Cannabidiol modulates NETs formation by neutrophils stimulated by amyloid fibers, Leishmania amazonensis, and PMA.

Cannabidiol (CBD) is known for its anti-inflammatory and antioxidant properties1. Neutrophils play a crucial role in immune functions by releasing neutrophil extracellular traps (NETs), composed of extracellular decondensed DNA and proteins that capture and neutralize pathogens. The formation of NETs can be triggered by various stimuli, including microorganisms and amyloid fibers, and have been implicated in autoimmune disorders by promoting inflammation2. It has been shown that CBD reduces NET formation in neutrophils from patients with psoriasis, although CBD alone stimulates the formation of NETs3. Here, we investigate the role of CBD and Delta-9-tetrahydrocannabinol (THC)-CBD (1:1) in modulating NET formation and ROS production by various stimuli. Thus, human neutrophils were pretreated with CBD or THC-CBD for 30 minutes and then stimulated with L. amazonensis, α-synuclein, or phorbol 12-myristate 13-acetate (PMA) for 4h at 37ºC. NETs were quantified by the DNA marker Picogreen and reactive oxygen species (ROS) by the probe dihydrorhodamine (DHR)-123. Our results indicate that CBD or THC-CBD at the concentrations tested did not induce NETs or were toxic for the neutrophils measured by lactate dehydrogenase release. However, pretreatment with CBD enhances NET release upon stimulation with L. amazonensis and α-synuclein through a mechanism that does not involve ROS. Conversely, CBD inhibits NET formation stimulated with PMA. ROS production was reduced by CBD after stimulation with PMA, and α-syn and THC-CDB, only for PMA. In conclusion, we show that CBD and THC-CBD modulate NET and ROS production in a stimulus-dependent way. 1- Antioxidants. (2019) 9(1), 21. 2- Nat Rev Immunol (2023) 23, 274–288. 3- nt J Mol Sci. (2020) 16;21(18):6795