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Spatial expression of metallothionein, matrix metalloproteinase-1 and Ki-67 in human epidermal wounds treated with zinc and determined by quantitative immunohistochemistry: a randomised double-blind trial
European Journal of Cell Biology ( IF 6.6 ) Pub Date : 2020-12-25 , DOI: 10.1016/j.ejcb.2020.151147
Magnus S Ågren 1 , Lana Chafranska 2 , Jens Ole Eriksen 2 , Julie Lyng Forman 3 , Morten J Bjerrum 4 , Peter Schjerling 5 , Heidi F Larsen 6 , Elena Cottarelli 7 , Lars N Jorgensen 8 , Lise Mette Rahbek Gjerdrum 9
Affiliation  

Reepithelialisation is fundamental to wound healing, but our current understanding largely relies on cellular and animal studies. The aim of the present randomised double-blind three-arm controlled trial was to correlate genuine epidermal wound healing with key proteins and topical zinc treatment in humans. Sixty wounds were produced using deroofed suction blisters in 30 healthy volunteers and randomised to topical zinc sulphate (n = 20), placebo (n = 20), or control (n = 20) treatment for 4 days. All wounds with perilesional skin were processed for automatic immunostaining of paraffin tissue sections with monoclonal antibodies against Ki-67, metallothionein (MT) and matrix metalloproteinase (MMP)-1. Protein expression was quantified by automated digital image analysis. Epidermal Ki-67 and MT labelling indices were increased in keratinocytes in the neoepidermis (∼1.1 mm) and at the wound edge (0.5 mm) compared to normal skin. Increased MMP-1 immunostaining was restricted to the neoepidermis. MT was robustly upregulated in the upper dermis of the wounds. Zinc treatment enhanced MMP-1 expression beneath the neoepidermis via paracrine mechanisms and MT under the neoepidermis and in the nonepithelialised wound bed via direct actions of zinc as indicated by the induction of MT2A mRNA but not MMP-1 mRNA in cultured normal human dermal fibroblasts by zinc sulphate. The present human study demonstrates that quantitative immunohistochemistry can identify proteins involved in reepithelialisation and actions of external compounds. Increased dermal MT expression may contribute to the anti-inflammatory activities of zinc and increased MMP-1 levels to promote keratinocyte migration.



中文翻译:

金属硫蛋白、基质金属蛋白酶-1 和 Ki-67 在锌治疗的人类表皮伤口中的空间表达并通过定量免疫组织化学测定:一项随机双盲试验

再上皮化是伤口愈合的基础,但我们目前的理解主要依赖于细胞和动物研究。本随机双盲三臂对照试验的目的是将真正的表皮伤口愈合与人类的关键蛋白质和局部锌治疗相关联。在 30 名健康志愿者中使用去顶吸水泡产生 60 个伤口,并随机接受局部硫酸锌 (n = 20)、安慰剂 (n = 20) 或对照 (n = 20) 治疗 4 天。对所有病灶周围皮肤的伤口进行处理,用针对 Ki-67、金属硫蛋白 (MT) 和基质金属蛋白酶 (MMP)-1 的单克隆抗体对石蜡组织切片进行自动免疫染色。通过自动数字图像分析量化蛋白质表达。与正常皮肤相比,新表皮 (~1.1 mm) 和伤口边缘 (0.5 mm) 角质形成细胞中的表皮 Ki-67 和 MT 标记指数增加。增加的 MMP-1 免疫染色仅限于新表皮。MT 在伤口的上层真皮中显着上调。锌处理通过旁分泌机制增强了新表皮下的 MMP-1 表达,通过锌的直接作用增强了新表皮下和非上皮伤口床中的 MMP-1 表达,如在培养的正常人真皮成纤维细胞中诱导 MT2A mRNA 而不是 MMP-1 mRNA 所示硫酸锌。目前的人体研究表明,定量免疫组织化学可以识别参与再上皮化和外部化合物作用的蛋白质。

更新日期:2020-12-25
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