CBD Rewires the Liver Transcriptome and Calms Inflammatory Cytokines in Mice

Cannabidiol, the non-intoxicating compound extracted from Cannabis sativa, has been marketed and studied as a gentle anti-inflammatory, but the molecular reality of what it actually does inside the body has remained stubbornly vague. A new study published in Molecular Genetics and Genomics offers one of the most detailed pictures yet, mapping how CBD changes both the chemical messengers circulating in blood and the gene activity of two immune-critical organs at the same time. The researchers, led by Jakub Żurowski and Artur Gurgul at the University of Agriculture in Kraków, found that CBD does not simply switch inflammation off. Instead, it appears to act as a systemic immunometabolic modulator, coordinating changes across metabolism, circadian biology and immune signaling in a way that depends heavily on both dose and duration of exposure.

The team worked with male C57BL/6J mice, giving them purified CBD intraperitoneally at 0.2, 10 or 20 milligrams per kilogram of body weight. One group received the compound for just two days, representing short-term or sporadic use, while another group was treated daily for 28 days to mimic chronic exposure. Vehicle-treated controls received the same saline and Tween 80 solution without the active compound. The researchers then measured three classic pro-inflammatory cytokines in plasma, interleukin-1 beta, interleukin-6 and tumour necrosis factor alpha, using ELISA assays, and performed RNA sequencing on the liver and spleen to capture the full transcriptional response in each organ. Sequencing produced an average of 32.4 million raw reads per liver sample, with roughly three quarters of filtered reads uniquely mapped to the mouse reference genome GRCm39.

The cytokine results revealed a striking temporal split. After only two days of treatment, CBD significantly reduced IL-1β across all doses, with concentrations falling from about 69.7 picograms per millilitre in controls to between 49 and 57 picograms per millilitre, an 18 to 30 percent drop. TNF-α fell even harder, declining by roughly 20 to 45 percent in a dose-dependent fashion. IL-6, however, barely moved during this early window. Its decline emerged only during prolonged treatment: by day 14, IL-6 was significantly reduced at every dose, and by day 28 the two higher doses had driven levels down by approximately 40 to 45 percent, from 45.3 to under 29 picograms per millilitre. TNF-α remained suppressed at day 28 as well, suggesting that some anti-inflammatory effects persist while others fade or shift character over time.

The liver, the organ that also handles CBD clearance, turned out to be the epicentre of transcriptional change. Short-term high-dose treatment altered 526 genes, of which nearly 90 percent were upregulated, and enrichment analysis pointed squarely at mitochondrial energy metabolism: oxidative phosphorylation, electron transport chain activity, ATP synthesis and ribosome biogenesis were all prominently represented. The lowest short-term dose affected 116 genes linked to steroid and mineralocorticoid biosynthesis, tissue morphogenesis and lipid transport, while the intermediate dose touched only 11 genes, a reminder that cannabinoids often produce nonlinear, dose-dependent effects. After 28 days, the picture changed again. Long-term treatment altered between 104 and 404 genes depending on dose, with enrichment shifting toward fatty acid and eicosanoid metabolism, arachidonic acid pathways, and, at the highest dose, circadian rhythm regulation including the molecular clock gene Nr1d1, which encodes the nuclear receptor REV-ERBα.

That circadian finding may be one of the most intriguing threads in the study. REV-ERBα is a core component of the cellular clock, and previous work has shown it directly modulates innate immune responses and cytokine production. Immune function in general runs on a diurnal rhythm, with leukocyte trafficking and cytokine secretion varying robustly across the day. The fact that prolonged high-dose CBD enriched circadian and rhythmic process categories, while simultaneously downregulating genes involved in glucocorticoid receptor signalling and responses to nutritional and metabolic stimuli, suggests the compound may be tapping into the deep regulatory circuitry that connects clocks, stress hormones and metabolism rather than merely dampening inflammatory pathways one by one.

Perhaps the most counterintuitive result came at the highest long-term dose, where upregulated liver genes were enriched for innate immune and inflammatory processes, including antimicrobial humoral immunity, interleukin-1-mediated signaling and neutrophil chemotaxis. On its face, this looks like inflammation ramping up. Yet circulating cytokines remained reduced at the same time. The authors interpret this as evidence that transcriptional activation of immune-related pathways does not necessarily translate into systemic inflammatory output; it may instead reflect adaptive regulatory reprogramming. This distinction matters, because it separates genuine immunosuppression from a subtler recalibration of immune-metabolic networks, and it aligns with the well-established principle that cellular metabolism and immune cell behaviour are tightly intertwined.

The spleen told a very different story. Despite being the body’s central blood-filtering lymphoid organ, packed with T cells, B cells, macrophages and dendritic cells, and rich in CB2 cannabinoid receptors, the spleen showed remarkably modest transcriptional responses. Short-term treatment at the highest dose altered 40 genes, 38 of them downregulated, with the only significant enrichment involving lipid modification processes. Long-term treatment changed just two to four genes depending on dose. This muted splenic response, set against the liver’s dramatic remodeling, led the authors to a provocative conclusion: the systemic cytokine changes observed in blood are probably not driven primarily by classical immune organs, but rather by indirect regulation flowing from metabolically active tissues, with the liver acting as a kind of immunometabolic command centre.

Mechanistically, the findings fit CBD’s known multimodal pharmacology. Unlike THC, CBD has low affinity for the canonical CB1 and CB2 cannabinoid receptors and instead influences a scatter of molecular targets, including PPARγ, TRPV1 channels, adenosine signalling and redox-sensitive pathways, while inhibiting the transcription factor NF-κB, a master regulator of TNF-α and IL-1β expression. The early cytokine drops are consistent with NF-κB inhibition, while the delayed IL-6 decline may reflect a gradual attenuation of upstream inflammatory stimuli, since IL-6 production often sits downstream of IL-1β and TNF-α cascades. The lipid metabolism enrichment during long-term treatment echoes recent human cell studies showing that CBD can switch lipid mediator production toward inflammation-resolving species, and the authors’ own prior work found similar circadian and metabolic signatures in the kidneys of CBD-treated mice.

The study’s temporal dimension carries practical weight. Short-term exposure produced acute bioenergetic and stress-related transcriptional responses alongside rapid IL-1β and TNF-α suppression, whereas prolonged exposure shifted toward lipid handling, circadian regulation and broader immunometabolic pathways, with sustained IL-6 reduction. CBD’s high lipophilicity means it accumulates in lipid-rich, well-perfused tissues over repeated dosing, so the biological response evolves rather than simply intensifying. For anyone interpreting CBD experiments, or weighing its therapeutic potential, duration of exposure may matter as much as dose, and the two interact in ways that a single time point cannot capture.

The authors are careful about limitations. The pathways identified by RNA sequencing were not validated with targeted functional assays, bulk transcriptomics cannot distinguish changes within individual cell types from shifts in cellular composition, and only male mice were studied, leaving open the question of sex-dependent responses in endocannabinoid signalling and immunity. Still, the integrated picture is compelling: CBD lowered circulating pro-inflammatory cytokines while triggering strong, dose- and time-dependent transcriptional reprogramming in the liver and only limited changes in the spleen. Rather than acting as a blunt immunosuppressant, the compound appears to coordinate adjustments across mitochondrial bioenergetics, lipid metabolism, glucocorticoid pathways and circadian regulators such as REV-ERBα, nudging immune-metabolic networks toward balance. If that model holds up in functional studies and in both sexes, it could reshape how researchers think about CBD’s promise in inflammatory, autoimmune and metabolic disease.

Subject of Research: Effects of cannabidiol on systemic cytokines and liver and spleen transcriptomes in male mice

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

Article References: Żurowski, J., Ocłoń, E., Jasielczuk, I., Szmatoła, T., Mizera-Szpilka, K., Semik-Gurgul, E., Sawicki, S., & Gurgul, A. (2026). Cannabidiol (CBD) differentially shapes systemic cytokines and transcriptomic profiles in the liver and spleen: insights from a male mouse model. Molecular Genetics and Genomics, 301(1), Article 200. https://doi.org/10.1007/s00438-026-02525-w

Image Credits: AI Generated

DOI: 10.1007/s00438-026-02525-w

Keywords: cannabidiol, CBD, cytokines, transcriptomics, liver, spleen, immunometabolism, NF-kappaB, circadian rhythm, REV-ERBalpha, mouse model, inflammation