Effects of physiological and supraphysiological concentrations of cannabidiol on pre‐adipocyte proliferation

Abstract

Cannabidiol (CBD) is one of the non‐psychotropic endocannabinoids from the Cannabis sativa plant. Studies support that CBD can modulate inflammation and lipid production in various cell lines, but its anti‐obesity effects are not well defined. Specifically, the effects of CBD on differentiation and proliferation of adipocytes are still unknown. Excessive fat accumulation can result in obesity, a worldwide public health problem with an increasing trend. Obesity increases the risk of developing chronic diseases. The purpose of this study was to determine the effects of different concentrations of CBD (0.2 [physiological] and 20 μM [supraphysiological]) on proliferation of pre‐adipocytes (3T3‐L1 cell line). Cells were cultured at 37°C in a humidified atmosphere (5% CO2) until they became pre‐confluent. Three groups of cells were set up: control (cells in complete growth media [CGM]), CGM supplemented with 0.2 μM CBD, and CGM supplemented with 20 μM CBD. Cells were plated in 96‐well plates in octuplicates in media containing the corresponding treatment (C, 0.2 μM CBD, or 20 μM CBD). Cells were incubated for 6, 12, 24 and 48 h, at which time points cell proliferation was measured using an MTS assay. At the 24 and 48 hour time points, cells treated with 20 μM CBD had higher cell proliferation (24 hours: 0.299 ± 0.006; 48 hours: 0.449±0.016) compared to the 0.2 μM group (24 hours: 0.262± 0.006; 48 hours: 0.217± 0.016) and the control group (24 hours: 0.249± 0.006; 48 hours: 0.312± 0.016) (p<0.0001, full‐factorial ANOVA). Interestingly, cells treated with 0.2 μM CBD had decreased proliferation at 48 hours compared to control cells, while there was no difference in proliferation between these two groups at 24 hours. Our results suggest that physiological and supraphysiological concentrations of CBD affect pre‐adipocyte proliferation differently and that further research is needed to clarify these effects.