The antimicrobial peptide pheromone Plantaricin A increases antioxidant defenses of human keratinocytes and modulates the expression of filaggrin, involucrin, β-defensin 2 and tumor necrosis factor-α genes

Exp Dermatol. 2012 Sep;21(9):665-71. doi: 10.1111/j.1600-0625.2012.01538.x. Epub 2012 Jun 29.

Abstract

Plantaricin A (PlnA) is a peptide with antimicrobial and pheromone activities. PlnA was synthesized chemically and used as a pure peptide or synthesized biologically using Lactobacillus plantarum DC400 co-cultured with Lactobacillus sanfranciscensis DPPMA174. Cell-free supernatant (CFS) was used as a crude PlnA preparation. As estimated using the 3-(4,5-dimethyl-2-yl)-2,5-diphenyltetrazolium bromide and the 2',7'-dichlorofluorescein diacetate assays, both PlnA preparations increased the antioxidant defenses of human NCTC 2544 keratinocytes. PlnA (10 μg/ml) had a higher activity than hyaluronic acid or 125 μg/ml α-tocopherol. Effects on the transcriptional regulation of filaggrin (FLG), involucrin (IVL), hyaluronan synthase (HAS2), human β-defensin-2 (HBD-2) and tumor necrosis factor-alpha (TNF-α) genes were assayed. Compared with the control, expression of the FLG gene in NCTC 2544 cells increased in cells treated with hyaluronic acid, 1 or 10 μg/ml PlnA. Compared with the control, the level of IVL gene expression increased in NCTC 2544 cells treated with 10 μg/ml PlnA. No significant difference was found between the level of the HAS2 gene expressed by control cells and cells treated with PlnA. Compared with chemically synthesized PlnA, the up-regulation of the HBD-2 gene by CFS was higher. Compared with the control, expression of TNF-α decreased in NCTC 2544 cells after treatment with 1 or 10 μg/ml of chemically synthesized PlnA. In contrast, the level of TNF-α was highest in the presence of 10 μg/ml CFS-PlnA. These findings suggest that the PlnA was positively sensed by human keratinocytes, promoting antioxidant defenses, barrier functions and antimicrobial activity of the skin.

MeSH terms

  • Analysis of Variance
  • Bacteriocins / pharmacology*
  • Cell Survival / drug effects
  • Cells, Cultured
  • Gene Expression Regulation / drug effects*
  • Glucuronosyltransferase / genetics
  • Glucuronosyltransferase / metabolism
  • Humans
  • Hyaluronan Synthases
  • Hyaluronic Acid / pharmacology
  • Intermediate Filament Proteins / genetics
  • Intermediate Filament Proteins / metabolism
  • Keratinocytes / drug effects
  • Keratinocytes / metabolism*
  • Lipopolysaccharides / pharmacology
  • Oxidative Stress / drug effects*
  • Protein Precursors / genetics
  • Protein Precursors / metabolism
  • Tumor Necrosis Factor-alpha / genetics
  • Tumor Necrosis Factor-alpha / metabolism
  • Up-Regulation / drug effects
  • alpha-Tocopherol / pharmacology
  • beta-Defensins / genetics
  • beta-Defensins / metabolism

Substances

  • Bacteriocins
  • DEFB4A protein, human
  • Intermediate Filament Proteins
  • Lipopolysaccharides
  • Protein Precursors
  • Tumor Necrosis Factor-alpha
  • beta-Defensins
  • filaggrin
  • plantaricin A
  • involucrin
  • Hyaluronic Acid
  • Glucuronosyltransferase
  • HAS2 protein, human
  • Hyaluronan Synthases
  • alpha-Tocopherol