角膜上皮细胞与内源性层粘连蛋白和纤连蛋白的附着。

Corneal epithelial cell attachment with endogenous laminin and fibronectin.

作者信息

Ohji M, Mandarino L, SundarRaj N, Thoft R A

机构信息

Department of Ophthalmology, University of Pittsburgh, Pennsylvania.

出版信息

Invest Ophthalmol Vis Sci. 1993 Jul;34(8):2487-92.

PMID:8325754

Abstract

PURPOSE

To evaluate the role of endogenously produced laminin and fibronectin as well as the effect of exogenous laminin and fibronectin in the attachment of human corneal epithelial cells in vitro.

METHODS

Primary cultured human corneal epithelial cells labeled with 3H-thymidine were seeded onto plates coated with laminin or fibronectin, or onto uncoated bacteriologic plates. Attachment of cells was measured in the presence or absence of antisera against laminin or fibronectin, by counting radioactivity.

RESULTS

Human corneal epithelial cells attached to plates coated with human laminin or human fibronectin in a dose-dependent manner, with 69% and 50% of cells attached to the wells coated with 40 micrograms/ml of laminin and fibronectin, respectively (P < 0.001). The percentage of attachment to uncoated bacteriologic plates increased from 1.2% at 45 min of incubation to 6.7% at 90 min, 22.2% at 3 hr, and 40.1% at 6 hr of incubation. Cycloheximide, a protein synthesis inhibitor, completely inhibited cell attachment. Rabbit antiserum against human fibronectin reduced cell attachment to the uncoated plates to 67% of the control value (P < 0.01), whereas rabbit antiserum against human laminin decreased the attachment to 52% of the control (P < 0.01). A combination of these two antisera reduced cell attachment to 46% of the control (P < 0.01).

CONCLUSIONS

Endogenous laminin and fibronectin as well as exogenous laminin and fibronectin play significant roles in the attachment of human corneal epithelial cells in culture.

摘要

目的

评估内源性产生的层粘连蛋白和纤连蛋白的作用，以及外源性层粘连蛋白和纤连蛋白对人角膜上皮细胞体外黏附的影响。

方法

将用³H - 胸腺嘧啶核苷标记的原代培养人角膜上皮细胞接种到涂有层粘连蛋白或纤连蛋白的平板上，或未涂覆的细菌学平板上。通过计数放射性，在存在或不存在抗层粘连蛋白或纤连蛋白抗血清的情况下测量细胞黏附情况。

结果

人角膜上皮细胞以剂量依赖方式黏附于涂有人类层粘连蛋白或人类纤连蛋白的平板上，分别有69%和50%的细胞黏附于涂有40微克/毫升层粘连蛋白和纤连蛋白的孔中（P < 0.001）。未涂覆的细菌学平板上的黏附百分比在孵育45分钟时从1.2%增加到孵育90分钟时的6.7%、3小时时的22.2%和6小时时的40.1%。蛋白质合成抑制剂环己酰亚胺完全抑制细胞黏附。兔抗人纤连蛋白抗血清将细胞对未涂覆平板的黏附降低至对照值的67%（P < 0.01），而兔抗人层粘连蛋白抗血清将黏附降低至对照的52%（P < 0.01）。这两种抗血清的组合将细胞黏附降低至对照的46%（P < 0.01）。