Hyperglycaemic conditions decrease cultured keratinocyte mobility: implications for impaired wound healing in patients with diabetes
Hyperglycaemic conditions decrease cultured keratinocyte mobility: implications for impaired wound healing in patients with diabetes
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DOI：
10.1111/j.1365-2133.2008.08789.x
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发表时间：
2008-11-01
影响因子：
10.3
通讯作者：
Wu, C-S.
中科院分区：
文献类型：
--
作者：
Lan, C-C. E.;Liu, I-H.;Wu, C-S.
背景糖尿病（DM）患者血糖升高可导致包括伤口愈合不良在内的并发症。角质形成细胞的迁移和增殖是表皮再生的关键步骤。我们推测，受损的伤口愈合DM患者是由于中断适当的re-epithelialization.Objectives我们的目的是探讨高糖对角质形成细胞的细胞迁移和增殖方面的影响。用MTS法测定细胞活力。采用Transwell迁移实验和体外划痕实验评价其迁移率。检测基质金属蛋白酶（MMP）-2和MMP-9的mRNA表达及活性。还评估了它们各自的生理抑制剂，MMP（TIMP）-1和TIMP-2的组织抑制剂的mRNA。采用免疫荧光染色和蛋白质印迹法检测磷酸化粘着斑激酶（pp 125（FAK））的表达。用5-溴-2 '-脱氧尿苷掺入法检测高糖对角质形成细胞增殖的影响。与此相反，角质形成细胞的迁移率，MMP-2和MMP-9的活性和基因表达，pp 125（FAK）的表达，并在5天后的细胞增殖显着下调高血压治疗后，而TIMP-1的mRNA表达增加。结论在高血压条件下，角质形成细胞表现出减少迁移和增殖能力下降。角质形成细胞功能的这些损伤可能导致上皮再形成不足。这些有缺陷的生理事件为DM患者中常见的伤口愈合不良提供了合理的解释。
Background Elevated blood glucose in patients with diabetes mellitus (DM) leads to complications including poor wound healing. Proper keratinocyte migration and proliferation are the crucial steps during re-epithelialization. We hypothesize that the impaired wound healing in patients with DM is due to the disruption of proper re-epithelialization.Objectives We aimed to explore the effects of high glucose on keratinocytes in terms of cell migration and proliferation.Methods Keratinocytes were cultivated in normal and high glucose conditions. Their viability was evaluated by MTS assay. Transwell migration and in vitro scratch assays were used to evaluate their mobility. The mRNA expressions and activities of matrix metalloproteinase (MMP)-2 and MMP-9 were determined. The mRNA of their respective physiological inhibitors, tissue inhibitor of MMP (TIMP)-1 and TIMP-2, was also evaluated. Immunofluorescent staining and Western blotting were used to examine the expression of phosphorylated focal adhesion kinase (pp125(FAK)). The impacts of high glucose on keratinocyte proliferation were assessed by 5-bromo-2'-deoxyuridine incorporation assay.Results High glucose treatment did not affect keratinocyte viability up to 3 days. In contrast, the mobility of keratinocytes, the activities and gene expressions of MMP-2 and MMP-9, the expression of pp125(FAK), and the cell proliferation after 5 days were significantly downregulated after hyperglycaemic treatments while the mRNA expression of TIMP-1 increased.Conclusions Under hyperglycaemic conditions, keratinocytes demonstrate reduced migration and decreased proliferation capacities. These impairments of keratinocyte functions are likely to result in inadequate re-epithelialization. These defective physiological events provide a reasonable explanation for the poor wound healing commonly observed in patients with DM.