Neuroprotective Effects of Sulforaphane and the Involvement of Autophagy

Bednar, Sandra (2016) Neuroprotective Effects of Sulforaphane and the Involvement of Autophagy. Doctoral thesis, University of East Anglia.

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Abstract

ABSTRACT
Sulforaphane (SFN) is an isothiocyanate found mostly after consumption of broccoli, but also other cruciferous vegetables such as cauliflower or cabbage. Isothiocyanates have been researched for over 20 years in the field of cancer. They show many different bioactivities, of which the majority are positively associated with cell health and are possibly collectively responsible for protective effects of SFN against toxin-induced cell death.
This study aims to investigate the bioactivities of SFN on neuronal cells. PC-12 and SH-SY5Y cells were used to resemble neuronal-like systems to research especially autophagy as well as nuclear factor E2-related factor 2 (Nrf2) and how these pathways might influence neuroprotective abilities of SFN.
Basal assessments confirmed that SFN can induce Nrf2-driven phase II enzymes, as determined by qPCR as well as immunoblotting. The elevation of the main autophagy marker light chain 3-II (LC3-II) by SFN could be observed dose-dependently at protein level. In addition, SFN pre-treatment provided statistically significant cytoprotection against H2O2- and 6-hydroxydopamine- (6-OHDA) induced cell death.
Further, DJ-1, a multifunctional protein, was selected for investigation with SFN, since it is highly implicated with neuronal cell health. SFN induced DJ-1 protein levels dose-dependently. In addition, tunicamycin-induced ER-stress was significantly reduced by SFN, as shown using the ER-stress marker CHOP on protein and RNA levels.
To intensify the research of SFN and autophagy, primary neuronal cells (PNCs) were developed from embryos from Atg16L1 wild type and knock out (KO) mice. Atg16L1 is a protein necessary for autophagosome formation. Immunostainings assessed that autophagy was indeed fully supressed in KO cells. Preliminary results suggest that autophagy is involved in the neuroprotective effects of SFN.
Conclusively, SFN was able to significantly protect all neuronal cells investigated from H2O2- or 6-OHDA-induced cell death. SFN’s ability to activate autophagy as well as DJ-1 may contribute to its protective effects.

Item Type: Thesis (Doctoral)
Faculty \ School: Faculty of Medicine and Health Sciences > Norwich Medical School
Depositing User: Vailele Chittock
Date Deposited: 15 Jun 2016 11:41
Last Modified: 15 Jun 2016 11:41
URI: https://ueaeprints.uea.ac.uk/id/eprint/59348
DOI:

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