Cannabinoid CB 1 and CB 2 receptors differentially regulate TNF-α-induced apoptosis and LPA 1-mediated pro-survival signaling in HT22 hippocampal cells

Aims: The aim of the study was to investigate the interaction between cannabinoid CB₁/CB₂ and lysophosphatidic acid (LPA) receptors in controlling neuronal signaling and fate.

Methods: HT22 hippocampal cells were treated with different cannabinoid and LPA receptor agonists and antagonists. Western blot and immunofluorescence microscopy were used to study intracellular signaling and the expression of apoptotic markers. Cell viability was determined by a luminescence assay.

Key findings: Cannabinoid agonists induced activation of both ERK1/2 and p38 MAP kinases. The effects of the CB₁/CB₂ receptor agonist HU210 were antagonized by the CB₁ antagonist rimonabant, whereas the responses to the CB₂ agonist JWH133 were blocked by the CB₂ antagonist SR144528. HU210 reduced the apoptotic cell death induced by the pro-inflammatory cytokine TNF-α, whereas JWH133 enhanced the cytokine cytotoxicity. Blockade of ERK1/2 and p38 MAPK activation abrogated the HU210 pro-survival and the JWH133 pro-apoptotic effects, respectively. HU210 and the endocannabinoid anandamide, but not JWH133, potentiated ERK1/2 stimulation by LPA and the tricyclic antidepressant amitriptyline acting through the LPA₁ receptor. HU210 enhanced amitriptyline-stimulated CREB phosphorylation and protection against TNF-α-induced apoptosis, whereas JWH133 had no effect. ERK1/2 stimulation by either HU210 or amitriptyline was dependent on fibroblast growth factor receptor (FGF-R) kinase activity and the combination of the two stimulants induced FGF-R phosphorylation. Moreover, the CB₁ receptor was found to co-immunoprecipitate with the LPA₁ receptor.

Conclusions: In HT22 hippocampal cells CB₁ and CB₂ receptors differentially regulate TNF-α-induced apoptosis and CB₁ receptors positively interact with amitriptyline-stimulated LPA₁ in promoting FGF-R-mediated ERK1/2 signaling and neuroprotection.