Estudo da modulação imunometabólica da Cannabis medicinal na obesidade

Abstract

Obesity is a multifaceted condition characterized by chronic inflammation, metabolic dysregulation, and an increased risk of severe complications. Current anti-obesity pharmacotherapies face significant challenges, including adverse side effects and high rates of weight regain after discontinuation. In this context, targeting chronic inflammation has emerged as a promising therapeutic strategy. Phytocannabinoids derived from Cannabis sativa, such as THCV, CBD, and CBG, have demonstrated potential in reducing inflammation through interactions with cannabinoid receptors and other signaling pathways. The immunomodulatory effects of various phytocannabinoid-rich extracts on chronic inflammation and metabolic alterations were evaluated in peripheral blood mononuclear cells (PBMCs) from obese individuals, to identify novel adjuvant therapeutic strategies. Comparative analyses were conducted on monocytes and total CD4+ T cells isolated from both eutrophic and obese volunteers. PBMCs were stimulated ex vivo with the phytocannabinoid-rich extract that exhibited the most promising activity. Positive controls comprised lipopolysaccharide (LPS) for monocyte activation and anti-CD3/CD28 antibodies for CD4+ T cell stimulation in flow cytometry assays, while dexamethasone and Staphylococcal enterotoxin B (SEB) served as anti-inflammatory and pro-inflammatory controls, respectively, in ELISA assays. Before the immunological tests, cytotoxicity of crude C. sativa extracts was assessed in murine RAW 264.7 macrophages, human erythrocytes, and PBMCs to establish safe experimental concentrations. Viability assays (MTT) revealed CC50 values in RAW 264.7 cells of 28.49 μg/mL for CBD, 47.56 μg/mL for CBG, and 8.68 μg/mL for THCV, with no hemolytic activity detected in erythrocytes. PBMC CC50 values were 5.91 μg/mL (CBD), 15.77 μg/mL (CBG), and 6.79 μg/mL (THCV). Annexin V/propidium iodide staining indicated apoptosis as an action mechanism, with rates below 35% at CC50 concentrations. Focusing on the THCV-rich extract selected for immunological testing, results showed its selective immunomodulatory effect, with clear differences between eutrophic and obese subjects. Treatment maintained IL-10 and IL-4 levels while reducing TNF-α expression in monocytes and CD4+ T cells from obese individuals, indicating a strong anti-inflammatory profile compared to CBG- and CBD-rich extracts. ELISA analysis showed that THCV reduced IL-17 and IFN-γ secretion in obese subjects. Obese individuals also showed higher frequencies of CD220+ monocytes, with no significant changes in p-RPS6K1 expression. THCV reduced MPO+ monocytes in eutrophic but not obese subjects. Additionally, IL-17A+ CD4+ T cell frequencies

decreased in obese subjects after treatment, along with lower CD220 expression and reduced p- RPS6K1 phosphorylation. The lower frequency of CD4+ CD39+CD73+ regulatory T cells in obese

individuals was not reversed by the treatment. Overall, these findings highlight the crucial role of basal immunometabolic status in determining phytocannabinoid responses, suggesting that THCV can modulate inflammatory, regulatory, and intracellular signaling pathways involved in obesity-related immune dysfunction.