Phytocannabinoids reduce inflammation of primed macrophages and enteric glial cells Critical review of the effect of phytocannabinoids on gut immune cells. Phytocannabinoids reduce inflammation of primed macrophages and enteric glial cells Critical review of the effect of phytocannabinoids on gut immune cells

In vitro study analyzed the putative-anti-inflammatory effect of nine-selected pure cannabinoids on enteric glial cells (EGC’s) triggered to undergo inflammation with lipopolysaccharide (LPS) . Our results demonstrate that THC at the lower concentrations tested exerted the most effective anti- inflammatory effect in EGC’s compared to other phytocannabinoids tested herein. We then performed RNA-seq analysis of EGC’s exposed to LPS in the presence or absence of THC or THC-COOH. Transcriptomic analysis of these EGC’s revealed 23 differentially expressed genes (DEG) compared to treatment with only LPS. Pretreatment with THC resulted in 26 DEG and THC-COOH 25 DEG. To evaluate biological pathways affected by the different phytocannabinoid treatments we used the Ingenuity platform. We show that THC treatment but not THC-COOH affected Overall design: Enteric glial cell lines (EGC/PK060399egfr) were grown in 75 mm2 flasks to 70% confluence in DMEM medium containing 10% (Sigma Aldrich, USA) fetal bovine serum (FBS) (Biological industries), 0.5% penicillin-streptomycin (Bio-logical industries), at 37°C under 5% CO2. Cells were trypsinized (using 0.25%) (Invitro-gen, Carlsbad CA, USA) and transferred every 2-3 days. Cells were incubeted for 1h without treatment, with 0.1hg/mL pure THC, 10nM pure THC-COOH, 10ug/L Sparstolonin B (Ssnb) (positive control). After 1h 1ug/mL LPS (E. coli 0111:B4) was added for 24h. After 24h media was removed and RNA was extracted using MN NucleoSpin RNA Kit.