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胰岛素样生长因子结合蛋白-6在增生性玻璃体视网膜病变大鼠模型中的表达及其对视网膜色素上皮细胞增殖和迁移的影响。

Expression of IGFBP-6 in a proliferative vitreoretinopathy rat model and its effects on retinal pigment epithelial cell proliferation and migration.

作者信息

Zhao Hong-Mei, Sheng Min-Jie, Yu Jing

机构信息

Department of Ophthalmology, the Tenth People's Hospital of Tongji University, Shanghai 200072, China.

出版信息

Int J Ophthalmol. 2014 Feb 18;7(1):27-33. doi: 10.3980/j.issn.2222-3959.2014.01.05. eCollection 2014.

Abstract

AIM

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.

METHODS

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.

RESULTS

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)%].

CONCLUSION

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.

摘要

目的

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。

方法

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。

结果

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)%]。

结论

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。

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