Poly I:C induces collective migration of HaCaT keratinocytes via IL-8

Abstract

BackgroundDelayed wound healing reduces the quality of life (QOL) of patients. Thus, understanding the mechanism of wound healing is indispensable for better management. However, the role of innate immunity in wound healing is thus far unknown. Recently the involvement of TLR3 in wound healing has been evaluated. The systemic administration of polyriboinosinic-polyribocytidylic acid (poly I:C ; a substitute for viral dsRNA and a ligand of toll-like receptor 3), enhances wound healing in vivo. The aim of this study is to improve our understanding of the link between innate immunity and human wound healing, particularly in re-epithelialization.ResultsThe present study showed that poly I:C significantly accelerated collective HaCaT cell migration in a scratch assay. Poly I:C also increased IL-8 and bFGF production, and anti-IL-8 antibodies significantly inhibited the migration caused by poly I:C. Human recombinant IL-8 also accelerated collective HaCaT cell migration. An immunofluorescence assay and enzyme-linked immunosorbent assay (ELISA) also revealed that poly I:C decreased E-cadherin protein levels and increased vimentin protein levels, and anti-IL-8 antibody reversed this effect. In contrast, nucleic/cytosolic protein ratios of Snail 1 were unchanged in all tested conditions.ConclusionOur findings demonstrated that poly I:C accelerated collective HaCaT cell migration via autocrine/paracrine secretions of IL-8 and the subsequent incomplete epithelial-mesenchymal transition (EMT). Our findings provide a new strategy for wound healing by regulating innate immune systems in re-epithelialization.

Polo-like kinase 4 mediates epithelial–mesenchymal transition in neuroblastoma via PI3K/Akt signaling pathway

“…Further researches have elucidated suppression of EMT and subsequent inhibition of cell proliferation and invasion 59 . Furthermore, EMT could be reversible even though mesenchymal to epithelial transition was often an incomplete process 60 – 62 . In our study, we found that downregulation of PLK4 depressed EMT program in NB cells, including upregulation of the epithelial marker E-cadherin, as well as downregulation of mesenchymal markers (N-cadherin, Slug, and vimentin).…”

Lactobacillus crispatus accelerates re‐epithelialization in vaginal epithelial cell line MS74

“…32 Briefly, MS74 cells were grown to confluence in 24-well microplate (Falcon, Corning, NY, USA) with 1 mL of the medium described above. 32 Briefly, MS74 cells were grown to confluence in 24-well microplate (Falcon, Corning, NY, USA) with 1 mL of the medium described above.…”

“…The scratch assay was performed as previously described. 32 Briefly, MS74 cells were grown to confluence in 24-well microplate (Falcon, Corning, NY, USA) with 1 mL of the medium described above. A linear scratch was made using a 2-mm-wide Cell Scratcher (Iwaki Glass, Chiba, Japan), and the wells were washed once with phosphate buffered saline (PBS).…”

Soluble Fraction from Lysates of Selected Probiotic Strains Differently Influences Re-Epithelialization of HaCaT Scratched Monolayer Through a Mechanism Involving Nitric Oxide Synthase 2

“…Even if further in vivo studies should be conducted to confirm their potential therapeutic use, it will be of interest to firstly verify the reproducibility of the observed effects on HaCat cell line, also on primary keratinocytes, as well as other cell types involved in skin wound healing process (i.e., fibroblasts, endothelial cells), focusing on the biomolecular mechanism behind the observed effects. In this regard, considering the role attributed to the pro-inflammatory cytokines IL-6 and IL-8 in the tissue repair process [44][45][46] as well their association with NOS2 expression in different cellular models [47][48][49][50][51], our preliminary results ( Figure S1, Supplementary Materials) show that S. thermophilus, L. plantarum, and L. acidophilus samples, able to positively modulate NOS2/NO system and stimulate scratched monolayer repair, significantly increased the secretion of these two cytokines by HaCat; conversely B. infantis, B. breve, and B. longum, which caused a reduction of NOS2 expression and activity, also led to decreased levels of IL-6 and IL-8. L. bulgaricus, also in this context, did show no effect.…”