Zhao Hong-Mei, Sheng Min-Jie, Yu Jing
Department of Ophthalmology, the Tenth People's Hospital of Tongji University, Shanghai 200072, China.
Int J Ophthalmol. 2014 Feb 18;7(1):27-33. doi: 10.3980/j.issn.2222-3959.2014.01.05. eCollection 2014.
To investigate the expression of insulin-like growth factor binding protein-6 (IGFBP-6) in a proliferative vitreoretinopathy (PVR) model and its effects on proliferation and migration in retinal pigment epithelial (RPE) cells.
A PVR Wistar rat model was established by the intravitreal injection of RPE-J cells combined with platelet-rich plasma (PRP). The expression levels of IGFBP-6 were tested by ELISA. ARPE-19 cell proliferation was evaluated by the MTS method, and cell migration was evaluated by wound healing assays.
The success rate of the PVR model was 89.3% (25/28). IGFBP-6 was expressed at higher levels in the vitreous, serum and retina of rats experiencing advanced PVR (grade 3) than in the control group (vitreous: 152.80±15.08ng/mL vs 105.44±24.81ng/mL, P>0.05; serum: 93.48±9.27ng/mL vs 80.59±5.20ng/mL, P<0.05; retina: 3.02±0.38ng/mg vs 2.05±0.53ng/mg, P<0.05). In vitro, IGFBP-6 (500ng/mL) inhibited the IGF-II (50ng/mL) induced ARPE-19 cell proliferation (OD value at 24h: from 1.38±0.05 to 1.30±0.02; 48h: from 1.44±0.06 to 1.35±0.05). However, it did not affect basal or VEGF-, TGF-β- and PDGF-induced cell proliferation. IGFBP-6 (500ng/mL) reduced the IGF-II (50ng/mL)-induced would healing rate [24h: from (43.91±3.85)% to (29.76±2.49)%; 48 h: from (66.09±1.67)% to (59.88±3.43)%].
Concentrations of IGFBP-6 increased in the vitreous, serum, and retinas only in advanced PVR in vivo. IGFBP-6 also inhibited IGF-II-induced cell proliferation in a not dose or time dependent manner and migration. IGFBP-6 participates in the development of PVR and might play a protective role in PVR.
研究胰岛素样生长因子结合蛋白6（IGFBP-6）在增殖性玻璃体视网膜病变（PVR）模型中的表达及其对视网膜色素上皮（RPE）细胞增殖和迁移的影响。
通过玻璃体腔内注射RPE-J细胞联合富血小板血浆（PRP）建立PVR Wistar大鼠模型。采用酶联免疫吸附测定（ELISA）检测IGFBP-6的表达水平。采用MTS法评估ARPE-19细胞增殖，采用伤口愈合试验评估细胞迁移。
PVR模型成功率为89.3%（25/28）。晚期PVR（3级）大鼠玻璃体、血清和视网膜中IGFBP-6表达水平高于对照组（玻璃体：152.80±15.08ng/mL比105.44±24.81ng/mL，P>0.05；血清：93.48±9.27ng/mL比80.59±5.20ng/mL，P<0.05；视网膜：3.02±0.38ng/mg比2.05±0.53ng/mg，P<0.05）。在体外，IGFBP-6（500ng/mL）抑制IGF-II（50ng/mL）诱导的ARPE-19细胞增殖（24小时光密度值：从1.38±0.05降至1.30±0.02；48小时：从1.44±0.06降至1.35±0.05）。然而，它不影响基础或VEGF、TGF-β和PDGF诱导的细胞增殖。IGFBP-6（500ng/mL）降低了IGF-II（50ng/mL）诱导的伤口愈合率[24小时：从（43.91±3.85）%降至（29.76±2.49）%；48小时：从（66.09±1.67）%降至（59.88±3.43）%]。
体内仅在晚期PVR的玻璃体、血清和视网膜中IGFBP-6浓度升高。IGFBP-6还以非剂量或时间依赖性方式抑制IGF-II诱导的细胞增殖和迁移。IGFBP-6参与PVR的发生发展，并可能在PVR中发挥保护作用。
