We have previously shown that anandamide and its metabolically stable analog, methanandamide, produces vasorelaxation via the production of nitric oxide in rabbit aortic ring preparations, acting on a Gi-coupled, endothelial non-CB1/CB2 putative “anandamide receptor” (Mukhopadhyay et al., 2002). Recently we have demonstrated that anandamide activates endothelial nitric oxide synthase (eNOS) via the activation of Gi protein and PI3K-Akt pathway. In the additional investigation we have found that anandamide produced angiogenic responses via the activation of Akt pathway in endothelial cells which could not be mimicked by the cannabinoid receptor agonists WIN55212 or CP 55940.
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CB1 receptor antagonist SR141716A or LY320135 failed to block anandamide –mediated angiogenesis suggesting the involvement of anandamide receptor in this response. Within this study we found that methanandamide produced an increase in MMP-2 and MMP-9 activity in CB1 receptor blocked LNCaP prostate cancer cells. In addition MMP-2 and MMP-9 activity was down-regulated in “anandamide receptor” blocked LNCaP cells in the presence of both WIN55212 and methanandamide. These data sets suggest that the molecular mechanism of matrix metalloproteinase activity in cell migration is regulated by the anandamide-mediated activity of the CB1 receptor and the anandamide receptor. Data characterizing the role anandamide-mediated NO production in prostate cancer cells MMP-2 and MMP-9 remains inconclusive.
Marijuana’s psychoactive ingredient is known as -9tetrahydrocannabinol (THC). Studies of THC have uncovered a family of rhodopsin-like G-protein coupled receptors as the target of this compound. Researched conducted in the rat brain and the spleen lead to the identification and cloning of CB1R and CB2R (Matsuda et al., 1990; Gerard et al., 1991). The identification of the cannabinoid receptors fueled the hypothesis that the endogenous ligands to these receptors would bear structural similarities to the THC (Mechoulam et al., 1995). The isolation of the first member this family, now called endogenous cannabinoids or endocannabinoids, arachidonoylethanolamide (anandamide) supported this hypothesis (Devane et al., 1992). Studies into the pharmalogical profile of anandamide suggested an additional an additional binding site for this compound.
The investigations into the differential actions of anandamide in endothelial cells, within our research group, suggested the presence of another G protein-coupled receptor, the putative an anandamide receptor (Mukhopadhyay et al., 2002; McCollum et al., 2007). These data indicated an anandamide receptor mediated vasorelaxation response in rabbit aortic endothelial cells (RAEC) and human umbilical vein endothelial cells (HUVEC) (Mukhopadhyay et al., 2002; McCollum et al., 2007). The findings in our laboratory were supported by similar observations in various another microvascular structures (Kunos et al., 2000a, Kunos et al., 2000b, Kunos 2002). Further experimentation within our laboratory revealed the attenuation of proangiogenic response with si-CB1HUVEC cells and CB1R knockout mice with the administration of O-1918. These data series lend support to the role of anandamide receptor in the initiation of angiogenesis.
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