Shao Yiming, Guo Zaiwen, Yang Yunxi, Liu Lu, Huang Jiamin, Chen Yi, Li Linbin, Sun Bingwei
Department of Burns and Plastic Surgery, Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou 215002, Jiangsu Province, China.
Burns Trauma. 2022 Nov 16;10:tkac044. doi: 10.1093/burnst/tkac044. eCollection 2022.
Inflammation is an important factor in pathological scarring. The role of neutrophils, one of the most important inflammatory cells, in scar hyperplasia remains unclear. The purpose of this article is to study the correlation between neutrophil extracellular traps (NETs) and scar hyperplasia and identify a new target for inhibiting scar hyperplasia.
Neutrophils were isolated from human peripheral blood by magnetic-bead sorting. NETs in plasma and scars were detected by enzyme-linked immunosorbent assays (ELISAs), immunofluorescence and flow cytometry. Immunohistochemistry was used to assess neutrophil (CD66B) infiltration in hypertrophic scars. To observe the entry of NETs into fibroblasts we used immunofluorescence and flow cytometry.
We found that peripheral blood neutrophils in patients with hypertrophic scars were more likely to form NETs ( < 0.05). Hypertrophic scars showed greater infiltration with neutrophils and NETs ( < 0.05). NETs activate fibroblasts to promote their differentiation and migration. Inhibition of NETs with cytochalasin in wounds reduced the hyperplasia of scars in mice. We induced neutrophils to generate NETs with different stimuli and detected the proteins carried by NETs. We did not find an increase in the expression of common scarring factors [interleukin (IL)-17 and transforming growth factor-β (TGF-β), > 0.05]. However, inhibiting the production of NETs or degrading DNA reduced the differentiation of fibroblasts into myofibroblasts. , NETs were found to be mediated by Toll-like receptor 9 (TLR-9) in fibroblasts and further phosphorylated nuclear factor Kappa-B (NF-κB). We found that IL-6, which is downstream of NF-κB, was increased in fibroblasts. Additionally, IL-6 uses autocrine and paracrine signaling to promote differentiation and secretion.
Our experiments found that NETs activate fibroblasts through the TLR-9/NF-κB/IL-6 pathway, thereby providing a new target for regulating hypertrophic scars.
炎症是病理性瘢痕形成的重要因素。中性粒细胞作为最重要的炎症细胞之一，在瘢痕增生中的作用尚不清楚。本文旨在研究中性粒细胞胞外诱捕网（NETs）与瘢痕增生之间的相关性，并确定抑制瘢痕增生的新靶点。
通过磁珠分选从人外周血中分离中性粒细胞。采用酶联免疫吸附测定（ELISA）、免疫荧光和流式细胞术检测血浆和瘢痕中的NETs。免疫组织化学用于评估肥厚性瘢痕中中性粒细胞（CD66B）的浸润情况。为观察NETs进入成纤维细胞的情况，我们采用了免疫荧光和流式细胞术。
我们发现肥厚性瘢痕患者外周血中性粒细胞更易形成NETs（<0.05）。肥厚性瘢痕显示中性粒细胞和NETs浸润更明显（<0.05）。NETs激活成纤维细胞以促进其分化和迁移。在伤口中用细胞松弛素抑制NETs可减少小鼠瘢痕增生。我们用不同刺激物诱导中性粒细胞产生NETs，并检测NETs携带的蛋白质。我们未发现常见瘢痕形成因子[白细胞介素（IL）-17和转化生长因子-β（TGF-β）]表达增加（>0.05）。然而，抑制NETs的产生或降解DNA可减少成纤维细胞向肌成纤维细胞的分化。发现NETs在成纤维细胞中由Toll样受体9（TLR-9）介导，并进一步磷酸化核因子κB（NF-κB）。我们发现成纤维细胞中NF-κB下游的IL-6增加。此外，IL-6通过自分泌和旁分泌信号促进分化和分泌。
我们的实验发现NETs通过TLR-9/NF-κB/IL-6途径激活成纤维细胞，从而为调节肥厚性瘢痕提供了新靶点。
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