Involvement of cathepsin B in mitochondrial apoptosis by p-phenylenediamine under ambient UV radiation

J Hazard Mater. 2015 Dec 30:300:415-425. doi: 10.1016/j.jhazmat.2015.07.032. Epub 2015 Jul 18.

Abstract

Paraphenylenediamine (PPD), a derivative of paranitroaniline has been most commonly used as an ingredient of oxidative hair dye and permanent tattoos. We have studied the phototoxic potential of PPD under ambient ultraviolet radiation. PPD is photodegraded and form a novel photoproduct under UV A exposure. PPD shows a concentration dependent decrease in cell viability of human Keratinocyte cells (HaCaT) through MTT and NRU test. Significant intracellular ROS generation was measured by DCFDA assay. It caused an oxidative DNA damage via single stranded DNA breaks, micronuclei and CPD formation. Both lysosome and mitochondria is main target for PPD induced apoptosis which was proved through lysosomal destabilization and release of cathepsin B by immunofluorescence, real time PCR and western blot analysis. Cathepsin B process BID to active tBID which induces the release of cytochrome C from mitochondria. Mitochondrial depolarization was reported through transmission electron microscopy. The cathepsin inhibitor reduced the release of cytochrome C in PPD treated cells. Thus study suggests that PPD leads to apoptosis via the involvement of lysosome and mitochondria both under ambient UV radiation. Therefore, photosensitizing nature of hair dye ingredients should be tested before coming to market as a cosmetic product for the safety of human beings.

Keywords: Apoptosis; Cathepsin B; Cytochrome C; PPD (paraphenylenediamine); Phototoxicity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antioxidants / analysis
  • Antioxidants / metabolism
  • Apoptosis / drug effects*
  • Apoptosis / radiation effects*
  • Cathepsin B / antagonists & inhibitors
  • Cathepsin B / chemistry*
  • Cell Cycle / drug effects
  • Cell Line
  • DNA Damage
  • Humans
  • Keratinocytes / drug effects
  • Keratinocytes / radiation effects
  • Lysosomes / drug effects
  • Lysosomes / radiation effects
  • Membrane Potential, Mitochondrial / drug effects
  • Micronucleus Tests
  • Mitochondria / drug effects*
  • Mitochondria / radiation effects*
  • Phenylenediamines / pharmacology*
  • Reactive Oxygen Species / analysis
  • Ultraviolet Rays

Substances

  • Antioxidants
  • Phenylenediamines
  • Reactive Oxygen Species
  • Cathepsin B
  • 4-phenylenediamine