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精氨酸-甘氨酸-天冬氨酸(RGD)肽对人晶状体上皮细胞系细胞黏附和迁移的抑制作用

Inhibitory effects of Arg-Gly-Asp (RGD) peptide on cell attachment and migration in a human lens epithelial cell line.

作者信息

Oharazawa Hideaki, Ibaraki Nobuhiro, Ohara Kunitoshi, Reddy Venkat N

机构信息

Department of Ophthalmology, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo, Japan.

出版信息

Ophthalmic Res. 2005 Jul-Aug;37(4):191-6. doi: 10.1159/000086595. Epub 2005 Jun 29.

Abstract

Posterior capsule opacification (PCO) after cataract surgery is caused by growth of residual human lens epithelial (HLE) cells on the posterior capsule. We have shown that extracellular matrix (ECM) is an essential factor for HLE cell attachment and migration. The purpose of this study was to examine the inhibitory effects of Arg-Gly-Asp (RGD) peptide on cell attachment and migration in an HLE cell line. HLE cell line cells (SRA 01/04) that were obtained by transfection of large T antigen of SV40 were cultured in the absence of serum. The culture dishes were coated with type IV collagen, laminin or fibronectin, and Gly-Arg-Gly-Asp-Ser-Pro (GRGDSP) RGD peptide (0.1, 0.3, 1.0, 2.0 mg/ml) was added to the medium. The number of attached cells was counted after 90 min of incubation, and the inhibitory effects of GRGDSP RGD peptide on cell attachment were calculated. Cell attachment on the fibronectin-coated dishes was inhibited by GRGDSP RGD peptide at concentrations higher than 0.3 mg/ml; the inhibitory rate was 80% at a concentration of 2.0 mg/ml. The inhibition of cell attachment by GRGDSP RGD peptide on laminin-coated dishes appeared only at a concentration of 2.0 mg/ml, whereas no effects were observed on the type IV collagen-coated dishes. The inhibitory effects of GRGDSP RGD peptide on cell migration were measured in medium containing 2.0 mg/ml of GRGDSP RGD peptide after 1, 3, 5 and 7 days of culture. Cell migration was inhibited by GRGDSP RGD peptide from 1 day of culture on the fibronectin-coated dishes and from 5 days of culture on the laminin-coated dishes, whereas no effects were observed on the type IV collagen-coated dishes. GRGDSP RGD peptide inhibited cell attachment and migration on laminin and fibronectin that have RGD sequences. These data suggested that RGD peptide may have the potential to prevent PCO.

摘要

白内障手术后的后囊膜混浊(PCO)是由后囊膜上残留的人晶状体上皮(HLE)细胞生长所致。我们已经表明,细胞外基质(ECM)是HLE细胞附着和迁移的重要因素。本研究的目的是检测精氨酸 - 甘氨酸 - 天冬氨酸(RGD)肽对HLE细胞系中细胞附着和迁移的抑制作用。通过转染SV40大T抗原获得的HLE细胞系细胞(SRA 01/04)在无血清条件下培养。培养皿用IV型胶原、层粘连蛋白或纤连蛋白包被,并向培养基中添加甘氨酸 - 精氨酸 - 甘氨酸 - 天冬氨酸 - 丝氨酸 - 脯氨酸(GRGDSP)RGD肽(0.1、0.3、1.0、2.0 mg/ml)。孵育90分钟后计数附着细胞的数量,并计算GRGDSP RGD肽对细胞附着的抑制作用。GRGDSP RGD肽在浓度高于0.3 mg/ml时可抑制纤连蛋白包被培养皿上的细胞附着;在浓度为2.0 mg/ml时抑制率为80%。GRGDSP RGD肽对层粘连蛋白包被培养皿上细胞附着的抑制作用仅在浓度为2.0 mg/ml时出现,而在IV型胶原包被培养皿上未观察到作用。在培养1、3、5和7天后,在含有2.0 mg/ml GRGDSP RGD肽的培养基中测量GRGDSP RGD肽对细胞迁移的抑制作用。GRGDSP RGD肽从培养第1天起抑制纤连蛋白包被培养皿上的细胞迁移,从培养第5天起抑制层粘连蛋白包被培养皿上的细胞迁移,而在IV型胶原包被培养皿上未观察到作用。GRGDSP RGD肽抑制具有RGD序列的层粘连蛋白和纤连蛋白上的细胞附着和迁移。这些数据表明RGD肽可能具有预防PCO的潜力。

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