Wright Catherine S, van Steensel Maurice A M, Hodgins Malcolm B, Martin Patricia E M
Department of Biological and Biomedical Sciences, Glasgow Caledonian University, UK.
Wound Repair Regen. 2009 Mar-Apr;17(2):240-9. doi: 10.1111/j.1524-475X.2009.00471.x.
Nonhealing cutaneous wounds, a major cause of morbidity and mortality, are difficult to treat. Recent studies suggest that significant increases in skin wound-healing rates occur by altering gap junction intercellular communication (GJIC). As migration of keratinocytes and fibroblasts is an important feature of wound healing, this study investigated whether migration rates in cultured normal human epidermal keratinocytes and dermal fibroblasts could be altered by modulating GJIC via connexin mimetic peptides. First, HeLa cells stably transfected with connexin43 (Cx43), Cx40, or Cx26 were used as a model to determine connexin specificity and the doses of connexin mimetic peptides required to attenuate GJIC. Gap26 and Gap26M inhibited GJIC dose dependently and were nonconnexin specific, whereas Gap27 was Cx43-selective. Skin keratinocytes and fibroblasts expressed a variety of connexins, with Cx43 predominating. Cx43 protein expression was reduced at leading edges 3 hours after scraping confluent monolayers, resolving at 24 hours. Gap26M and Gap27 significantly increased migration rates across scrapes in keratinocytes and fibroblasts by blocking gap junction functionality. GJIC inhibition can thus directly influence keratinocyte and fibroblast migration. Furthermore, our results support the therapeutic potential of connexin mimetic peptides to aid wound closure, and provide a simple approach to screening new agents.
难愈性皮肤伤口是发病和死亡的主要原因，治疗困难。最近的研究表明，通过改变缝隙连接细胞间通讯（GJIC）可显著提高皮肤伤口愈合率。由于角质形成细胞和成纤维细胞的迁移是伤口愈合的一个重要特征，本研究调查了通过连接蛋白模拟肽调节GJIC是否能改变培养的正常人表皮角质形成细胞和真皮成纤维细胞的迁移率。首先，将稳定转染连接蛋白43（Cx43）、Cx40或Cx26的HeLa细胞用作模型，以确定连接蛋白特异性以及减弱GJIC所需的连接蛋白模拟肽剂量。Gap26和Gap26M剂量依赖性地抑制GJIC，且不具有连接蛋白特异性，而Gap27具有Cx43选择性。皮肤角质形成细胞和成纤维细胞表达多种连接蛋白，其中Cx43占主导。在刮擦汇合单层细胞3小时后，Cx43蛋白表达在前沿降低，24小时后恢复。Gap26M和Gap27通过阻断缝隙连接功能显著提高角质形成细胞和成纤维细胞在刮擦处的迁移率。因此，GJIC抑制可直接影响角质形成细胞和成纤维细胞的迁移。此外，我们的结果支持连接蛋白模拟肽在促进伤口闭合方面的治疗潜力，并提供了一种筛选新药物的简单方法。
