Direct Visualization of Artepillin C into Fibroblast Cells via CARS Microscopy

Year 2018, Volume: 1 Issue: 3 - 2nd Propolis Conference Special Issue, 60 - 60, 08.12.2018

Wallance M. Pazın Vita Solovyeva Tibebe Lemma Bjarke Jørgensen Carlos José L. Constantıno Jonathan Brewer

Abstract

Artepillin C, the major component of the
Brazilian green propolis, has become the subject of great biological interest
among all the secondary metabolites found in that propolis, especially due to
its anticancer property. Biophysical studies have shown that the presence of
two prenylated groups in the molecular structure of Artepillin C enhances its
affinity for lipophilic environment¹, which is in agreement with biological assays reported in the
literature, suggesting that Artepillin C is absorbed through cell membranes². The purpose of this study was to obtain for the first time a
direct visualization of Artepillin C internalization into fibroblast cells by
means of Coherence Anti-Stokes Raman Scattering (CARS), a powerful microscopy
technique with a combination of two pulsed laser beams which induces a
vibrational motion of a specific compound, generating an intrinsic image³. A detailed vibrational characterization of Artepillin C has been
carried out using Raman microspectroscopy and surface-enhanced Raman scattering
(SERS) in combination with quantum chemical calculation based on LANL2DZ/B3LYP
level of theory.  The success use of CARS
microscopy was confirmed by collecting Artepillin C images from dry and
hydrated films of the compound, while the intrinsic images of Artepillin C
internalized into the cells was compared to confocal fluorescence microscopy
images by using a cell-membrane staining compound. With effect, this study
shows for the first time a direct visualization of the major compound of green propolis
into the cells by means of CARS microscopy.

 

Acknowledgements: FAPESP
(2017/23426-4) and DaMBIC (University of Southern Denmark).

Keywords

Fibroblast Cells, CARS Microscopy, Brazilian green propolis

References

• 1. Pazin, W. M. et al. (2017) Interaction of Artepillin C with model membranes. Eur. Biophys. J. 46: 383–393.
• 2. Paulino, N. et al. 2008 Anti-inflammatory effects of a bioavailable compound, Artepillin C, in Brazilian propolis. Eur. J. Pharmacol. 587: 296–301.
• 3. Evans, C. L. & Xie, X. S. (2008) Coherent Anti-Stokes Raman Scattering Microscopy: Chemical Imaging for Biology and Medicine. Annu. Rev. Anal. Chem. 1: 883–909.