Mishima H, Hibino T, Hara H, Otori T
Department of Ophthalmology, Kinki University School of Medicine, Osaka-Sayama City, Osaka, Japan.
Curr Eye Res. 1996 Jul;15(7):733-8. doi: 10.3109/02713689609003456.
To understand the biological activity of entactin, a component of the basement membrane of the corneal epithelium, we investigated the ability of rabbit corneal epithelial cells to attach to an entactin matrix and the effect of entactin on the cells' attachment to other corneal basement proteins.
Multiwell plastic plates were coated with bovine serum albumin (BSA), alone or with BSA and entactin, laminin, fibronectin or collagen type IV. Cultured rabbit corneal epithelial cells were seeded on the plates. After incubation (usually 90 min), the cells were fixed and stained with 1% crystal violet. The number of attached cells was counted under a light microscope.
The numbers of attached cells increased in proportion to both the incubation period and the concentration of entactin coated. Furthermore, the number of cells attached to the entactin-coated plate was greater than the number attached to the BSA-coated plate for each incubation period (30 to 120 min). Likewise, when laminin-coated plates were treated with entactin, the number of the attached cells increased in proportion to the concentration of entactin. However, entactin did not affect the cellular attachment of fibronectin or type IV collagen. Cellular attachment to entactin was partially inhibited by the cells' preincubation with the synthetic peptide (GRGDSP).
The present results showed that cultured corneal epithelial cells adhere to entactin and that entactin stimulated the attachment of these cells to the laminin matrix. These findings suggest that entactin plays a specific role in maintaining the normal integrity of the corneal epithelium.
为了解角膜上皮基底膜成分巢蛋白的生物活性，我们研究了兔角膜上皮细胞附着于巢蛋白基质的能力以及巢蛋白对细胞附着于其他角膜基底蛋白的影响。
用牛血清白蛋白（BSA）单独或与巢蛋白、层粘连蛋白、纤连蛋白或IV型胶原一起包被多孔塑料板。将培养的兔角膜上皮细胞接种到板上。孵育（通常90分钟）后，固定细胞并用1%结晶紫染色。在光学显微镜下计数附着细胞的数量。
附着细胞的数量与孵育时间和包被的巢蛋白浓度均成比例增加。此外，在每个孵育时间（30至120分钟），附着于巢蛋白包被板的细胞数量均多于附着于BSA包被板的细胞数量。同样，当用巢蛋白处理层粘连蛋白包被的板时，附着细胞的数量与巢蛋白浓度成比例增加。然而，巢蛋白不影响纤连蛋白或IV型胶原的细胞附着。细胞与巢蛋白的附着被细胞与合成肽（GRGDSP）的预孵育部分抑制。
目前的结果表明，培养的角膜上皮细胞可附着于巢蛋白，且巢蛋白可刺激这些细胞附着于层粘连蛋白基质。这些发现提示巢蛋白在维持角膜上皮的正常完整性中起特定作用。
