Non‐psychotropic Cannabis sativa L. phytocomplex modulates microglial inflammatory response through CB2 receptors‐, endocannabinoids‐, and NF‐κB‐mediated signaling

Abstract

Cannabis sativa L. is increasingly emerging for its protective role in modulating neuroinflammation, a complex process orchestrated among others by microglia, the resident immune cells of the central nervous system. Phytocannabinoids, especially cannabidiol (CBD), terpenes, and other constituents trigger several upstream and downstream microglial intracellular pathways. Here, we investigated the molecular mechanisms of a CBD‐ and terpenes‐enriched C. sativa extract (CSE) in an in vitro model of neuroinflammation. We evaluated the effect of CSE on the inflammatory response induced by exposure to lipopolysaccharide (LPS) in BV‐2 microglial cells, compared with CBD and β‐caryophyllene (CAR), CB2 receptors (CB2r) inverse and full agonist, respectively. The LPS‐induced upregulation of the pro‐inflammatory cytokines IL‐1β, IL‐6, and TNF‐α was significantly attenuated by CSE and only partially by CBD, whereas CAR was ineffective. In BV‐2 cells, these anti‐inflammatory effects exerted by CSE phytocomplex were only partially dependent on CB2r modulation and they were mediated by the regulation of enzymes responsible for the endocannabinoids metabolism, by the inhibition of reactive oxygen species release and the modulation of JNK/p38 cascade with consequent NF‐κB p65 nuclear translocation suppression. Our data suggest that C. sativa phytocomplex and its multitarget mechanism could represent a novel therapeutic strategy for neuroinflammatory‐related diseases.

A Novel Perilla frutescens (L.) Britton Cell-Derived Phytocomplex Regulates Keratinocytes Inflammatory Cascade and Barrier Function and Preserves Vaginal Mucosal Integrity In Vivo

“…Given the peculiar ability of PFP in modulating cytokines release induced by a 24 h inflammatory stimulus, we investigated whether this effect could be associated with an early regulation of MAPKs activity by measuring the phosphorylation levels of JNK in HaCaT cells stimulated with LCTM for 60 min through the ELISA dosage. As previously described [ 20 ], JNK activation peaked at 60 min with a significant increase of about 2-fold compared to control cells (one-way ANOVA: p = 0.0022 vs. CTRL) ( Figure 6 ). Statistical analysis revealed that treatment with PFP, at the dose of 100 µg/mL, was effective in counteracting significantly the LCTM-induced increased levels of JNK phosphorylation after 60 min (post hoc: p = 0.0019 vs. LCTM) ( Figure 6 ).…”

“…Cell viability was tested using Cell Counting kit-8 (CCK-8, Merck KgaA, Germany) as previously described [ 20 ]. Briefly, HaCaT and HFF cells (5 × 10 4 ) were cultured in 96-well plates.…”

“…MAPKs (JNK) activation was evaluated using non-competitive sandwich ELISA (Biolegend, Thermo Fisher Scientific, MA, USA) according to the manufacturer’s instructions as previously described by [ 20 ]; positive control furnished by the kit producer was used to validate the test.…”

Zingiber officinale Roscoe Rhizome Extract Exerts Senomorphic and Anti-Inflammatory Activities on Human Endothelial Cells

“…The concentrations of GIN, SHO, and ZTE were calculated by considering the percentage of each compound within the extract. Thus, the cells were pre-treated for 4 h with ZOE (10 μg/mL), GIN (1 μg/mL), SHO (0.17 µg/mL), and ZOE terpenoid-enriched fraction (ZTE, 3 µg/mL), and then stimulated with LPS (250 ng/mL) for 24 h [ 15 ].…”

Multi-Target Effects of ß-Caryophyllene and Carnosic Acid at the Crossroads of Mitochondrial Dysfunction and Neurodegeneration: From Oxidative Stress to Microglia-Mediated Neuroinflammation

“…BCP–CB 2 R interactions prevent LPS-induced oligodendrocyte toxicity and microglia imbalance [ 98 , 99 ], as well as C6 glioma cell excitotoxicity [ 100 ]. Moreover, a standardized non-psychotropic C. sativa extract containing BCP displays significant activity in reducing pro-inflammatory cytokine synthesis in activated BV-2 microglia cells through CB 2 R signalling [ 101 ]. In an experimental model of autoimmune encephalomyelitis, BCP was described as regulating local and systemic immunity [ 102 ].…”