Insights into the mechanism of Piper betle leaf-induced contact leukomelanosis using C57BL/6 mice as the animal model and tyrosinase assays

Australas J Dermatol. 2011 Aug;52(3):172-8. doi: 10.1111/j.1440-0960.2010.00724.x. Epub 2011 Jan 12.


Background/objectives: Steamed piper betle leaves (PBL) were once used by many Taiwanese women to treat pigment disorders on the face. Most women claimed a quick, favourable response at first, only to be overcome with facial leukomelanosis later.

Methods: C57BL/6 mice were randomly assigned to different groups to study if PBL could cause the following effects: contact dermatitis, leukomelanosis, or hair bleaching. Intracellular melanin content was measured by tyrosinase assays.

Results: Most steamed PBL-treated mice developed contact dermatitis and postinflammatory hyperpigmentation (PIH) on their shaved backs. About half developed bleached hair to varying extents. The steamed PBL did not only bleach the hairs, but also, unexpectedly, stimulated melanocyte replication, indicated by the fact that the number of functional melanocytes in the tail epidermis increased significantly after treatment (P = 0.007). Using tyrosinase assays PBL extract at the undiluted concentration showed limited inhibition of melanogenesis, probably via melanocytotoxicity.

Conclusions: The leukomelanosis observed in patients might be the consequence of PIH combined with a mixed reaction (hyper- and hypopigmentation), probably due to the different volatile chemicals that surface after steaming the PBL. This conflicting mixed reaction suggests that counteractive ingredients might exist in PBL. PBL, if purified, might be a promising source of a novel bleaching agent.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Dermatitis, Irritant / etiology*
  • Dermatitis, Irritant / pathology
  • Female
  • Hair Diseases / chemically induced
  • Hair Diseases / pathology
  • Hyperpigmentation / chemically induced*
  • Hyperpigmentation / pathology
  • Hypopigmentation / chemically induced*
  • Melanins / analysis
  • Melanocytes / pathology
  • Mice
  • Mice, Inbred C57BL
  • Piper betle / toxicity*
  • Plant Extracts / pharmacology
  • Plant Extracts / toxicity*
  • Plant Leaves
  • Random Allocation


  • Melanins
  • Plant Extracts