Caffeic acid phenethylester increases stress resistance and enhances lifespan in Caenorhabditis elegans by modulation of the insulin-like DAF-16 signalling pathway.

Susannah Havermann,Yvonni Chovolou,Wim Wätjen,Hans-Ulrich Humpf,Junichi Sadoshima

doi:10.1371/journal.pone.0100256

Abstract

CAPE is an active constituent of propolis which is widely used in traditional medicine. This hydroxycinnamic acid derivate is a known activator of the redox-active Nrf2 signalling pathway in mammalian cells. We used C. elegans to investigate the effects of this compound on accumulation of reactive oxygen species and the modulation of the pivotal redox-active pathways SKN-1 and DAF-16 (homologues of Nrf2 and FoxO, respectively) in this model organism; these results were compared to the effects in Hct116 human colon carcinoma cells. CAPE exerts a strong antioxidative effect in C. elegans: The increase of reactive oxygen species induced by thermal stress was diminished by about 50%. CAPE caused a nuclear translocation of DAF-16, but not SKN-1. CAPE increased stress resistance of the nematode against thermal stress and finally a prolongation of the median and maximum lifespan by 9 and 17%, respectively. This increase in stress resistance and lifespan was dependent on DAF-16 as shown in experiments using a DAF-16 loss of function mutant strain. Life prolongation was retained under SKN-1 RNAi conditions showing that the effect is SKN-1 independent. The results of CAPE obtained in C. elegans differed from the results obtained in Hct116 colon carcinoma cells: CAPE also caused strong antioxidative effects in the mammalian cells, but no activation of the FoxO4 signalling pathway was detectable. Instead, an activation of the Nrf2 signalling pathway was shown by luciferase assay and western blots. CONCLUSION: CAPE activates the insulin-like DAF-16, but not the SKN-1 signalling pathway in C. elegans and therefore enhances the stress resistance and lifespan of this organism. Since modulation of the DAF-16 pathway was found to be a pivotal effect of CAPE in C. elegans, this has to be taken into account for the investigation of the molecular mechanisms of the traditional use of propolis.

Highlights

The hydroxycinnamic acid derivative caffeic acid phenethylester (CAPE, figure 1A) is a prominent component of propolis
Antioxidative capacity of CAPE We first estimated the antioxidative capacity of CAPE in a simple cell free system (TEAC assay: trolox equivalent antioxidative capacity assay)
To the best of our knowledge this is the first work that showed that pretreatment of C. elegans with CAPE strongly lowered the DCF fluorescence: In comparison to the DMSO control group the DCF

Summary

Introduction

The hydroxycinnamic acid derivative caffeic acid phenethylester (CAPE, figure 1A) is a prominent component of propolis (reviewed by [1]). G. antimicrobial, immunomodulatory, anticancer, hepatoprotective, neuroprotective and anti-inflammatory activities (reviewed by [2,3,4]). For this reason, propolis is recognised to have the potential for the development of new drugs (reviewed by [5]). Further anticancer effects of propolis and CAPE are reviewed by [9] and [10], respectively. To direct antioxidant properties [12] CAPE is known to activate the redox-sensitive Nrf pathway in mammalian cells, as shown e.g. renal epithelial cells [13] and astrocytes [14] and in a rat model in vivo [15]

Methods

Results

Conclusion