Presence Of Suramin Research Articles

​Effect of Ferula Elaeochytris Root Extract on Smooth Muscle Contraction of Prostate Gland in Rat

Background: Smooth muscle contraction and enlargement of the prostate are important targets for the treatment of lower urinary tract symptoms in patients with benign prostatic hyperplasia. The effect of Ferula elaeochytris (FE) root extract on tissues that play a role on fertility such as the prostate has not been demonstrated yet. The aim of this study is to investigate the effects of FE extract on isolated rat prostate tissue induced by electrical field stimulation (EFS) in vitro. Methods: In this experimental study, 48 male rats were randomly divided into 6 groups (n = 8). Groups were consisting FE (20 µl), FE + Adenosine triphosphate (ATP; 50 μM), FE + prazosin (0.3 μM), FE + Calcium (Ca2+; 3 mM and 6 mM), FE + suramin (100 μM), FE + phenylephrine and FE + carbachol. Result: In our study, FE extract inhibited the neurogenic contractions induced by EFS on isolated rat anterior prostate tissue. The presence of suramin and prazosin were not significantly increase the inhibition caused by the FE extract, while Ca2+ and ATP significantly increased the inhibition by the FE extract. (p less than 0.05). In addition, FE extract significantly inhibited phenylephrine and carbachol contractions (p less than 0.05).

2-Cl-C.OXT-A stimulates contraction through the suppression of phosphodiesterase activity in human induced pluripotent stem cell-derived cardiac organoids.

Background2-Cl-C.OXT-A (COA-Cl) is a novel synthesized adenosine analog that activates Sphingosine-1-phosphate 1 receptor (S1P1R) and combines with the adenosine A1 receptor (A1R) in G proteins and was shown to enhance angiogenesis and improve the brain function in rat stroke models. However, the role of COA-Cl in hearts remains unclear. COA-Cl, which has a similar structure to xanthine derivatives, has the potential to suppress phosphodiesterase (PDE), which is an important factor involved in the beating of heart muscle.Methods and resultsCardiac organoids with fibroblasts, human induced pluripotent stem cell-derived cardiac myocytes (hiPSC-CMs), and hiPSC-derived endothelial cells (hiPSC-ECs) were cultured until they started beating. The beating and contraction of organoids were observed before and after the application of COA-Cl. COA-Cl significantly increased the beating rate and fractional area change in organoids. To elucidate the mechanism underlying these effects of COA-Cl on cardiac myocytes, pure hiPSC-CM spheroids were evaluated in the presence/absence of Suramin (antagonist of A1R). The effects of COA-Cl, SEW2871 (direct stimulator of S1P1R), two positive inotropes (Isoproterenol [ISO] and Forskolin [FSK]), and negative inotrope (Propranolol [PRP]) on spheroids were assessed based on the beating rates and cAMP levels. COA-Cl stimulated the beating rates about 1.5-fold compared with ISO and FSK, while PRP suppressed the beating rate. However, no marked changes were observed with SEW2871. COA-Cl, ISO, and FSK increased the cAMP level. In contrast, the level of cAMP did not change with PRP or SEW2871 treatment. The results were the same in the presence of Suramin as absence. Furthermore, an enzyme analysis showed that COA-Cl suppressed the PDE activity by half.ConclusionsCOA-Cl, which has neovascularization effects, suppressed PDE and increased the contraction of cardiac organoids, independent of S1P1R and A1R. These findings suggest that COA-Cl may be useful as an inotropic agent for promoting angiogenesis in the future.

Rat aorta relaxation induced by the venom of Poecilotheria regalis involves the activation of the NO/cGMP pathway

Spider venoms are widely recognized as a new emerging source of potential research tools, pesticides, drug leads, and therapeutic agents. Some studies suggest that these venoms may contain interesting vasodilator compounds with potential therapeutic applications. In the present study, the vasodilator activity of the venom of Poecilotheria regalis was evaluated in isolated rat aortic rings. This venom induced an endothelium-dependent vasodilation [EC50 value was 5.52 (4.18–7.32) μg protein/ml with an Emax = 103.4 ± 3.8%]. While the percentage of vasodilation induced by the venom was significantly diminished in the presence of a nitric oxide synthase inhibitor (L-NAME), it remained unaltered in the presence of suramin, a P2-purinergic receptor antagonist. Moreover, the vasodilator activity of the venom was not affected after boiling bath incubation, but was significantly decreased under reducing conditions. Additionally, venom composition was analyzed by reverse-phase chromatography and MALDI-TOF mass spectrometry, and two fractions were obtained, referred to as peptidic and non-peptidic fractions. Interestingly, both fractions induced vasodilation in isolated rat aortic rings. The results of this study showed that the venom of P. regalis induces a concentration-dependent vasodilation in rat aorta that was endothelium-dependent and involves the activation of NO/cGMP pathway. These results suggest that the venom contains a combination of both peptidic and non-peptidic vasodilator components. This study provides pharmacological data that suggest that P. regalis venom may be an important source of peptidic and non-peptidic vasodilator compounds.

Modulation of dopamine-mediated facilitation at the neuromuscular junction of Wistar rats: A role for adenosine A1/A2A receptors and P2 purinoceptors

This study aims to understand how dopamine and the neuromodulators, adenosine and adenosine triphosphate (ATP) modulate neuromuscular transmission. Adenosine and ATP are well-recognized for their regulatory effects on dopamine in the central nervous system. However, if similar interactions occur at the neuromuscular junction is unknown. We hypothesize that the activation of adenosine A1/A2A and/or P2 purinoceptors may influence the action of dopamine on neuromuscular transmission. Using the rat phrenic nerve hemi-diaphragm, we assessed the influence of dopamine, adenosine and ATP on the height of nerve-evoked muscle twitches. We investigated how the selective blockade of adenosine A1 receptors (2.5nM DPCPX), adenosine A2A receptors (50nM CSC) and P2 purinoceptors (100μM suramin) modified the effects of dopamine. Dopamine alone increased indirect muscle contractions while adenosine and ATP either enhanced or depressed nerve-evoked muscle twitches in a concentration-dependent manner. The facilitatory effects of 256μM dopamine were significantly reduced to 29.62±2.79% or 53.69±5.45% in the presence of DPCPX or CSC, respectively, relative to 70.03±1.57% with dopamine alone. Alternatively, the action of 256μM dopamine was potentiated from 70.03±1.57, in the absence of suramin, to 86.83±4.36%, in the presence of suramin. It can be concluded that the activation of adenosine A1 and A2A receptors and P2 purinoceptors potentially play a central role in the regulation of dopamine effects at the neuromuscular junction. Clinically this study offers new insights for the indirect manipulation of neuromuscular transmission for the treatment of disorders characterized by motor dysfunction.

Suramin Inhibits Chikungunya Virus Entry and Transmission.

The mosquito-borne Chikungunya virus (CHIKV) is a profound global threat due to its high rate of contagion and the lack of vaccine or effective treatment. Suramin is a symmetric polyanionic naphthylurea that is widely used in the clinical treatment of parasite infections. Numerous studies have reported the broad antiviral activities of suramin; however, inhibition effects against CHIKV have not yet been demonstrated. The aim of this study was thus to investigate the antiviral effect of suramin on CHIKV infection and to elucidate the molecular mechanism underlying inhibition using plaque reduction assay, RT-qPCR, western blot analysis, and plaque assay. Microneutralization assay was used to determine the EC50 of suramin in the CHIKV-S27 strain as well as in three other clinical strains (0611aTw, 0810bTw and 0706aTw). Time-of-addition was used to reveal the anti-CHIKV mechanism of suramin. We also evaluated anti-CHIKV activity with regard to viral entry, virus release, and cell-to-cell transmission. Cytopathic effect, viral RNA, viral protein, and the virus yield of CHIKV infection were shown to diminish in the presence of suramin in a dose-dependent manner. Suramin was also shown the inhibitory activities of the three clinical isolates. Suramin inhibited the early progression of CHIKV infection, due perhaps to interference with virus fusion and binding, which subsequently prevented viral entry. Results of a molecular docking simulation indicate that suramin may embed within the cavity of the E1/E2 heterodimer to interfere with their function. Suramin was also shown to reduce viral release and cell-to-cell transmission of CHIKV. In conclusion, Suramin shows considerable potential as a novel anti-CHIKV agent targeting viral entry, extracellular transmission, and cell-to-cell transmission.

Cell-to-cell propagation of intracellular signals fluorescently visualized with acridine orange in the gastric glands of guinea pigs

Secretion from the gastric gland involves the activation of various types of cells in a coordinated manner. In order to elucidate the mechanisms underlying the coordination of secretion, we studied live fluorescence images of guinea pig gastric glands stained with acridine orange (AO). On 2μM AO staining, individual cells were characterized by metachromatic colors and various intensities of fluorescence. When the gland was stimulated with 100μM of histamine, green fluorescence was transiently increased in parietal cells and intermediate cells and propagated along the gland for a long distance over many cells. Local stimulation in a couple of cells with histamine in the presence of suramin also induced propagation. However, the fluorescence response was suppressed by the addition of H-89, a protein kinase A inhibitor. These findings suggest that a cAMP-dependent signal propagates intercellularly through a variety of cells to induce coordinated secretion in the entire gastric gland.

Mucosal adenosine triphosphate mediates serotonin release from ileal but not colonic guinea pig enterochromaffin cells

Mechanical stimulation of the mucosal epithelium results in increased serotonin (5-HT) release from enterochromaffin (EC) cells. Little is known about how this process varies in different regions of the intestinal tract; however, purines are felt to play a role. We studied the relationship between mechanical stimulation, adenosine triphosphate (ATP), and 5-HT release from ileal and colonic mucosal tissue. Amperometric recordings of ATP and 5-HT were carried out using an ATP biosensor and boron-doped diamond microelectrode. Levels of extracellular ATP and 5-HT were monitored using high performance liquid chromatography. Under basal conditions, 5-HT levels were significantly decreased in the ileum (p<0.001) but not the colon in the presence of the P2 antagonist suramin (100μM). Ecto-ATPase inhibitor ARL67156 (10μM) elevated ATP levels in the ileum and colon (both p<0.001), but only 5-HT levels in the ileum (p<0.001). Exogenous ATP increased 5-HT release in the presence of tetrodotoxin in the ileum (p<0.001), but had not effect in the colon. Mechanical stimulation increased levels of 5-HT in the ileum (p<0.001) and colon (p<0.01), but levels returned to baseline in the presence of suramin and MRS2179 in the ileum. The onset of 5-HT release was delayed following mechanical stimulation. The rise time of the ATP response was quicker than that of 5-HT during mechanical stimulation. During mechanical stimulation of the mucosal epithelium, ATP mediates 5-HT release from EC cells in the ileum, but not the colon. Mucosal 5-HT signaling following mechanical stimulation is varied in different regions of the intestinal tract.

Inhibition of Presynaptic Neurotoxins in Taipan Venom by Suramin

Taipans are amongst the most venomous snakes in the world, and neurotoxicity is a major life-threatening symptom of envenoming by these snakes. Three species of taipans exist, and the venom from each species contains a presynaptic neurotoxin which accounts for much of the neurotoxicity observed following human envenoming. The high cost of antivenom used to treat neurotoxicity has resulted in the need to develop alternative but effective therapies. Therefore, in this study, we examined the ability of the P2Y receptor antagonist suramin to prevent the in vitro neurotoxic effects of the three presynaptic neurotoxins in taipan venoms: taipoxin, paradoxin and cannitoxin. Toxins were purchased from commercial sources or purified in house, using multiple steps of gel filtration chromatography. All three toxins (11 nM) inhibited nerve-mediated twitches in the chick biventer cervicis nerve-muscle preparation within 300 min. The presence of suramin (0.3 mM) completely blocked the taipoxin and cannitoxin-mediated inhibition of nerve-mediated twitches within the course of the experiment (P < 0.0001). However, paradoxin induced a 32 % decrease in twitch height even in the presence of suramin within 360 min. This was significantly different compared to toxin alone (P < 0.0001). We also examined the effect of suramin on the neurotoxic effects of textilotoxin and the products of phospholipase A2 action. Each toxin alone or in the presence of suramin failed to inhibit the responses to exogenous agonists ACh, CCh or KCl. Our results warrant clinical studies aimed determining the efficacy of suramin in preventing the onset of neurotoxicity following taipan envenoming.

Identification of a Potent Endothelium-Derived Angiogenic Factor

The secretion of angiogenic factors by vascular endothelial cells is one of the key mechanisms of angiogenesis. Here we report on the isolation of a new potent angiogenic factor, diuridine tetraphosphate (Up4U) from the secretome of human endothelial cells. The angiogenic effect of the endothelial secretome was partially reduced after incubation with alkaline phosphatase and abolished in the presence of suramin. In one fraction, purified to homogeneity by reversed phase and affinity chromatography, Up4U was identified by MALDI-LIFT-fragment-mass-spectrometry, enzymatic cleavage analysis and retention-time comparison. Beside a strong angiogenic effect on the yolk sac membrane and the developing rat embryo itself, Up4U increased the proliferation rate of endothelial cells and, in the presence of PDGF, of vascular smooth muscle cells. Up4U stimulated the migration rate of endothelial cells via P2Y2-receptors, increased the ability of endothelial cells to form capillary-like tubes and acts as a potent inducer of sprouting angiogenesis originating from gel-embedded EC spheroids. Endothelial cells released Up4U after stimulation with shear stress. Mean total plasma Up4U concentrations of healthy subjects (N = 6) were sufficient to induce angiogenic and proliferative effects (1.34±0.26 nmol L-1). In conclusion, Up4U is a novel strong human endothelium-derived angiogenic factor.

Involvement of a putative intercellular signal-recognizing G protein-coupled receptor in the engulfment of Salmonella by the protozoan Tetrahymena

In an effort to investigate the molecular basis of protozoa engulfment-mediated hypervirulence of Salmonella in cattle, we evaluated protozoan G protein-coupled receptors (GPCRs) as transducers of Salmonella engulfment by the model protozoan Tetrahymena. Our laboratory previously demonstrated that non-pathogenic protozoa (including Tetrahymena) engulf Salmonella and then exacerbate its virulence in cattle, but the mechanistic details of the phenomenon are not fully understood. GPCRs were investigated since these receptors facilitate phagocytosis of particulates by Tetrahymena, and a GPCR apparently modulates bacterial engulfment for the pathogenic protozoan Entamoeba histolytica. A database search identified three putative Tetrahymena GPCRs, based on sequence homologies and predicted transmembrane domains, that were the focus of this study. Salmonella engulfment by Tetrahymena was assessed in the presence of suramin, a non-specific GPCR inhibitor. Salmonella engulfment was also assessed in Tetrahymena in which expression of putative GPCRs was knocked-down using RNAi. A candidate GPCR was then expressed in a heterologous yeast expression system for further characterization. Our results revealed that Tetrahymena were less efficient at engulfing Salmonella in the presence of suramin. Engulfment was reduced concordantly with a reduction in the density of protozoa. RNAi-based studies revealed that knock-down of one the Tetrahymena GPCRs caused diminished engulfment of Salmonella. Tetrahymena lysates activated this receptor in the heterologous expression system. These data demonstrate that the Tetrahymena receptor is a putative GPCR that facilitates bacterial engulfment by Tetrahymena. Activation of the putative GPCR seemed to be related to protozoan cell density, suggesting that its cognate ligand is an intercellular signaling molecule.

Purinergic receptors and regulatory volume decrease in seabream (Sparus aurata) hepatocytes: a videometric study

The response of isolated hepatocytes of Sparus aurata to hypotonic stress was studied by the aid of videometric methods with the aim to investigate the possible involvement of ATP in the regulatory volume decrease (RVD). This study confirms our previous observations showing the ability of these cells to undergo RVD. In addition, it shows that the homeostatic response was inhibited by apyrase, an ATP scavenger, thus suggesting the involvement of extracellular ATP in the RVD response. Experiments performed in the presence of ATPγS or adenosine, agonists of P(2) and P(1) receptors respectively, and in the presence of suramin or 8-PT, antagonists of P(2) and P(1) receptors respectively, suggest that ATP exerts its stimulatory effect on the homeostatic response by interacting with P(2) receptors. On the other hand, the activation of P(1) receptors by ATP metabolites produces opposite effects. In an attempt to clarify the mechanisms involved in ATP release from the cell, we performed some experiments with known inhibitors of the possible mechanisms of regulated ATP release. The results we obtained let us to suppose that the mechanism allowing the exit of ATP from the cell is verapamil sensitive suggesting the involvement of the P-glycoprotein.

Pharmacokinetics, metabolism and excretion of megazol, a new potent trypanocidal drug in animals.

The pharmacokinetics of megazol (CAS 19622-55-0) was investigated after intraperitoneal and oral administration of the drug (80 mg/kg) to mice. The plasma levels were significantly higher after oral administration of drug than after intraperitoneal route (33.8 micrograms/ml compared with 19.0 micrograms/ml for Cmax, 158714 micrograms.h/l compared with 96057 micrograms.h/l for AUC). When suramin (CAS 145-63-1) was administered 24 h before oral administration of megazol, megazol absorption was accelerated (2 h compared with 4 h for Tmax) but the amount absorbed was lower (19.9 micrograms/ml compared with 33.8 micrograms/ml for Cmax and 95547 micrograms.h/l vs 158714 micrograms.h/l for AUC). In the infected mice previously treated with suramin, all estimated pharmacokinetic parameters of plasma megazol were significantly modified, in particularly an increase in the apparent volume of distribution (5.6 l/kg compared with 0.9 l/kg) with a prolongation of the elimination half-life (3 h compared with 0.7 h) of megazol. Excretion of the total radioactivity of megazol was also evaluated after oral administration of 3H-megazol to rats. Total radioactivity was eliminated predominantly via the urinary route (80%) vs. 10.5% in the faeces, 9.5% remaining in the body 8 days after dosing. When unlabelled megazol was orally administered to rats with absence or presence of suramin, megazol recovered in urine and faeces 72 h dosing was: 55.7%/2% vs 20.6%/1.6%, respectively. In the urine, unchanged megazol was present as characterized by LC-MS/MS as well as 4 unknown metabolites. This study indicates that suramin significantly affects the pharmacokinetics of megazol and its elimination.

A Survey of Antiprion Compounds Reveals the Prevalence of Non-PrP Molecular Targets

Prion diseases are fatal neurodegenerative diseases caused by the accumulation of the misfolded isoform (PrP(Sc)) of the prion protein (PrP(C)). Cell-based screens have identified several compounds that induce a reduction in PrP(Sc) levels in infected cultured cells. However, the molecular targets of most antiprion compounds remain unknown. We undertook a large-scale, unbiased, cell-based screen for antiprion compounds and then investigated whether a representative subset of the active molecules had measurable affinity for PrP, increased the susceptibility of PrP(Sc) to proteolysis, or altered the cellular localization or expression level of PrP(C). None of the antiprion compounds showed in vitro affinity for PrP or had the ability to disaggregate PrP(Sc) in infected brain homogenates. These observations suggest that most antiprion compounds identified in cell-based screens deploy their activity via non-PrP targets in the cell. Our findings indicate that in comparison to PrP conformers themselves, proteins that play auxiliary roles in prion propagation may be more effective targets for future drug discovery efforts.

유충 Manduca sexta 리포포린에 의한 지방체로의 디아실글리세리드 운반

This paper was to characterize the transfer of diacylglycerol(DAG) from lipophorin to Manduca sexta larval fat bodies. -DAG-labeled Lp(-DAG-Lp) was incubated with the larval fat bodies under different times and the time of DAG transfer was determined. Incubation of fat bodies with -DAG-Lp resulted in accumulation of DAG and TAG in the tissue. The transfer of -DAG was inhibited in the presence of suramin and unlabeled lipophorin, which would be consistent with a lipophorin receptor. The effects of suramin may be complex because it can change membrane properties when bound to the lipophorin receptor and affect the rate of DAG transfer. To investigate the lipid uptake via receptor-mediated endocytosis, we treated with endocytosis inhibitors, ammonium chloride and chloroquine. The results show that the transfer process of lipid by lipophorin and fat bodies is receptor-mediated endocytosis.

The disposition of pyrimethamine in the rat isolated perfused liver: the effect of suramin.

The pharmacokinetics of pyrimethamine (0.5 mg) were determined in the rat isolated perfused liver in the presence of suramin (17.75 mg). The clearance, half life, area under the curve, and volume of distribution of pyrimethamine were unaffected by the concurrent administration of suramin, as was the hepatic sub-cellular disposition of the drug. This report is of relevance to the future concurrent administration of suramin and pyrimethamine in West Africa.

Contractile responses of chicken rectum to stimulation of Remak's nerve or purinoceptors: effect of suramin.

Stimulation of Remak's nerve produced a rapid contraction of the rectal muscle which was resistant to blockade by hyoscine, followed by a slow contractile response which was cholinergic. With hyoscine (1 microM) in the Tyrode solution, the non-cholinergic contractile response to nerve stimulation was compared with the responses to adenosine triphosphate (ATP) and alpha,beta-methylene adenosine triphosphate (alpha,beta-Me ATP). In the presence of suramin (300 microM), the contractile response to ATP was increased by about 100%, whereas that to nerve stimulation was inhibited by approximately 17%. Suramin (60 microM) also discriminated between the contractile responses to ATP and nerve stimulation, only the former being potentiated by the drug. It seems probable that suramin potentiated the action of ATP in the rectum through inhibition of ectonucleotidase activity. If so, the potentiation should not extend to alpha,beta-Me ATP, as this analogue of ATP is resistant to inactivation by the enzyme. Suramin inhibited the contractile response to alpha,beta-Me ATP. Thus the neurotransmitter which mediated the hyoscine-resistant contractile response to stimulation of Remak's nerve was dissimilar to ATP, in that it was not potentiated by suramin and presumably was not inactivated by ectonucleotidase activity.

In vitro interaction between suramin and fluvastatin against clinically important Zygomycetes

The in vitro antifungal activity of suramin and its combinations with fluvastatin was investigated in this study. Several species belonging to the class Zygomycetes are considered agents of opportunistic human or animal infections. In the present work, 15 fungal isolates representing eight clinically important genera (Absidia, Micromucor, Mortierella, Mucor, Rhizomucor, Rhizopus, Saksenaea and Syncephalastrum) were investigated. The efficiency with which fluvastatin inhibited growth in the presence of suramin (100 microg ml(-1)) was studied. The investigated compounds acted synergistically and additively on the growth when a strain was resistant to suramin and sensitive to fluvastatin; at the same time, antagonistic interactions were detected when strains were sensitive to both agents. In these cases, the growth inhibition effect of suramin was dominant.

Endogenous ATP potentiates only vasopressin secretion from neurohypophysial terminals

Exogenous ATP induces inward currents and causes the release of arginine-vasopressin (AVP) from isolated neurohypophysial terminals (NHT); both effects are inhibited by the P2X2 and P2X3 antagonists, suramin and PPADS. Here we examined the role of endogenous ATP in the neurohypophysis. Stimulation of NHT caused the release of both AVP and ATP. ATP induced a potentiation in the stimulated release of AVP, but not of oxytocin (OT), which was blocked by the presence of suramin. In loose-patch clamp recordings, from intact neurohypophyses, suramin or PPADS produces an inhibition of action potential currents in a static bath, that can be mimicked by a hyperpolarization of the resting membrane potential (RMP). Correspondingly, in a static versus perfused bath there is a depolarization of the RMP of NHT, which was reduced by either suramin or PPADS. We measured an accumulation of ATP (3.7 +/- 0.7 microM) released from NHT in a static bath. Applications of either suramin or PPADS to a static bath decreased burst-stimulated capacitance increases in NHT. Finally, only vasopressin release from electrically stimulated intact neurohypophyses was reduced in the presence of Suramin or PPADS. These data suggest that there was sufficient accumulation of ATP released from the neurohypophysis during stimulations to depolarize its nerve terminals. This would occur via the opening of P2X2 and P2X3 receptors, inducing an influx of Ca2+. The subsequent elevation in [Ca2+](i) would further increase the stimulated release of only vasopressin from NHT terminals. Such purinergic feedback mechanisms could be physiologically important at most CNS synapses.

Purinergic P2X7 Receptor: A Pivotal Role in Surfactant Regulation?

P2X7 receptors (P2X7R) are specifically expressed in lung alveolar type I cells (AT1). We hypothesize that the activation of P2X7R on AT1 cells releases ATP and stimulates surfactant secretion from alveolar type II cells (AT2) via a paracrine manner. To test this hypothesis, we isolated primary rat alveolar epithelial cells by releasing them from lung tissues using enzymatic digestion. Immunophenotyping and cytometric analysis with cell specific markers showed 32% and 64% AT1 and AT2, respectively. Surfactant lipid secretion was measured on overnight cultured AT1 and AT2 cells in presence of 3′‐O‐(4‐benzoyl) benzoyl adenosine 5′‐triphosphate (BzATP), a known potent P2X7R agonist. BzATP significantly increased surfactant lipid secretion in the mixed cells in a dose dependent manner when compared to AT2 cells cultured alone and was significantly inhibited by P2X7R antagonist, Brilliant Blue G (BBG). However, when ATP was scavenged with apyrase and ADA, BzATP evoked surfactant secretion was significantly decreased. BzATP‐mediated secretion was also reduced in the presence of suramin and staurosporine, indicating the possible involvement of P2Y2R and downstream PKC‐mediated signaling pathway. Our results suggest that AT1 communicate in a paracrine fashion with AT2 cells in modulating surfactant secretion via P2X7R activation and ATP release.[NIH; HL 052146]

ZP120 causes relaxation by pre‐junctional inhibition of noradrenergic neurotransmission in rat mesenteric resistance arteries

ZP120 (Ac-RYYRWKKKKKKK-NH(2)), is a new partial nociceptin/orphanin FQ (NOP) receptor agonist with sodium-potassium sparing aquaretic effects. The mechanisms of vasodilatation of ZP120 were examined in rat mesenteric resistance arteries. Arterial segments (internal diameters 206+/-4 microm, n=224) were mounted in microvascular myographs for isometric tension recordings and electrical field stimulation (EFS). ZP120 and the endogenous NOP receptor ligand, N/OFQ, did not relax arteries contracted with noradrenaline or adenosine-triphosphate. EFS-evoked contractions were inhibited by a purinoceptor antagonist, suramin, and the alpha(1)-adrenoceptor antagonist prazosin. N/OFQ inhibited, concentration-dependently, EFS-evoked contractions with a maximal effect of 52+/-3% (n=8) at 1 microM. The maximal effect of 1 microM ZP120 was lower (27+/-5%, P<0.05, n=9) than for N/OFQ. Endothelial removal or pretreatment with capsaicin did not influence the vasodilator effects of ZP120 and N/OFQ. ZP120 and N/OFQ responses were preserved in the presence of suramin. The alpha(2)-adrenoceptor antagonist, rauwolscine, antagonized the effect of clonidine and brimonidine, but ZP120 and N/OFQ inhibition of EFS-evoked contraction was unaltered. The competitive NOP receptor antagonist, UFP-101 (10 microM), prevented the inhibitory effect of N/OFQ, but not ZP120 suggesting that N/OFQ and ZP120 have distinct modes of interaction with the NOP receptor. Our findings suggest that the vasodilator effect of ZP120 and N/OFQ in rat mesenteric resistance arteries is mediated by prejunctional inhibition of adrenergic neurotransmission. These properties, that promote diuresis and attenuate the cardiovascular consequences of increased sympathetic nerve activity, make ZP120 a promising drug candidate.