ALA-PDT promotes IL-1β secretion from human SZ95 sebocytes via activation of the NLRP3 inflammasome

Photodiagnosis Photodyn Ther. 2024 Apr:46:104007. doi: 10.1016/j.pdpdt.2024.104007. Epub 2024 Feb 7.

Abstract

Background: 5-Aminolevulinic acid photodynamic therapy (ALA-PDT) is an effective treatment for pilosebaceous inflammatory diseases, such as acne vulgaris. In this study, we explored ALA-PDT's mechanisms against acne in vitro.

Methods: We treated human SZ95 sebocytes with ALA (0.2 mM) and subjected them to varied PDT doses (0, 5, 10, 20 J/cm²) over 12 h. We assessed cell viability post-treatment using the Annexin V FITC/PI apoptosis kit. ROS accumulation in the sebocytes was detected with a DCFDA probe. We quantified NLRP3 and caspase-1 mRNA via quantitative PCR and determined IL-1β release following ALA-PDT by ELISA. Western blotting helped identify the levels of proteins associated with pyroptosis (NLRP3, caspase-1, and IL-1β). To elucidate the mechanisms, we re-evaluated these parameters after administering various concentrations of NAC antioxidants (0, 0.4, 2, 10 mM) and the caspase inhibitor Z-VAD-FMK (0, 5, 10, 20 μM).

Results: Increasing PDT dose inversely affected SZ95 sebocyte survival, with a corresponding rise in ROS and pyroptosis-related proteins (NLRP3, caspase-1, and IL-1β). Furthermore, NAC and Z-VAD-FMK modulated the expression and secretion of these molecules in a dose-responsive manner.

Conclusion: Our findings suggest ALA-PDT's potential mechanism of action on sebaceous glands could involve ROS induction, leading to NLRP3 inflammasome assembly, thereby heightening caspase-1 activation and IL-1β secretion. This cascade may amplify the local inflammatory response to break chronic inflammation in acne vulgaris treatment.

Keywords: ALA-PDT; Inflammasome NLRP3; Pyroptosis; SZ95 sebocytes.

MeSH terms

  • Acne Vulgaris / drug therapy
  • Aminolevulinic Acid* / pharmacology
  • Caspase 1 / metabolism
  • Cell Line
  • Cell Survival* / drug effects
  • Dose-Response Relationship, Drug
  • Humans
  • Inflammasomes* / drug effects
  • Inflammasomes* / metabolism
  • Interleukin-1beta* / drug effects
  • Interleukin-1beta* / metabolism
  • NLR Family, Pyrin Domain-Containing 3 Protein* / metabolism
  • Photochemotherapy* / methods
  • Photosensitizing Agents* / pharmacology
  • Pyroptosis / drug effects
  • Reactive Oxygen Species* / metabolism
  • Sebaceous Glands / drug effects

Substances

  • Aminolevulinic Acid
  • Caspase 1
  • Inflammasomes
  • Interleukin-1beta
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • NLRP3 protein, human
  • Photosensitizing Agents
  • Reactive Oxygen Species