基质糖基化对角质形成细胞迁移功能损害的机制

Mechanism of glycated matrix imparing epidermal keratinocyte migration

目的探讨糖尿病条件下大鼠创面愈合过程中角质形成细胞迁移功能受损的可能机制。方法选用SD大鼠6只，取背部表皮进行角质形成细胞培养；制作并鉴定糖基化基质模型；评价糖基化基质对正常大鼠角质形成细胞迁移功能的影响；测定糖基化基质对正常大鼠角质形成细胞粘附功能、培养12，24h黏附形态、伪足形成、微丝表达、整合素表达的影响。结果糖基化层黏连蛋白对细胞迁移较对照组有明显抑制（P〈0．05）；对细胞贴壁率无明显影响（P〉0．05），但抑制了细胞的伸展及伪足的伸出，干扰了微丝的聚集，降低了整合素的表达（P〈0．05）。结论正常大鼠皮肤角质形成细胞在糖基化基质上出现迁移功能受阻，这可能是与整合素信号传导出现障碍，引起整合素、肌动蛋白表达减少和微丝形成，从而引起丝状伪足和板状伪足形成减少有关。

Objective To explore possible mechanism of glycated matrix impairing keratinoctye migration during the course of wound healing of diabetic rats. Methods Keratinocytes from backs of six Sprague-Dawley rats were cultured for 2-3 generations. Glycated laminin model was made to evaluate the effect of glycosylation on epidermal keratinocyte migration. In the meantime, the effect of glycated matrix on keratinocyte migration, adhesion rate at 12 and 24 hours of culture, cell morphous as well as expressions of F-actin and integrinα3 was observed. Results The amount of migrating keratinocytes in Group C was significantly more than that in Group G, which confirmed that keratinocyte migration was obviously inhibited by glycated matrix （ P 〈 0.05 ）. Glycated laminin exerted no significant effect on adhesion rate （ P 〉 0.05 ） but inhibited extension and pseudopod of keratinocytes, interfered aggregation of the microfilament and depressed the expression of keratinocyte integrinα3 （ P 〈 0.05 ）. Conclusions Keratinocyte migration is inhibited by the glycated laminin, as may be that integrin signaling disorder leads to decreased expressions of integrin and actin and less microfilament and hence results in less lamellipodia and filopodia.