title: Cytotoxicity of synthetic cannabinoids on primary neuronal cells of the forebrain: the involvement of cannabinoid CB1 receptors and apoptotic cell death.
abstract: The abuse of herbal products containing synthetic cannabinoids has become an issue of public concern. The purpose of this paper was to evaluate the acute cytotoxicity of synthetic cannabinoids on mouse brain neuronal cells. Cytotoxicity induced by synthetic cannabinoid (CP-55,940, CP-47,497, CP-47,497-C8, HU-210, JWH-018, JWH-210, AM-2201, and MAM-2201) was examined using forebrain neuronal cultures. These synthetic cannabinoids induced cytotoxicity in the forebrain cultures in a concentration-dependent manner. The cytotoxicity was suppressed by preincubation with the selective CB1 receptor antagonist AM251, but not with the selective CB2 receptor antagonist AM630. Furthermore, annexin-V-positive cells were found among the treated forebrain cells. Synthetic cannabinoid treatment induced the activation of caspase-3, and preincubation with a caspase-3 inhibitor significantly suppressed the cytotoxicity. These synthetic cannabinoids induced apoptosis through a caspase-3-dependent mechanism in the forebrain cultures. Our results indicate that the cytotoxicity of synthetic cannabinoids towards primary neuronal cells is mediated by the CB1 receptor, but not by the CB2 receptor, and further suggest that caspase cascades may play an important role in the apoptosis induced by these synthetic cannabinoids. In conclusion, excessive synthetic cannabinoid abuse may present a serious acute health concern due to neuronal damage or deficits in the brain.
authors.affiliation: Department of Drug Dependence Research, National Institute of Mental Health, National Center of Neurology and Psychiatry, Kodaira, Tokyo, Japan.
authors.forename: Ken-ichi
authors.initials: K
authors.lastname: Tomiyama
authors.affiliation: Department of Drug Dependence Research, National Institute of Mental Health, National Center of Neurology and Psychiatry, Kodaira, Tokyo, Japan. Electronic address: mfunada@ncnp.go.jp.
authors.forename: Masahiko
authors.initials: M
authors.lastname: Funada
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D002186:Cannabinoids / Q000138:chemical synthesis / Q000633:toxicity*; D002478:Cells, Cultured; D003603:Cytotoxins / Q000138:chemical synthesis / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D005260:Female; D051379:Mice; D008813:Mice, Inbred ICR; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D011247:Pregnancy; D016548:Prosencephalon / Q000187:drug effects* / Q000378:metabolism; D043884:Receptor, Cannabinoid, CB1 / Q000096:biosynthesis*
chemical_list: D002186:Cannabinoids; D003603:Cytotoxins; D043884:Receptor, Cannabinoid, CB1
keywords: Apoptosis; CB; Cannabinoid receptor; Cytotoxicity; DMEM; DMSO; Designer drug; Dulbecco's modified Eagle's medium; FBS; MAP-2; PBS; Primary neuronal cells; RT; Synthetic cannabinoid; Z-Asp(OMe)-Glu(OMe)-Val-Asp(OMe)-fluoromethylketone; Z-DEVD-FMK; cannabinoid; dimethyl sulfoxide; fetal bovine serum; microtubule-associated protein-2; phosphate-buffered saline; room temperature; Δ(9)-THC; ∆(9)-tetrahydrocannabinol