Cannabidiol (CBD) differentially shapes systemic cytokines and transcriptomic profiles in the liver and spleen: insights from a male mouse model

Abstract Cannabidiol (CBD) is widely studied for its anti-inflammatory and immunomodulatory properties, yet its relationship with systemic cytokine responses and organ-level transcriptional changes remains incompletely defined. Here, we investigated the dose- and time-dependent effects of CBD on circulating pro-inflammatory cytokines and the transcriptomic profiles of the liver and spleen in male C57BL/6J mice. Animals received CBD intraperitoneally at 0.2, 10, or 20 mg/kg body weight for either 2 days (short-term, ST) or 28 days (long-term, LT), with vehicle-treated controls. Serum IL-1β, IL-6, and TNF-α were measured by ELISA, and RNA-Seq was performed on liver and spleen to characterize differential gene expression and enriched biological processes. CBD significantly reduced IL-1β and TNF-α after ST treatment across doses, whereas IL-6 decreased primarily during LT exposure, with the strongest reduction at day 14 and partial persistence to day 28 at higher doses. Transcriptomic responses were markedly organ- and regimen-dependent. The liver transcriptome exhibited pronounced, dose- and time-dependent remodeling, with the largest number of differentially expressed genes at 20 mg/kg in ST and broad changes across doses in LT. The gene enrichment analyses indicated modulation of mitochondrial and translational pathways after ST high-dose exposure, lipid and fatty acid metabolism during LT, as well as circadian-associated regulation at higher-dose LT treatment, with innate immune–associated signaling terms enriched at the highest dose. In contrast, the spleen displayed comparatively modest transcriptional changes, with limited differential expression in LT and a clearer response only after ST exposure to 20 mg/kg, where altered genes showed trends related to lipid-associated processes. Together, these findings demonstrate that CBD attenuates circulating pro-inflammatory cytokines while inducing strong, dose- and time-dependent hepatic transcriptional reprogramming and relatively limited splenic transcriptomic shifts. The integration of systemic and organ-specific data supports a model in which CBD acts as a systemic immunometabolic modulator rather than a simple immunosuppressant.