Leung E W, Rife L, Smith R E, Kay E P
Doheny Eye Institute, Los Angeles, CA 90033, USA.
Mol Vis. 2000 Mar 7;6:15-23.
To investigate the extracellular matrix macromolecules found in Descemet's membrane and in retrocorneal fibrous membrane (RCFM), and to examine whether the corneal endothelium has the capacity to produce both basement and non-basement membrane phenotypes.
Rabbit corneas with and without RCFM were analyzed by immunofluorescence using antibodies to 8 different collagens (basement membrane collagens: types IV and VIII; fibrillar collagens: types I and III; interfibrillar collagens: type VI and two spliced variant forms of type XII and one anchoring fiber: type VII), proteoglycans (perlecan and decorin), (beta)ig-h3 and laminin-1.
Normal corneal endothelium stains positively for all of the tested collagen types except type VII collagen. On the other hand, Descemet's membrane reacts positively only to the type IV collagen antibody. When non-collagenous components in normal cornea were examined, corneal endothelium stained positively for perlecan, decorin, (beta)ig-h3 and laminin, whereas Descemet's membrane staining for these proteins was negative. When collagenous components of RCFM were examined, RCFM stained positively for all of the tested collagen types except type IV collagen. When non-collagenous components of RCFM were examined, RCFM demonstrated a strong positive staining with decorin, (beta)ig-h3 and laminin, while perlecan staining was weak.
These observations suggest that corneal endothelium is able to produce both basement membrane phenotypes and non-basement membrane, fibrillar phenotypes. This in vivo study confirms our in vitro model of endothelial mesenchymal transformation, in which corneal endothelial cells are transformed to fibroblasts that are responsible for fibrosis.
研究在Descemet膜和角膜后纤维膜(RCFM)中发现的细胞外基质大分子,并检查角膜内皮是否有能力产生基底膜和非基底膜表型。
使用针对8种不同胶原蛋白(基底膜胶原蛋白:IV型和VIII型;纤维状胶原蛋白:I型和III型;纤维间胶原蛋白:VI型以及XII型的两种剪接变体形式和一种锚定纤维:VII型)、蛋白聚糖(基底膜聚糖和核心蛋白聚糖)、βig-h3和层粘连蛋白-1的抗体,通过免疫荧光分析有或没有RCFM的兔角膜。
正常角膜内皮对除VII型胶原蛋白外的所有测试胶原蛋白类型均呈阳性染色。另一方面,Descemet膜仅对IV型胶原蛋白抗体呈阳性反应。当检查正常角膜中的非胶原蛋白成分时,角膜内皮对基底膜聚糖、核心蛋白聚糖、βig-h3和层粘连蛋白呈阳性染色,而Descemet膜对这些蛋白质的染色为阴性。当检查RCFM的胶原蛋白成分时,RCFM对除IV型胶原蛋白外的所有测试胶原蛋白类型均呈阳性染色。当检查RCFM的非胶原蛋白成分时,RCFM对核心蛋白聚糖、βig-h3和层粘连蛋白呈强阳性染色,而基底膜聚糖染色较弱。
这些观察结果表明,角膜内皮能够产生基底膜表型和非基底膜、纤维状表型。这项体内研究证实了我们的内皮间充质转化体外模型,其中角膜内皮细胞转化为负责纤维化的成纤维细胞。