Transient receptor potential vanilloid-3 (TRPV3) channel induces dermal fibrosis via the TRPV3/TSLP/Smad2/3 pathways in dermal fibroblasts

J Dermatol Sci. 2020 Feb;97(2):117-124. doi: 10.1016/j.jdermsci.2019.12.011. Epub 2020 Jan 8.

Abstract

Background: Excessive wound healing can lead to hypertrophic scars, which are not only a cosmetic issue but could also be itchy or painful. Previously, we reported that, in comparison with normal tissue, thymic stromal lymphopoietin (TSLP) expression was increased in skin burn scars when the activation of transient receptor potential vanilloid-3 (TRPV3) of keratinocytes was increased. However, the functional role of TRPV3 in dermal fibrosis remains unclear.

Objective: We aimed to determine whether TRPV3 affects the collagen production of human primary dermal fibroblasts (HPDFs) and to investigate the mechanism involved.

Method: Human primary dermal fibroblasts were cultured and transformed into myofibroblasts using TSLP and carvacrol. Expression levels of α-SMA, fibronectin, and COL1A1 were determined using qPCR, western blotting, and immunofluorescence staining. Ca2+ influx was measured using a calcium-sensitive fluorescent dye, Fura3-AM. Nuclear factor of activated T-cells (NFAT) and phosphorylated-Smad2/3 were determined by western blotting. Silencing of TRPV3 with TRPV3-specific small interference RNA was evaluated using qPCR and western blotting.

Results: The expression levels of α-SMA, fibronectin, COL1A1, and TSLP were significantly increased in carvacrol-treated HPDFs. The expression levels of α-SMA, fibronectin, and COL1A1 were significantly increased by TSLP. The expression levels of TSLP and COL1A1 were significantly blocked by TRPV3 silencing in HPDFs. Regulation of Ca2+ influx and the expression levels of NFAT and p-Smad2/3 were significantly increased in carvacrol-treated HPDFs. ECM productions induced via the TRPV3/TSLP/Smad2/3 pathways.

Conclusions: The activation of the TRPV3 channels regulates dermal fibrosis by reducing extracellular matrix production via the TRPV3/TSLP/Smad2/3 pathways in dermal fibroblasts.

Keywords: Fibrosis; Thymic stromal lymphopoietin; Transient receptor potential vanilloid-3.

MeSH terms

  • Cell Differentiation / drug effects
  • Cells, Cultured
  • Collagen / biosynthesis
  • Cymenes / pharmacology
  • Cytokines / metabolism
  • Dermis / cytology
  • Dermis / pathology*
  • Extracellular Matrix / pathology*
  • Fibroblasts
  • Fibrosis
  • Humans
  • Primary Cell Culture
  • Signal Transduction / drug effects
  • Smad2 Protein / metabolism
  • Smad3 Protein / metabolism
  • TRPV Cation Channels / agonists
  • TRPV Cation Channels / metabolism*

Substances

  • Cymenes
  • Cytokines
  • SMAD2 protein, human
  • SMAD3 protein, human
  • Smad2 Protein
  • Smad3 Protein
  • TRPV Cation Channels
  • TRPV3 protein, human
  • TSLP protein, human
  • Collagen
  • carvacrol