Terashi Hiroto, Izumi Kenji, Deveci Mustafa, Rhodes Lenore M, Marcelo Cynthia L
Section of Plastic and Reconstructive Surgery, Department of Surgery, University of Michigan Medical Center, Ann Arbor, MI, USA.
Int Wound J. 2005 Dec;2(4):298-304. doi: 10.1111/j.1742-4801.2005.00148.x.
In order to more clarify the delayed wound healing in diabetes mellitus, we cultured the human epidermal keratinocytes in both 6 mM (control group) and 12 mM glucose (high-glucose group) of "complete" MCDB 153 medium. Hyperglycaemia slowed the rate of their proliferation and inhibited their DNA synthesis and the production of total proteins. By 1 month after primary seeding in high-glucose group, the cells ceased their proliferation, whereas the cells in control group grew for more than 40 days. Mean population doublings in high-glucose group was 5.27 (vs. 7.25 in control, P = 0.001), and mean population doubling time during 1 month in high glucose group was 5.43 days (vs. 3.65 days in control, P = 0.02). They indicate that prolonged exposure to high glucose decreases the replicative life span of human epidermal keratinocytes in vitro. Furthermore, analysis of fatty acid contents in membrane phospholipids with thin-layer and gas chromatography showed no difference between the cultured keratinocytes in both conditions. Immunocytochemical staining of glucose transporter 1 shows that 28.1% of cells in high-glucose group were almost twice positive of those in control group (13.2%, P = 0.008). The mechanism of the ill effects of high glucose on epidermal keratinocytes is not so far clear, but it indicates the possibility of any direct effect of hyperglycaemia on glucose metabolism without changing lipid metabolism on cell membrane. The high-glucose group presented in this report can be available as an in vitro valuable study model of skin epidermal condition on diabetes mellitus.
为了更清楚地了解糖尿病患者伤口愈合延迟的情况，我们在“完全”MCDB 153培养基的6 mM（对照组）和12 mM葡萄糖（高糖组）中培养人表皮角质形成细胞。高血糖减缓了它们的增殖速度，抑制了它们的DNA合成和总蛋白的产生。在高糖组原代接种后1个月，细胞停止增殖，而对照组的细胞生长超过40天。高糖组的平均群体倍增数为5.27（对照组为7.25，P = 0.001），高糖组1个月内的平均群体倍增时间为5.43天（对照组为3.65天，P = 0.02）。这些结果表明，长时间暴露于高糖环境会降低人表皮角质形成细胞在体外的复制寿命。此外，用薄层色谱法和气相色谱法分析膜磷脂中的脂肪酸含量，发现在两种条件下培养的角质形成细胞之间没有差异。葡萄糖转运蛋白1的免疫细胞化学染色显示，高糖组28.1%的细胞阳性率几乎是对照组（13.2%）的两倍（P = 0.008）。高糖对表皮角质形成细胞产生不良影响的机制目前尚不清楚，但这表明高血糖可能对葡萄糖代谢有直接影响，而不会改变细胞膜上的脂质代谢。本报告中呈现的高糖组可作为糖尿病皮肤表皮状况的一种有价值的体外研究模型。
