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.