EGF AND PGE(2) - EFFECTS ON CORNEAL ENDOTHELIAL-CELL MIGRATION AND MONOLAYER SPREADING DURING WOUND REPAIR IN-VITRO
EGF AND PGE(2) - EFFECTS ON CORNEAL ENDOTHELIAL-CELL MIGRATION AND MONOLAYER SPREADING DURING WOUND REPAIR IN-VITRO
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DOI：
10.3109/02713689508998408
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发表时间：
1995-07-01
影响因子：
2
通讯作者：
MEKLIR, B
中科院分区：
文献类型：
--
作者：
JOYCE, NC;JOYCE, SJ;MEKLIR, B
成人角膜内皮的体内修复主要是通过细胞进入创面缺损处而不是通过细胞分裂来实现的。两种形式的细胞运动有助于内皮损伤修复：单个细胞迁移到缺损区和融合单层扩散到创面区域。本实验室建立了用有丝分裂抑制剂5-氟尿嘧啶处理的兔角膜内皮细胞进行组织培养的模型，以模拟体内人角膜内皮细胞的相对无丝分裂状态。这个模型可以研究生长因子和其他因素对创伤后单个细胞迁移和单层扩散的影响。在培养的角膜内皮细胞中检测到表皮生长因子(EGF)及其受体的mRNA，并在原位检测到人角膜内皮细胞上的EGF受体，提示该生长因子可能以自分泌的方式发挥作用。前列腺素E(2)(PGE(2))是由培养的角膜内皮细胞合成的，在角膜损伤和前房炎症反应中以相对较高的数量存在于房水中。虽然角膜内皮细胞可能同时暴露于EGF和PGE(2)，但对它们对单层修复的影响知之甚少。本研究使用创伤模型系统比较了单独使用PGE(2)、单独使用EGF以及两种药物联合使用对单个细胞迁移和单层扩散的影响，并使用商业免疫分析方法确定了EGF对PGE(2)分泌的影响。外源前列腺素E(2)处理受伤细胞15分钟，可刺激单个细胞迁移并抑制单层扩散。在没有内源性或外源性PGE(2)的情况下，创伤培养物暴露于EGF 15min对两种形式的细胞运动都没有显著影响。在创伤培养中加入EGF+PGE(2)15min，可显著刺激单个细胞迁移，高于单独使用PGE(2)，但对高于单独使用PGE(2)的细胞迁移没有影响。免疫分析结果表明，培养的角膜内皮细胞合成和分泌PGE(2)。10 ng/mlEGF作用15min足以使PGE(2)分泌量增加56%，而EGF处理24 h则使PGE(2)分泌量增加260%。1MU的吲哚美辛可抑制原生性和EGF诱导的PGE(2)合成。结果提示，在角膜损伤或炎症时，EGF和PGE(2)可显著影响促进角膜内皮修复的细胞运动。
In vivo repair of the adult human corneal endothelium occurs mainly by movement of cells into the wound defect rather than by cell division. Two forms of cell movement contribute to endothelial wound repair: migration of individual cells into the defect and spreading of the confluent monolayer into the wound area. This laboratory has developed a tissue culture model using rabbit corneal endothelial cells pretreated with the mitotic inhibitor 5-fluorouracil to mimic the relatively amitotic state of human corneal endothelium in vivo. This model permits study of the effects of growth factors and other agents on individual cell migration and monolayer spreading in response to wounding. mRNA for epidermal growth factor (EGF) and its receptor has been detected in cultured corneal endothelial cells and EGF receptors have been detected on human corneal endothelial cells in situ, suggesting that this growth factor may act in an autocrine manner. Prostaglandin E(2) (PGE(2)) is synthesized by cultured corneal endothelial cells and is present in relatively high quantity in aqueous humor in response to corneal wounding and to inflammation in the anterior chamber. Although corneal endothelial cells may be exposed to both EGF and PGE(2), little is known about their effects on monolayer repair. The current study compared the effects of PGE(2) alone, EGF alone, and both agents in combination on individual cell migration and monolayer spreading using the wound model system and also determined the effect of EGF on PGE(2) secretion using a commercial immunoassay. A 15 min exposure of wounded cultures to exogenous PGE(2) stimulated individual cell migration and suppressed monolayer spreading. In the absence of either endogenous or exogenous PGE(2), exposure of wounded cultures to EGF for 15 min had no significant effect on either form of cell movement. Addition of EGF plus PGE(2) to wounded cultures for 15 min significantly stimulated individual cell migration above that observed with PGE(2) alone, but had no effect on spreading above that observed with PGE(2) alone. Immunoassay results indicated that cultured corneal endothelial cells constitutively synthesize and secrete PGE(2). A 15 min exposure of cultures to 10 ng/ml EGF was sufficient to increase PGE(2) secretion 56% above constitutive levels, while a 24 h treatment with EGF increased PGE(2) secretion 260% over constitutive levels. Both constitutive and EGF-induced PGE(2) synthesis were inhibited by 1 mu M indomethacin. Results suggest that, upon corneal wounding or inflammation, EGF and PGE(2) could significantly affect the cellular movements which contribute to corneal endothelial repair.