Department of Science and Environment, Roskilde University, Roskilde, Denmark.
Department of Biomedical Science, University of Copenhagen, Copenhagen, Denmark.
Wound Repair Regen. 2019 Nov;27(6):650-660. doi: 10.1111/wrr.12743. Epub 2019 Jul 9.
Antimicrobial peptides can have a dual role with both antimicrobial activity against a broad range of bacteria and immunomodulatory effect, making them attractive as therapeutic treatment of difficult wounds. Nisin A is widely known for its antimicrobial activity, and a preliminary study demonstrated that it increased wound closure, but the mechanism behind its effect is unknown. The aim of this study is to elucidate the wound healing potential of Nisin A and the mechanism behind. First, an epithelial and endothelial cell line, human keratinocyte (HaCaT) and human umbilical vein endothelial cell, were used to demonstrate migration and proliferation effects in vitro. From HaCaT cells and peripheral blood mononuclear cell, changes in cytokine levels were shown by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay. Second, the ex vivo porcine wound healing model was used to investigate the re-epithelization potential of Nisin A. Finally, the model Galleria mellonella was used to confirm antimicrobial activity and to investigate potential immunomodulatory effects in vivo. Here, we demonstrated that Nisin A affected migration significantly of both human umbilical vein endothelial cell and HaCaT cells (p < 0.05) but not proliferation, potentially by decreasing the levels of proinflammatory cytokines tumor necrosis factor-α, interleukin-6, and interleukin-8 (p < 0.001). Furthermore, Nisin A treatment diminished lipopolysaccharide-induced tumor necrosis factor-α levels from peripheral blood mononuclear cells and monocyte chemoattractant protein-1 from HaCaT cells (p < 0.001). Furthermore, Nisin A did not affect proliferation ex vivo either but increased re-epithelization of the porcine skin. Nisin A improved survival of G. mellonella significantly from Staphylococcus epidermidis (p < 0.001) but not from Escherichia coli, indicating that Nisin A did not help the larvae to survive the infection in a different than direct antimicrobial way. All together this makes Nisin A a potential treatment to use in wound healing, as it increases the mobility of skin cells, dampens the effect of lipopolysaccharide and proinflammatory cytokines, and decreases bacterial growth.
抗菌肽具有广谱抗菌活性和免疫调节作用的双重作用，因此作为治疗困难伤口的治疗方法具有吸引力。乳链菌肽 A 以其抗菌活性而广为人知，初步研究表明它可以增加伤口闭合，但尚不清楚其作用机制。本研究旨在阐明乳链菌肽 A 的伤口愈合潜力及其作用机制。首先，使用上皮细胞和成纤维细胞系，人角质形成细胞（HaCaT）和人脐静脉内皮细胞，在体外证明迁移和增殖作用。从 HaCaT 细胞和外周血单核细胞中，通过定量聚合酶链反应和酶联免疫吸附试验显示细胞因子水平的变化。其次，使用离体猪伤口愈合模型研究乳链菌肽 A 的再上皮化潜力。最后，使用模式昆虫大蜡螟来证实其体内的抗菌活性和潜在的免疫调节作用。在这里，我们证明乳链菌肽 A 显著影响人脐静脉内皮细胞和 HaCaT 细胞的迁移（p<0.05），但不影响增殖，这可能是通过降低促炎细胞因子肿瘤坏死因子-α、白细胞介素-6 和白细胞介素-8 的水平（p<0.001）。此外，乳链菌肽 A 处理可降低外周血单核细胞中脂多糖诱导的肿瘤坏死因子-α水平和 HaCaT 细胞中单核细胞趋化蛋白-1的水平（p<0.001）。此外，乳链菌肽 A 离体处理也不影响增殖，但增加了猪皮肤的再上皮化。乳链菌肽 A 显著提高了大蜡螟对表皮葡萄球菌的存活率（p<0.001），但对大肠杆菌没有帮助，这表明乳链菌肽 A 并没有以直接的抗菌方式帮助幼虫在感染中存活。所有这些都使乳链菌肽 A 成为伤口愈合的潜在治疗方法，因为它可以增加皮肤细胞的迁移能力，抑制脂多糖和促炎细胞因子的作用，并减少细菌生长。
