Bazan H E, Tao Y, Bazan N G
Louisiana State University Eye Center, Louisiana State University Medical Center School of Medicine, New Orleans 70112.
Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8678-82. doi: 10.1073/pnas.90.18.8678.
Platelet-activating factor (PAF), a potent lipid mediator involved in inflammatory and immune responses, accumulates rapidly in response to injury in a variety of tissues, including the corneal epithelium. However, the precise role of this compound in the cascade of events following insult has not been defined. Here we examined the effect of PAF on gene expression in the epithelial cells of rabbit corneas in organ culture. We found that incubation with 100 nM methylcarbamoyl PAF, a nonhydrolyzable analog of PAF, produced rapid transient 2.8- and 3.5-fold increases in the expression of c-fos and c-jun, respectively, at 1 hr, followed by increased expression of the collagenase type I gene beginning at 3 hr and peaking at 14-fold by 8 hr. Addition of the protein-synthesis-inhibitor cycloheximide superinduced c-fos and c-jun, strongly potentiating the PAF effect, but inhibited the induction of collagenase type I expression, suggesting the existence of a transcriptional factor linking the two events. BN-50730, a selective antagonist of intracellular PAF-binding sites, blocked the expression of the immediate-early genes as well as the increase in collagenase type I mRNA. Our results suggest that one of the functions of PAF may be to enhance the breakdown of the extracellular matrix as a part of the remodeling process during corneal wound healing after injury. Pathologically, a PAF-induced overproduction of collagenase may be a factor in the development of corneal ulcers, as well as other pathophysiological conditions such as cartilage destruction in arthritis. If so, inhibitors of this signal-transduction pathway may be useful as tools for further investigation and, eventually, as therapeutic agents to treat such disorders.
血小板活化因子(PAF)是一种参与炎症和免疫反应的强效脂质介质,在包括角膜上皮在内的多种组织受到损伤时会迅速积累。然而,这种化合物在损伤后一系列事件中的精确作用尚未明确。在此,我们研究了PAF对器官培养的兔角膜上皮细胞基因表达的影响。我们发现,用100 nM甲基氨基甲酰PAF(PAF的一种不可水解类似物)孵育,在1小时时c-fos和c-jun的表达分别迅速短暂增加2.8倍和3.5倍,随后I型胶原酶基因的表达从3小时开始增加,到8小时时达到峰值,增加了14倍。添加蛋白质合成抑制剂环己酰亚胺可超诱导c-fos和c-jun,强烈增强PAF的作用,但抑制I型胶原酶表达的诱导,这表明存在一种连接这两个事件的转录因子。BN-50730是细胞内PAF结合位点的选择性拮抗剂,可阻断即刻早期基因的表达以及I型胶原酶mRNA的增加。我们的结果表明,PAF的功能之一可能是增强细胞外基质的分解,作为损伤后角膜伤口愈合过程中重塑过程的一部分。在病理情况下,PAF诱导的胶原酶过度产生可能是角膜溃疡以及其他病理生理状况(如关节炎中的软骨破坏)发生发展的一个因素。如果是这样,这种信号转导途径的抑制剂可能作为进一步研究的工具,最终作为治疗此类疾病的治疗剂而有用。