Inhibition of protein tyrosine phosphatases unmasks vasoconstriction and potentiates calcium signaling in rat aorta smooth muscle cells in response to an agonist of 5-HT2B receptors BW723C86

• Published in: Biochemical and biophysical research communications Vol. 483; no. 1; pp. 700 - 705
• Main Authors: Mironova, Galina Y., Avdonin, Piotr P., Goncharov, Nikolay V., Jenkins, Richard O., Avdonin, Pavel V.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 29.01.2017
• Subjects: 5-HT2B receptors; Animals; Aorta - drug effects; Aorta - enzymology; Aorta - physiology; Calcium; Calcium Signaling - drug effects; Cell Separation; Hypertension; Indoles - pharmacology; Male; Mesenteric Arteries - drug effects; Mesenteric Arteries - physiology; Muscle Contraction - drug effects; Muscle, Smooth, Vascular - cytology; Muscle, Smooth, Vascular - drug effects; Muscle, Smooth, Vascular - enzymology; Oxidative Stress; Protein tyrosine phosphatases; Protein Tyrosine Phosphatases - analysis; Rats; Rats, Wistar; Receptor, Serotonin, 5-HT2B - genetics; Receptor, Serotonin, 5-HT2B - metabolism; RNA, Messenger - analysis; Serotonin 5-HT2 Receptor Agonists - pharmacology; Smooth muscle cells; Thiophenes - pharmacology; Vanadates - pharmacology; Vasoconstriction - drug effects; Vasoconstriction - physiology; Vasoconstrictor Agents - pharmacology; Hypertension; Oxidative stress; Protein tyrosine phosphatases; Calcium; Smooth muscle cells; 5-HT2B receptors
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2016.12.079

In blood vessels, serotonin 5-HT2B receptors mainly mediate relaxation, although their activation by the selective agonist BW723C86 is known to exert contraction of aorta in deoxycorticosterone acetate (DOCA)-salt and N(omega)-nitro-l-arginine (l-NAME) hypertensive rats [Russel et al., 2002; Banes et al., 2003] and in mice with type 2 diabetes [Nelson et al., 2012]. The unmasking effect on vasoconstriction can be caused by a shift in the balance of tyrosine phosphorylation in smooth muscle cells (SMC) due to oxidative stress induced inhibition of protein tyrosine phosphatases (PTP). We have demonstrated that BW723C86 which does not cause contraction of rat aorta and mesenteric artery rings, evoked a vasoconstrictor effect in the presence of PTP inhibitors sodium orthovanadate (Na3VO4) or BVT948. BW723C86 induced a weak rise of [Ca2+]i in the SMC isolated from rat aorta; however, after pre-incubation with Na3VO4 the response to BW723C86 increased more than 5-fold. This effect was diminished by protein tyrosine kinase (PTK) inhibitor genistein, inhibitor of Src-family kinases PP2, inhibitor of NADPH-oxidase VAS2870 and completely suppressed by N-acetylcysteine and 5-HT2B receptor antagonist RS127445. Using fluorescent probe DCFH-DA we have shown that Na3VO4 induces oxidative stress in SMC. In the presence of Na3VO4 BW723C86 considerably increased formation of reactive oxygen species while alone had no appreciable effect on DCFH oxidation. We suggest that oxidative stress causes inhibition of PTP and unmasking of 5-HT2B receptors functional activity.