Research Paper
Subject Category: Cardiovascular and pulmonary pharmacology
British Journal of Pharmacology (2008) 153, 347–357; doi:10.1038/sj.bjp.0707569; published online 12 November 2007
CB2 cannabinoid receptor agonists attenuate TNF-
-induced human vascular smooth muscle cell proliferation and migration
M Rajesh1, P Mukhopadhyay1, G Haskó2, J W Huffman3, K Mackie4 and P Pacher1
- 1Section on Oxidative Stress Tissue Injury, Laboratory of Physiological Studies, National Institutes of Health, NIAAA, Bethesda, MD, USA
- 2Department of Surgery, UMDNJ-New Jersey Medical School, Newark, NJ, USA
- 3Howard L Hunter Chemistry Laboratory, Clemson University, Clemson, SC, USA
- 4Department of Psychological and Brain Sciences, Indiana University, Bloomington, IN, USA
Correspondence: Dr P Pacher, Laboratory of Physiological Studies, National Institutes of Health/NIAAA, 5625 Fishers Lane, MSC-9413, Bethesda, MD 20892-9413, USA. E-mail: pacher@mail.nih.gov
Received 10 October 2007; Accepted 16 October 2007; Published online 12 November 2007.
Abstract
Background and purpose:
Vascular smooth muscle proliferation and migration triggered by inflammatory stimuli are involved in the development and progression of atherosclerosis and restenosis. Cannabinoids may modulate cell proliferation in various cell types through cannabinoid 2 (CB2) receptors. Here, we investigated the effects of CB2 receptor agonists on TNF-
-induced proliferation, migration and signal transduction in human coronary artery smooth muscle cells (HCASMCs).
Experimental approach:
HCASMCs were stimulated with TNF-. Smooth muscle proliferation was determined by the extent of BrdU incorporation and the migration was assayed by modified Boyden chamber. CB2 and/or CB1 receptor expressions were determined by immunofluorescence staining, western blotting, RT-PCR, real-time PCR and flow cytometry.
Key results:
Low levels of CB2 and CB1 receptors were detectable in HCASMCs compared to the high levels of CB2 receptors expressed in THP-1 monocytes. TNF- triggered up to 
80% increase (depending on the method used) in CB2 receptor mRNA and/or protein expression in HCASMCs, and induced Ras, p38 MAPK, ERK 1/2, SAPK/JNK and Akt activation, while increasing proliferation and migration. The CB2 agonists, JWH-133 and HU-308, dose-dependently attenuated these effects of TNF-.
Conclusions and implications:
Since the above-mentioned TNF--induced phenotypic changes are critical in the initiation and progression of atherosclerosis and restenosis, our findings suggest that CB2 agonists may offer a novel approach in the treatment of these pathologies by decreasing vascular smooth muscle proliferation and migration.
Keywords:
cannabinoids, cannabinoid 2 receptor, smooth muscle, proliferation, migration, antibodies
Abbreviations:
AEA, anandamide; 2-AG, 2-arachidonoylglycerol; AM-630, (6-iodo-2-methyl-1-[2-(4-morpholinyl)ethyl]-1H-indol-3-yl](4-methoxyphenyl)methanone; CB1 and CB2 receptors, cannabinoid 1 and 2 receptors; HCASMCs, human coronary artery smooth muscle cells; HU-308, (+)-(1aH,3 H,5aH)-4-[2,6-dimethoxy-4-(1,1-dimethylheptyl) phenyl]-6,6-dimethylbicyclo[3.1.1]hept-2-ene-2-carbinol; JWH-133, 3-(1,1-dimethylbutyl)-1-deoxy-
8-tetrahydrocannabinol; MAPK, mitogen-activated protein kinase, SR2 or SR144528; THC, delta9-tetrahydrocannabinol; THP-1, human monocyte cell line; TNF-, tumour necrosis factor alpha
