Gynostemma pentaphyllum saponins induce melanogenesis and activate cAMP/PKA and Wnt/β-catenin signaling pathways

Phytomedicine. 2019 Jul;60:153008. doi: 10.1016/j.phymed.2019.153008. Epub 2019 Jul 1.

Abstract

Background: Melanogenesis is a physiological process of melanin production in response to UV exposure, which is modulated through multi-signaling pathways including cAMP/PKA, Wnt/β-catenin and MAPK signaling cascades.

Hypothesis/purpose: The present study aims to investigate the molecular mechanism of hyperpigmentation induced by Gynostemma pentaphyllum saponins.

Study design/methods: In this study, we investigated the melanogenic effects of triterpenoid saponins of Gynostemma pentaphyllum (GpS), a medicinal plant. Two mouse melanogenic cell lines B16 and B16F10 were employed for the current study.

Results: The results showed that non-toxic doses of GpS markedly increased melanin formation in both B16 and B16F10 cells. Western blot analysis showed that GpS treatment significantly up-regulated the expression levels of the key melanogenic proteins, including tyrosinase (TYR), microphthalmia-associated transcription factor (MITF), TRP-1 and TRP-2 in a dose-dependent manner. The phospho-CREB, which is the downstream target of PKA is also elevated upon GpS treatment. We further observed that H89, a PKA inhibitor, attenuated the GpS induced tyrosinase activity, melanin content, the expression of phospho-CREB. In addition to the cAMP/PKA signaling pathway, GpS treatment also up-regulated the β-catenin of the Wnt signaling pathway which is involved in the transcriptional activation of MITF in melanogensis. We further demonstrated that treatment with GpS markedly enhance mRNA expression of MITF, along with the downstream target molecules, TYR, TRP-1 and TRP-2. Knock-down MITF with siMITF inhibited the expression of MITF mRNA by 63%, and the melanin content was reduced 70% in the siMITF-transfected cells compared to untransfected or scramble siRNA control cells.

Conclusion: These findings demonstrated strong melanogenic activities of GpS, and the MITF is essential for the melanogenesis stimulated by GpS.

Keywords: Gynostemma pentaphyllum; Hypopigmentation; Melanogenesis; Triterpenoid saponins.

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Cyclic AMP / metabolism
  • Cyclic AMP-Dependent Protein Kinases / metabolism
  • Gene Knockdown Techniques
  • Gynostemma / chemistry*
  • Intramolecular Oxidoreductases / metabolism
  • Melanins / biosynthesis*
  • Melanoma, Experimental / metabolism*
  • Membrane Glycoproteins / metabolism
  • Mice
  • Microphthalmia-Associated Transcription Factor / metabolism
  • Monophenol Monooxygenase / metabolism
  • Oxidoreductases / metabolism
  • Saponins / pharmacology*
  • Up-Regulation
  • Wnt Signaling Pathway*

Substances

  • Melanins
  • Membrane Glycoproteins
  • Microphthalmia-Associated Transcription Factor
  • Saponins
  • Cyclic AMP
  • Oxidoreductases
  • Tyrp1 protein, mouse
  • Monophenol Monooxygenase
  • Cyclic AMP-Dependent Protein Kinases
  • Intramolecular Oxidoreductases
  • dopachrome isomerase