Hernnäs J, Särnstrand B, Lindroth P, Peterson C G, Venge P, Malmström A
Department of Physiological Chemistry 2, Lund University, Sweden.
Eur J Cell Biol. 1992 Dec;59(2):352-63.
Eosinophil cationic protein (ECP), a highly basic protein secreted from eosinophilic granulocytes, has been shown to take part in the inflammatory reaction. The involvement of ECP in fibroblast activation was therefore investigated in cell culture. Production of proteoglycans, hyaluronan and collagen in the presence of ECP was measured after incorporation of radioactive precursors and separation into different proteoglycan classes using gel and ion exchange chromatography and hydrophobic interaction chromatography. Proteoglycan accumulation in the cell layer was increased two- to fivefold at an ECP-concentration of 10 micrograms/ml. No effect on collagen, other proteins or hyaluronan was noted. Furthermore, no effect was observed on cell proliferation. The increased proteoglycan accumulation could be inhibited by addition of heparin or of antibodies to ECP. The effect could not be mimicked by the two basic peptides protamine and poly-L-lysine, speaking in favor of specificity. The increase in proteoglycan material was seen exclusively in the intracellular pool. No change of proteoglycans in the medium or the cell surface-associated pool was noted. The increase in the cell layer was accounted for by a two- to fivefold increase in free chains of heparan sulfate and dermatan sulfate. No change was seen in the proteoglycan pattern. No effect on proteoglycan synthesis or on endocytosis was noted. The increased accumulation of polysaccharide was caused by inhibited degradation of glycosaminoglycans. The half-lives of large and small heparan sulfate proteoglycans/glycosaminoglycans and dermatan sulfate proteoglycans/glycosaminoglycans in the cell layer are increased four- to sevenfold. We conclude that ECP inhibits proteoglycan degradation in fibroblasts, which indicates a role for the eosinophil in generation of fibrosis.
嗜酸性粒细胞阳离子蛋白（ECP）是一种由嗜酸性粒细胞分泌的高度碱性蛋白，已被证明参与炎症反应。因此，在细胞培养中研究了ECP对成纤维细胞活化的影响。在加入放射性前体后，使用凝胶、离子交换色谱和疏水相互作用色谱将其分离成不同的蛋白聚糖类别，然后测定在ECP存在下蛋白聚糖、透明质酸和胶原蛋白的产生。在ECP浓度为10微克/毫升时，细胞层中蛋白聚糖的积累增加了2至5倍。未观察到对胶原蛋白、其他蛋白质或透明质酸有影响。此外，未观察到对细胞增殖有影响。加入肝素或抗ECP抗体可抑制蛋白聚糖积累的增加。两种碱性肽鱼精蛋白和聚-L-赖氨酸无法模拟这种效果，这表明具有特异性。蛋白聚糖物质的增加仅在细胞内池中可见。未观察到培养基或细胞表面相关池中蛋白聚糖的变化。细胞层中的增加是由于硫酸乙酰肝素和硫酸皮肤素的游离链增加了2至5倍。蛋白聚糖模式未发生变化。未观察到对蛋白聚糖合成或内吞作用有影响。多糖积累的增加是由于糖胺聚糖降解受到抑制。细胞层中大分子和小分子硫酸乙酰肝素蛋白聚糖/糖胺聚糖以及硫酸皮肤素蛋白聚糖/糖胺聚糖的半衰期增加了4至7倍。我们得出结论，ECP抑制成纤维细胞中蛋白聚糖的降解，这表明嗜酸性粒细胞在纤维化形成中起作用。
