition of proliferation of RPE cells and in the prevention of PVR at least partly due to its competition effects with the growth factors. (Chin J Ophthalmol, 2005, 41:110-113)