胰岛素样生长因子结合蛋白-6对RPE细胞增殖和迁移的影响

Effects of insulin-like growth factor binding protein-6 on proliferation and migration of RPE cell

目的探讨胰岛素样生长因子结合蛋白-6(insulin-like growth factor binding protein-6,IGFBP-6)对视网膜色素上皮(retinal pigmented epithelium,RPE)细胞增殖和迁移的影响。方法用IGF-Ⅱ(50mg·L-1)、PDGF(20mg·L-1)、VEGF(40mg·L-1)和TGF-β(4mg·L-1)干预ARPE-19细胞6h,以诱导细胞增殖,再加入不同浓度IGFBP-6(1mg·L-1、10mg·L-1、100mg·L-1、500mg·L-1、1000mg·L-1)作用24h、48h后,通过MTS实验观察OD值的变化,研究IGFBP-6对细胞增殖的影响。细胞划痕实验分为IGF-Ⅱ组(50mg·L-1)、IGF-Ⅱ+IGFBP-6组(含IGF-Ⅱ50mg·L-1和IGFBP-6500mg·L-1)、无血清培养液组的实验,细胞划痕24h、48h后,计算各组细胞移行愈合率的变化,从而研究IGFBP-6对细胞迁移的影响。结果 MTS比色法显示一定浓度的IGFBP-6(100mg·L-1、500mg·L-1、1000mg·L-1)可以抑制IGF-Ⅱ诱导的ARPE-19细胞增殖(P<0.05),各浓度IGFBP-6对PDGF、VEGF、TGF-β诱导的细胞增殖均无明显影响(均为P>0.05);细胞划痕实验显示,IGF-Ⅱ组、IGF-Ⅱ+IGFBP-6组、无血清培养液组的细胞移行愈合率[分别为(24h:43.91%±3.85%、29.76%±2.49%、26.12%±2.33%;48h:66.09%±1.67%、59.88%±3.43%、57.05%±2.49%)]差异均有统计学意义(均为P<0.05)。结论 IGFBP-6能够抑制IGF-Ⅱ诱导的RPE细胞增殖和迁移。

Objective To study the effects of insulin-like growth factor binding protein-6（IGFBP-6） on proliferation and migration of retinal pigment epithelial（RPE） cells.Methods ARPE-19 cells were cultured with IGF-Ⅱ（50 mg·L-1）,PDGF（20 mg·L-1）,VEGF（40 mg·L-1） and TGF-β（4 mg·L-1） for 6 hours to induce the cell proliferation,then IGFBP-6（1 mg·L-1,10 mg·L-1,100 mg·L-1,500 mg·L-1,1000 mg·L-1）were added.OD value was tested by MTS method after 24 hours and 48 hours to detect the effect of IGFBP-6 on cell proliferation.The cell scratch test was divided into IGF-Ⅱ（50 mg·L-1） group,IGFBP-6＋IGF-Ⅱgroup（IGFBP-6 500 mg·L-1,IGF-Ⅱ 50 mg·L-1） and serum free medium group,the wound healing rate was observed at 24 hours and 48 hours,respectively to detect the effect of IGFBP-6 on cell migration.Results The MTS method outcomes indicated that IGFBP-6（100 mg·L-1,500 mg·L-1,1000 mg·L-1）could inhibit the proliferation of ARPE-19 induced by IGF-Ⅱ（P0.05）,but not by PDGF,VEGF,TGF-β（all P0.05）.Would healing rate in IGF-Ⅱgroup,IGFBP-6＋IGF-Ⅱgroup,serum free medium group was（43.91±3.85）%,（29.76±2.49）%,（26.12±2.33）% at 24 hours and（66.09±1.67）%,（59.88±3.43）%,（57.05±2.49）% at 48 hours,there were statistical differences（all P0.05）.Conclusion IGFBP-6 can inhibit the proliferation and migration of ARPE-19 cells induced by IGF-Ⅱ.