Establishment Of Cell Lines Stably Expressing CB1 Cannabinoid Receptor Or CB2 Cannabinoid Receptor

Cannabinoid receptor 1(CB1)and Cannabinoid receptor 2(CB2)belong to the G protein-coupled receptors(GPCRs).CB1 receptor is located mainly in the brain and other nervous tissues.The CB2 receptor is predominantly found in the immune system.Activation of cannabinoid receptors(CB1 and CB2)could lead to the inhibition of adenylyl cyclase and reduct accumulation of cyclic AMP.In addition,both the CB1 and CB2 cannabinoid receptors can regulate the phosphorylation and activate different members of mitogenactivated protein kinases(MAPKs),and finally modulate nociceptive thresholds and inhibit release of pro-inflammatory molecules.It also displays synergistic effects with other systems especially the endogenous opioid system that influence analgesia.Therefore,agonists cannabinoid receptor have shown therapeutic value against neuropathic pains and inflammatory.In order to research activity of endogenous cannabinoid ligands and receptors on the cellular level,we try to establish cells which could stably express CB1 receptor or CB2 receptor.First,CHO-K1 and HEK293A cells were transfected with the plasmid pcDNA3.1-HA-CB1 and pcDNA3.1-HA-CB2 by using Lipofectamine 2000.Then,we selected the positive clones which could stably express receptor,and the positive clones were confirmed by RT-PCR,Western blotting and cAMP.The two cell models can be used to the study of mechanism of endogenous cannabinoid ligands analgesic pharmacology and cell signaling mediated by CB1 receptor or CB2 receptor.