Nitric oxide attenuates the expression of natriuretic peptide receptor C and associated adenylyl cyclase signaling in aortic vascular smooth muscle cells: role of MAPK

Abstract

Arejian M, Li Y, Anand-Srivastava MB. Nitric oxide attenuates the expression of natriuretic peptide receptor C and associated adenylyl cyclase signaling in aortic vascular smooth muscle cells: role of MAPK. Am J Physiol Heart Circ Physiol 296: H1859 -H1867, 2009. First published February 27, 2009 doi:10.1152/ajpheart.01108.2008We have earlier shown that the treatment of A10 vascular smooth muscle cells with S-nitroso-N-acetyl-penicillamine (SNAP); nitric oxide donor (NO) for 24 h decreased the expression of natriuretic peptide receptor C (NPR-C) and adenylyl cyclase signaling. The present study was undertaken to examine the implication of different signaling mechanisms in a NO-induced response. The treatment of A10 vascular smooth muscle cells with SNAP decreased the expression of NPR-C and Gi␣ proteins in a time-dependent manner. The expression of Gi␣ proteins was decreased at 6 h, whereas the expression of NPR-C was attenuated at 2 h. The NPR-C-mediated inhibition of adenylyl cyclase was attenuated (ϳ50%) after 2 h of treatment and was completely abolished after 6 h of treatment. The decreased expression of NPR-C and NPR-C-mediated attenuation of adenylyl cyclase after 2 h of treatment was reversed to control levels by PD-98059, a MEK inhibitor. SNAP also modulated the ERK1/2 phosphorylation in a time-dependent manner; an increase was observed up to 2 h, and, thereafter, the ERK1/2 phosphorylation was decreased. On the other hand, 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one and KT-5823 inhibitor of soluble guanylyl cyclase and protein kinase G, respectively, and Mn(III)tetrakis(4-benzoic acid)porphyrin, a scavenger of peroxynitrite, were unable to restore the SNAPinduced decreased expression of NPR-C protein and increased ERK1/2 phosphorylation to control levels. However, the decreased levels of phosphorylated ERK1/2 and Gi␣ proteins were restored to control levels by 8-bromo-cAMP. These results indicate that a temporal relationship follows between a NO-induced decreased expression of NPR-C and Gi␣ proteins. The decreased expression of NPR-C is mediated through cGMP-independent but MAPK-dependent pathway, whereas NO-induced decreased levels of cAMP may contribute to the decreased activation of MAPK and thereby decreased the expression of Gi␣ proteins. G proteins; mitogen-activated protein kinase NATRIURETIC PEPTIDES (NPs) are a family of three peptide hormones termed atrial NP (ANP), brain NP, and C-type NP, all of which are produced in mammalian hearts, including human hearts (31). ANP regulates a variety of physiological parameters, including blood pressure (7), by interacting with receptors on plasma membrane either to generate second messengers such as cAMP (2, 3) and cGMP (20,48) or to affect ion channels (7). Three types of NP receptors (NPRs) have been cloned; NPR-A (16) and NPR-B (15) subtypes are membranebound guanylate cyclases (130-180 kDa) that mediate the biological actions of NP through a regulation of intracellular cGMP levels (7), whereas NPR-C (4, 17) is a disulfide-bridged homodimer of 67-an...

Nitric oxide attenuates overexpression of Giα proteins in vascular smooth muscle cells from SHR: Role of ROS and ROS-mediated signaling

“…As reported earlier [34, 38], the phosphorylation of ERK1/2 was significantly augmented by about 40% in VSMC from SHR compared with WKY and SNAP treatment attenuated the enhanced phosphorylation of ERK1/2 to control levels. In addition, this treatment also attenuated the ERK1/2 phosphorylation by about 25% in VSMC from WKY rats.…”

“…In addition, NO has been shown to regulate MAPK activity [38, 48, 49]. To investigate the role of MAP kinase in SNAP-induced attenuation of enhanced expression of Giα proteins in VSMC from SHR, we inhibited MAP kinase activity by pretreating the cells with PD98059, a MEK inhibitor and then examined the effect of inhibition of MAP kinase on SNAP- induced decreased expression of Giα protein in VSMC from SHR and WKY rats.…”

“…Subconfluent cells were then starved by incubation for 3 h in DMEM without FBS at 37°C to reduce the interference by growth factors present in the serum. To examine the effect of ODQ or PD98059 on SNAP-induced decreased expression of Giα proteins, the cells were preincubated in the absence (control) or presence of ODQ (20 μM), or PD98059 (10 μM) for 30 mins prior to the treatment with SNAP (100 μM) for 24 h. This time of treatment with SNAP has been shown to decrease the expression of Giα proteins and natriuretic peptide receptor-C in VSMC [37, 38] and suggests that SNAP at 24 h of treatment is stable and active. After incubation, the cells were washed twice with ice-cold phosphate-buffered saline (PBS) and lysed in a 200 μl buffer containing 25 mM Tris·HCl (pH 7.5), 25 mM NaCl, 1 mM sodium orthovanadate, 10 mM sodium fluoride, 10 mM sodium pyrophosphate, 2 mM EDTA, 1 mM phenylmethylsulfonyl fluoride, 10 μg/ml aprotinin, 1% Triton X-100, 0.1% sodium dodecyl sulfate, and 0.5 μg/ml leupeptin on ice.…”

Kinin B1receptor upregulation by angiotensin II and endothelin-1 in rat vascular smooth muscle cells: receptors and mechanisms

“…The cell lysates were centrifuged at 12,000 rpm for 5 min at 4°C, and the supernatants were used for Western blot analysis. Cell viability was checked with the trypan blue exclusion technique and indicated that Ͼ90 ϳ 95% cells were viable, like described previously (4).…”

Modulatory Role of Nitric Oxide/cGMP System in Endothelin-1-Induced Signaling Responses in Vascular Smooth Muscle Cells

“…2A and 2B ) [76]. Since SNAP raises intracellular cGMP levels via sGC in A10 VSMC [65,66], additional studies demonstrated that 8-Br-cGMP (8-bromoguanosine 3’, 5’-cyclic monophosphate), a non-hydro-lyzable analogue of cGMP, mimicked the effect of SNAP and SNP and inhibited ET-1 stimulated ERK1/2 phosphorylation (Fig. 2C ) [76].…”