Related Experiment Video
Updated: Jun 4, 2026
07:13
Platelet-based Detection of Nitric Oxide in Blood by Measuring VASP Phosphorylation
Published on: January 7, 2019
The anandamide effect on NO/cGMP pathway in human platelets
Maria Grazia Signorello1, Enrica Giacobbe, Mario Passalacqua
1Department Experimental Medicine, Biochemistry Section, Genoa University, Genoa, Italy.
Journal of Cellular Biochemistry
|February 18, 2011
Summary
Anandamide, an endocannabinoid, boosts platelet nitric oxide (NO) and cGMP levels by activating the PI3K/AKT pathway. This enhances endothelial nitric oxide synthase (eNOS) activity, ultimately extending platelet survival.
Area of Science:
- Biochemistry
- Pharmacology
- Hematology
Background:
- The endocannabinoid anandamide plays a role in various physiological processes.
- Nitric oxide (NO) and cyclic guanosine monophosphate (cGMP) are critical signaling molecules in platelet function.
- Endothelial nitric oxide synthase (eNOS) is a key enzyme in NO production.
Purpose of the Study:
- To investigate the effect of anandamide on the platelet nitric oxide (NO)/cGMP pathway.
- To elucidate the signaling mechanisms involved in anandamide-mediated effects on platelets.
- To determine the role of the PI3K/AKT pathway and cannabinoid receptor 1 (CB1) in anandamide's action on platelets.
Main Methods:
- Measurement of NO and cGMP levels in human platelets.
- Assay of endothelial nitric oxide synthase (eNOS) activity.
- Western blot analysis for eNOS and AKT phosphorylation.
- Pharmacological inhibition using LY294002 (PI3K inhibitor), MK2206 (AKT inhibitor), and SR141716 (CB1 antagonist).
Main Results:
- Anandamide dose- and time-dependently increased platelet NO and cGMP levels.
- Anandamide stimulated eNOS activity and phosphorylation at Ser1177.
- Anandamide also stimulated AKT phosphorylation at Ser473, indicating PI3K/AKT pathway activation.
- Inhibitors of PI3K, AKT, and the CB1 receptor blocked the effects of anandamide.
Conclusions:
- Low concentrations of anandamide stimulate eNOS activity and increase NO levels in human platelets via PI3K/AKT pathway activation.
- Anandamide exerts its effects through binding to the cannabinoid receptor 1 (CB1).
- Anandamide contributes to extended platelet survival by modulating the NO/cGMP pathway.
More Related Videos
07:15Measurement of Cyclic Guanosine Monophosphate (cGMP) in Solid Tissues using Competitive Enzyme-Linked Immunosorbent Assay (ELISA)
Published on: July 3, 2025
09:13Turbidimetry on Human Washed Platelets: The Effect of the Pannexin1-inhibitor Brilliant Blue FCF on Collagen-induced Aggregation
Published on: April 6, 2017
Related Concept Videos
01:28Nitric Oxide Signaling Pathway
Nitric oxide (NO), an inorganic gas, acts as a potent second messenger in most animal and plant tissues. NO diffuses out of the cells that produce it and enters the neighboring cells to generate a downstream response. NO synthase (NOS) catalyzes NO production by the deamination of the amino acid arginine. There are three isoforms of NOS. Endothelial cells have endothelial NOS (eNOS), nerve and muscle cells have neuronal NOS (nNOS), and macrophages produce inducible NOS (iNOS) upon exposure to...
01:09GPCRs Regulate Adenylyl Cylase Activity
Some GPCRs transmit signals through adenylyl cyclase (AC), a transmembrane enzyme. AC helps synthesize second messenger cyclic adenosine monophosphate (cAMP). AC catalyzes cyclization reaction and converts ATP to cAMP by releasing a pyrophosphate. The pyrophosphate is further hydrolyzed to phosphate by the enzyme pyrophosphatase, which drives cAMP synthesis to completion. However, cAMP is rapidly degraded to 5′ AMP by the enzymes phosphodiesterase (PDE), preventing overstimulation of cells.
Two...
Two...
01:20Antiplatelet Drugs: Prostaglandin Synthesis, P2Y12 and Glycoprotein IIb/IIIa Inhibitors
Antiplatelet drugs emerge as frontline defenders against the insidious threat of thromboembolic diseases, where abnormal clots obstruct vital blood vessels. These drugs stand as bulwarks, inhibiting platelet aggregation and clot formation, thereby mitigating the risk of life-threatening conditions like myocardial infarction, coronary artery disease, and thrombotic strokes.
Prostaglandin synthesis inhibitors, exemplified by the widely known aspirin, wield their power by irreversibly acetylating...
Prostaglandin synthesis inhibitors, exemplified by the widely known aspirin, wield their power by irreversibly acetylating...
01:11IP3/DAG Signaling Pathway
Membrane lipids such as phosphatidylinositol (PI) are precursors for several membrane-bound and soluble second messengers. Specific kinases phosphorylate PI and produce phosphorylated inositol phospholipids. One such inositol phospholipids are the phosphatidylinositol-4,5 bisphosphate [PI(4,5)P2], present in the inner half of the lipid bilayer. Upon ligand binding, GPCR stimulates Gq proteins to turn on phospholipase Cꞵ. Activated phospholipase Cꞵ cleaves PI(4,5)P2 and produces two-second...
01:15Amplifying Signals via Second Messengers
Many receptor binding ligands are hydrophilic; they do not cross the cell membrane but bind to cell-surface receptors. Thus, their message must be relayed by second messengers present in the cell cytoplasm. There are several second messenger pathways, each with its own way of relaying information. For example, the G protein-coupled receptors can activate both phosphoinositol and cyclic AMP (cAMP) second messenger pathways. The phosphoinositol pathway is active when the receptor induces...
01:25cAMP-dependent Protein Kinase Pathways
Cyclic Adenosine Monophosphate (cAMP) is an essential second messenger that activates protein kinase A (PKA) and regulates various biological processes. A single epinephrine molecule binds to GPCR and activates several heterotrimeric G proteins, each stimulating multiple adenylyl cyclase, amplifying the signal, and synthesizing large numbers of cAMP molecules. Small changes in cAMP concentration affect PKA activity. The binding of four cAMP molecules induces a conformational change in PKA,...