ABSTRACT
Title
EFFECT OF CannaBiNOIDS in b-Amyloyd-induced toxicityin vitro
Authors
T. Iuvone*, D. De Filippis*, M. Cipriano*, A. Russo°, G. Russo°, G. Esposito§, V. Di Marzo#, L. Steardo§,
*Dep.tment of Experimental Pharmacology, University of Naples Federico II, Naples, Italy §Dep.tment of Physiology and Pharmacology V. Erspamer, University of Rome “La Sapienza” Rome, Italy,° Dip. di Scienze Biomorfologiche, University Federico II, Via Sergio Pansini 5, 80131 Naples, Italy, #Institute of Biomolecular Chemistry, Consiglio Nazionale delle Ricerche, Pozzuoli, Naples.
*Dep.tment of Experimental Pharmacology, University of Naples Federico II, Naples, Italy §Dep.tment of Physiology and Pharmacology V. Erspamer, University of Rome “La Sapienza” Rome, Italy,° Dip. di Scienze Biomorfologiche, University Federico II, Via Sergio Pansini 5, 80131 Naples, Italy, #Institute of Biomolecular Chemistry, Consiglio Nazionale delle Ricerche, Pozzuoli, Naples.
Abstract
Introduction. Investigations aimed at exploring cannabidiol (CBD) effects on b-amyloid (Ab)-induced toxicity demonstrated that this cannabinoid was able to protect differentiated PC12 neuronal cells from the detrimental insult induced by Aβpeptide exposure, through a combination of antioxidant, anti-inflammatory, and anti-apoptotic properties. The beneficial effects of CBD were also confirmed in an “in vivo” model of AD-related neuro-inflammation induced by the intra-hippocampal injection of the human Aβ(1-42) fragment in mice. CBD, dose-dependently, inhibited Aβ-induced reactive gliosis in mice by attenuating glial cell activation and pro-inflammatory mediators release.
The considerable and well-documented neuro-protective, anti-inflammatory, and anti-oxidant properties of CBD encouraged to test other non-THC cannabinoids, such as cannabicromene (CBC), cannabigerol (CBG) and cannabidivarin (CBDV), in Aβ-induced toxicity in vitro.
Materials and Methods. Rat glial cells (C6) were cultured in 10% FBS supplemented DMEM, PC12 rat neuronal cells were cultured in 10% FBS plus 5% horse serum supplemented DMEM and SHY-5SY, human neuroblastoma cells, were cultured in 10% FBS supplemented RPMI. Neuronal cells were differentiated with retinoic acid. After 24 h of starvation C6 cells and differentiated PC12 and SHY-5SY cells were treated with 1 mg/mL of Ab(1-42) in the presence or absence of CBC, CBG and CBDV at concentration of 10-9 to 10-6 M. Cell viability was assessed by MTT assay; nitrite production was measured by Griess reaction. In the same experimental conditions RT-PCR was used to evaluate iNOS, transcription.
Results. The obtained results indicate that both CBC and CBDV, but not CBG, reducedreactive gliosis resulting from treatment of C6 cells with Ab. In fact, CBC and CBDV treatments decreased both the nitrite production and cell proliferation in Ab-stimulated C6 cells. Moreover the administration of CBDV prevented the transcription of iNOS protein.
In parallel experiments with Ab-treated PC12 cells, CBC, CBG and CBDV showed no significant effect on Ab-neurotoxicity. Finally,only CBDV was able to prevent, in a concentration dependent manner, Ab-induced neuronal death in differentiated SHSY-5SY cells.
Conclusions. These data indicate that both CBC and CBDV are able to exert in vitro a significant effect on the control of Ab-induced reactive gliosis and neurotoxicity. Nevertheless, further investigations are necessary to clarify the anti-inflammatory and neuroprotective effects exhibited by these compounds.
This work was supported by GW Pharmaceuticals.
The considerable and well-documented neuro-protective, anti-inflammatory, and anti-oxidant properties of CBD encouraged to test other non-THC cannabinoids, such as cannabicromene (CBC), cannabigerol (CBG) and cannabidivarin (CBDV), in Aβ-induced toxicity in vitro.
Materials and Methods. Rat glial cells (C6) were cultured in 10% FBS supplemented DMEM, PC12 rat neuronal cells were cultured in 10% FBS plus 5% horse serum supplemented DMEM and SHY-5SY, human neuroblastoma cells, were cultured in 10% FBS supplemented RPMI. Neuronal cells were differentiated with retinoic acid. After 24 h of starvation C6 cells and differentiated PC12 and SHY-5SY cells were treated with 1 mg/mL of Ab(1-42) in the presence or absence of CBC, CBG and CBDV at concentration of 10-9 to 10-6 M. Cell viability was assessed by MTT assay; nitrite production was measured by Griess reaction. In the same experimental conditions RT-PCR was used to evaluate iNOS, transcription.
Results. The obtained results indicate that both CBC and CBDV, but not CBG, reducedreactive gliosis resulting from treatment of C6 cells with Ab. In fact, CBC and CBDV treatments decreased both the nitrite production and cell proliferation in Ab-stimulated C6 cells. Moreover the administration of CBDV prevented the transcription of iNOS protein.
In parallel experiments with Ab-treated PC12 cells, CBC, CBG and CBDV showed no significant effect on Ab-neurotoxicity. Finally,only CBDV was able to prevent, in a concentration dependent manner, Ab-induced neuronal death in differentiated SHSY-5SY cells.
Conclusions. These data indicate that both CBC and CBDV are able to exert in vitro a significant effect on the control of Ab-induced reactive gliosis and neurotoxicity. Nevertheless, further investigations are necessary to clarify the anti-inflammatory and neuroprotective effects exhibited by these compounds.
This work was supported by GW Pharmaceuticals.