The neuroprotective potential of
3,3'-diindolylmethane (DIM), which is a selective
aryl hydrocarbon receptor modulator, has recently been shown in cellular and animal models of
Parkinson's disease and
lipopolysaccharide-induced
inflammation. However, there are no data concerning the protective capacity and mechanisms of DIM action in neuronal cells exposed to
hypoxia. The aim of the present study was to investigate the neuroprotective potential of DIM against the
hypoxia-induced damage in mouse hippocampal cells in primary cultures, with a particular focus on DIM interactions with the
aryl hydrocarbon receptor (AhR), its nuclear translocator ARNT, and
estrogen receptor β (ERβ). In the present study, 18 h of
hypoxia induced apoptotic processes, in terms of the mitochondrial membrane potential, activation of
caspase-3, and fragmentation of cell nuclei. These effects were accompanied by substantial
lactate dehydrogenase release and neuronal cell death. The results of the present study demonstrated strong neuroprotective and anti-apoptotic actions of DIM in hippocampal cells exposed to
hypoxia. In addition, DIM decreased the Ahr and Arnt
mRNA expression and stimulated Erβ
mRNA expression level. DIM-induced
mRNA alterations were mirrored by changes in
protein levels, except for ERβ, as detected by ELISA, Western blotting, and immunofluorescence labeling. We also demonstrated that DIM decreased the expression of AhR-regulated
CYP1A1. Using specific siRNAs, we provided evidence that impairment of AhR and ARNT, but not ERβ plays a key role in the neuroprotective action of DIM against
hypoxia-induced cell damage. This study may have implication for identifying new agents that could protect neurons against
hypoxia by targeting AhR/ARNT signaling.