Lucas Alvim Santos Romani1, João Paulo Dorta Machado2, André Schwambach Vieira2, Alessandra Sussulini1
Currently, neurological diseases affect one-third of the global population, leading to high morbidity and treatment costs. Cannabidiol (CBD) has emerged as a promising alternative for the treatment of several such disorders, including epilepsy, Parkinson’s disease, Alzheimer’s disease, cerebral ischemia, and depression, due to its anti-inflammatory, anxiolytic, and neuroprotective properties. Since its approval by the U.S. Food and Drug Administration (FDA) in 2018 for the treatment of rare forms of epilepsy, CBD has progressively gained approval and regulation in various countries.
Unlike tetrahydrocannabinol (THC), the compound responsible for the psychoactive effects of Cannabis Sativa, CBD has low affinity for CB1 and CB2 cannabinoid receptors and, therefore, does not induce psychoactive effects. This characteristic is one of the key factors contributing to its safety profile, allowing high doses, up to 1500 mg/day, to be well tolerated in humans without significant adverse effects.
However, adverse effects associated with CBD consumption have still been reported in both humans and animal models. Clinical studies have highlighted side effects such as appetite changes, gastrointestinal discomfort, fatigue, and elevated liver enzyme levels. Additionally, animal studies have shown that high doses of CBD can lead to liver injury, hepatocellular hypertrophy, and changes in liver enzyme activity.
Thus, knowledge about the physiological effects of CBD remains limited, particularly regarding mechanistic insights into hepatic alterations. There is a scarcity of in vitro and in vivo studies focused on liver tissues that could provide this type of information. In this context, strategies such as metabolomics and other omics sciences have emerged as powerful tools, offering broad and insightful information on biological systems.
Therefore, this study aims to investigate the molecular mechanisms of CBD action in liver tissues of animal models using a metabolomics approach based on liquid chromatography coupled to high-resolution mass spectrometry. To achieve this, C57BL/6JUnib mice were treated with different CBD doses (4, 20, and 100 mg/kg). Liver metabolites were extracted using a biphasic method to separate nonpolar from polar compounds, which are respectively analyzed using reversed-phase liquid chromatography (RPLC) and hydrophilic interaction liquid chromatography (HILIC), in order to ensure broad metabolite coverage.
Up to this stage, results from the initial sample preparation and metabolite extraction optimization have been obtained. On average, 6,500 features were identified per sample, with only 15% overlap between the different data acquisition modes (RPLC and HILIC). The next steps include statistical processing and biological interpretation of the data using platforms such as GNPS2, MetaboAnalyst, and R programming. The goal is to identify the main altered metabolites and metabolic pathways to support the discussion and conclusions of this work.
As perspectives, metabolomics data from two brain regions, hippocampus and prefrontal cortex, of the same animals are expected to be obtained, allowing the evaluation of CBD’s effects on the central nervous system. To support the interpretation of these results, additional data will be further acquired using desorption electrospray ionization mass spectrometry imaging (DESI-MSI).
Agradecimentos: The authors acknowledge the financial support from FAPESP, CAPES, and CNPq, as well as the Institute of Chemistry, University of Campinas for providing the infrastructure for this research.