2,5-Di-t-butyl-1,4-benzohydroquinone induces endothelium-dependent relaxation of rat thoracic aorta

The aim of this work was to clarify the mechanism by which 2,5-di-t-butyl-1,4-benzohydroquinone (BHQ) induces relaxation of rat thoracic aorta. In particular, the role of endothelium-derived nitric oxide (NO) was investigated. BHQ concentration dependently (0.1-10 microM) relaxed rat aorta rings precontracted with phenylephrine. This effect was dependent on the intactness of the endothelium, suppressed by preincubation with 100 microM N(omega)-nitro-L-arginine methyl ester and antagonised by 3-30 microM methylene blue. The 10 microM BHQ-induced relaxation, however, was followed by the gradual and slow return to phenylephrine-induced tone. Superoxide dismutase (250 U/ml) increased the BHQ-induced relaxation, while preincubation with 3 mM diethyldithiocarbamate inhibited it in a time-dependent fashion. BHQ gave rise to superoxide anion formation which was markedly inhibited by the addition of superoxide dismutase (250 U/ml), either in the presence or in the absence of aorta rings. The non-specific blocker of Ca2+ channels, Ni2+, concentration dependently attenuated the BHQ relaxing effect. BHQ did not modify the relaxation induced by the NO donor 3-morpholino-sydnonimine in endothelium-deprived rings. In conclusion, BHQ induces endothelium-dependent relaxation and gives rise, by auto-oxidation, to the formation of superoxide anion. The former effect results from the enhanced synthesis of NO rather than from its enhanced biological activity; NO synthase is presumed to be stimulated by BHQ-induced activation of Ca2+ influx through Ni2+-sensitive Ca2+ channels.     

Endothelial nitric oxide-dependent vasorelaxant effect of isotirumalin, a dihydroflavonol from Derris urucu, on the rat aorta

The present work aimed to investigate the vasorelaxant effect of isotirumalin, a dihydroflavonol isolated from Derris urucu (Leguminosae). The vasorelaxant effect of isotirumalin was investigated in the rat aorta, in the presence and in the absence of a functional endothelium. The production of nitric oxide (NO) induced by isotirumalin was measured simultaneously with its vasorelaxation using carbon microsensors. In endothelium-intact aortic rings, isotirumalin induced a concentration-dependent vasodilator effect the concentration required to produce 30% of relaxation (pIC??=4.84±0.24) that was abolished in endothelium-denuded aortic rings or in the presence of Nω-nitro-L-arginine-methyl-ester (L-NAME; 300 μM). In addition, isotirumalin (100 μM) induced a simultaneous and significant increase on NO production, which was blunted in the presence of L-NAME. The present results demonstrate that isotirumalin is a vasodilator in the rat aorta and act by a mechanism dependent on the presence of a functional endothelium and on NO production.     

Dioclein, a new nitric oxide- and endothelium-dependent vasodilator flavonoid

In the present work, the vasorelaxant effect of dioclein, a new flavonoid isolated from Dioclea grandiflora (Leguminoseae), was investigated in the rat aorta. Dioclein induced a concentration-dependent relaxation in vessels pre-contracted with phenylephrine (IC(50)=1.3+/-0.3 microM), a response which was abolished after endothelium removal. Neither indomethacin (10 microM), an inhibitor of cyclo-oxygenase, nor atropine (1 microM), an antagonist of muscarinic receptors, modified the effect of dioclein. Dioclein (30 microM) induced a significant increase in guanosine 3':5'-cyclic monophosphate (cyclic GMP) levels in aortic rings with endothelium. The nitric oxide (NO) synthase inhibitor, N(G)-nitro-L-arginine-methyl-ester (L-NAME, 300 microM), strongly inhibited or abolished the relaxing effect and rise in cyclic GMP levels induced by dioclein. Furthermore, dioclein (30 microM) had no effect on the endothelium-independent relaxation produced by the NO donor, 3-morpholino-sydnonimine (SIN-1), while superoxide dismutase (100 U ml(-1)) significantly potentiated it. These results indicate that, in the rat aorta, dioclein induces a NO- and endothelium-dependent vasorelaxant effect, which is associated with cyclic GMP elevation. This vasorelaxation likely results from enhanced synthesis of NO rather than enhanced biological activity of NO.     

Gonadectomy eliminates endothelium-dependent diethylstilbestrol-induced relaxant effect in rat aorta

The effects of gender and castration of rats on diethylstilbestrol-induced, endothelium-dependent and endothelium-independent relaxation in rat aorta strips were studied. For this, male and female control and castrated rats were used. Diethylstilbestrol elicited a concentration-dependent (1-30 micromol/l) relaxation of isolated rat aorta. The effect was significantly higher in the presence of endothelium in aorta strips of the control group and also in female as compared with male rats. This effect is NO-dependent, since it is inhibited by N(G)-methyl-L-arginine. Castration of the rats suppressed the endothelium-dependent relaxation, and it was similar to that induced in the absence of endothelium. Acetylcholine-induced relaxation was not suppressed by castration. The acetylcholine-induced relaxation was decreased in aorta strips previously relaxed by diethylstilbestrol. There are no gender differences in the diethylstilbestrol-induced, endothelium-independent component of the relaxation, nor is it modified by the hormonal environment. Therefore, diethylstilbestrol-induced, endothelium-dependent relaxation in rat aorta strips is modulated by the hormonal status of the rats.     

Role of endothelium in the contractions induced by norepinephrine and clonidine in rat aorta

The inhibitory effects of endothelium-derived relaxing factor (EDRF) on the contractions induced by norepinephrine and clonidine in rat aorta were examined. Carbachol induced a relaxation of norepinephrine-induced contraction in rat aorta with endothelium. Removal of endothelium inhibited the carbachol-induced relaxation and increased the magnitude of norepinephrine-induced contraction. Quinacrine, a phospholipase A2 inhibitor, methylene blue, a guanylate cyclase inhibitor and tetraethylammonium, a potassium permeability inhibitor, inhibited carbachol-induced relaxation and augmented the magnitude of norepinephrine-induced contraction only when endothelium was present. Clonidine induced a contraction when endothelium was removed or muscle was treated with methylene blue. The contractions induced by norepinephrine and clonidine were equally sensitive to prazosin and equally less sensitive to yohimbine. Clonidine inhibited the norepinephrine-induced contraction, whereas it potentiated the angiotensin 11- or 12 mM K-induced contractions in the aorta with endothelium. The inhibitory effect of clonidine on the norepinephrine-induced contraction was reduced by endothelium-removal and by methylene blue but not by yohimbine. These results suggest that norepinephrine has a strong direct stimulating action and clonidine has a weak one on vascular smooth muscle cells possibly mediated by alpha 1-adrenoceptors, and their contractile effects are inhibited by the spontaneously released EDRF.     

Role of the endothelium in the vasodilator response of rat thoracic aorta to histamine

Despite their potent vasodilating action in vivo, acetylcholine and histamine often show a vasoconstricting action in vitro. As the endothelium has an important role in the vasodilating effect of acetylcholine, we investigated the possible role of the endothelium in the vasodilating effect of histamine in comparison to acetylcholine. Experiments were done on ring segments of rat thoracic aorta mounted for isometric tension measurements. We demonstrated that relaxation by histamine and acetylcholamine of pre-contracted rat aorta segments required the presence of endothelial cells. Acetylcholine acting on muscarinic receptors, and histamine acting on H₁-receptors seemed to initiate the production of mediator(s) from the endothelial cells, which leads to relaxation of the vascular smooth muscle cells. This production appeared to be depressed by ETYA and hydroquinone, and under hypoxic conditions.     

Trimebutine maleate relaxes the isolated rat thoracic aorta: The role of nitric oxide and L‐type calcium channels

Trimebutine maleate (TMB), a widely prescribed drug for functional gastrointestinal disorders, has been reported to regulate smooth muscle contractility by modulating multiple ion channel activities in the gastrointestinal tract. However, its action on isolated aorta has not yet been reported. The aim of the present study was to evaluate in vitro vasorelaxant properties and the underlying pharmacological mechanisms of TMB in isolated rat thoracic aortic rings. Vascular activity experiments were performed on thoracic aorta isolated from Sprague‐Dawley rats in vitro, including endothelium‐intact and endothelium‐denuded aortic rings. TMB (10^?10‐10^?5 mol/L) induced relaxation in endothelium‐intact aortic rings precontracted by phenylephrine with a potency similar to that of carbachol. TMB‐induced relaxation was not altered by glibenclamide and atropine in endothelium‐intact aortic rings. However, L‐NAME and endothelium denudation significantly reduced but not completely reversed the vasorelaxant effect of TMB. Also, TMB‐induced relaxation wasn't affected by diclofenac in endothelium‐intact aortic rings. TMB at 10^?5 mol/L significantly reduced the CaCl₂‐induced contractions in endothelium‐intact aortic rings stimulated with KCl, but not stimulated with phenylephrine under Ca²⁺free conditions. Moreover, TMB at 10^?5 mol/L effectively attenuated Bay‐K8644‐induced contractions in aortic rings. These results suggest that TMB‐induced relaxation was mediated by both endothelium‐dependent and endothelium‐independent manner in isolated rat thoracic aorta. The mechanism of TMB‐induced relaxation at low concentrations is partially related to NO‐ and endothelium‐dependent but unrelated to prostanoids formation. However, inhibition of Ca²⁺ influx through voltage‐operated calcium channels and L‐type Ca²⁺channel blocking effect appears to be involved in the mechanism of vasorelaxant effect of TMB at high concentrations.     

Inhibitory effect of the norlignan 2-(2'-hydroxy-4',6'-dimethoxyphenyl)-5-[(E)-propenyl]benzofuran from Krameria tomentosa on acetylcholine-induced relaxation of the rat aorta

In this work, we studied the effect of the norlignan 2-(2'-hydroxy-4',6'-dimethoxyphenyl)-5-[( E)-propenyl]benzofuran (DMPP) in the rat aorta. In aortic rings with intact endothelium, DMPP inhibited in a concentration-dependent manner the vasodilator effect produced by acetylcholine with an IC50 value of 31.2+/-6.3 microM. DMPP also inhibited basal nitric oxide production. In endothelium-denuded vessels DMPP was without effect whereas superoxide dismutase (SOD) was effective in potentiating responses to the NO donor SIN-1. Contractile effects of carbachol in guinea-pig ileum and trachea were unaffected by DMPP. It is concluded that DMPP inhibits the endothelium-dependent relaxation induced by acetylcholine in the rat aorta without affecting receptor or smooth muscle cells function. Decreased nitric oxide production by endothelial cells seems to be the mechanism involved in the inhibitory effect of DMPP.     

Endothelium-dependent vasorelaxation induced by black currant concentrate in rat thoracic aorta

We investigated the effect of black currant (BC) concentrate on smooth muscle in rat thoracic aorta. BC concentrate dose-dependently relaxed the norepinephrine (0.1 microM)-precontracted aorta, and the response was abolished after endothelium removal. Both oxyhemoglobin (1 microM), a nitric oxide (NO) scavenger, and IH-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one (ODQ, 0.5 microM), an inhibitor of guanylyl cyclase (GC), inhibited the relaxing effect of BC concentrate. NG-nitro-L-arginine methyl ester (L-NAME, 10 microM), a nitric oxide synthase (NOS) inhibitor, inhibited the relaxation, and the subsequent addition of L-arginine (1 mM), a NOS substrate, reversed the inhibitory effects of L-NAME. Neither indomethacin (10 microM), an inhibitor of cyclooxygenase, nor atropine (1 microM), an antagonist of muscarinic receptors, modified the effect of BC concentrate. Diphenhydramine (3 microM) and chlorpheniramine (2 microM), selective antagonists of H1-receptors, inhibited the relaxation, but cimetidine (0.3 mM), a selective antagonist of H2-receptors, did not affect the relaxation. These results indicate that, in the rat aorta, BC concentrate enhances synthesis of NO, which subsequently induces the endothelium-dependent vasorelaxation via the H1-receptors on the endothelium.     

Estradiol relaxes rat aorta via endothelium-dependent and -independent mechanisms

The effects of estrogen on arterial function are heterogeneous with respect to vessel and/or species. We have investigated 17beta-estradiol-induced relaxation in isolated rat aorta with regard to the role of the vascular endothelium and ionic mechanisms. Estrogen induced a concentration-dependent relaxation of 46.5 +/- 7.9% and 70.1 +/- 12.2% (10(-8) and 10(-7)M), which was reduced by endothelial denudation. Furthermore, L-nitroarginine methyl ester completely abrogated this effect; however, estradiol did not relax KCl-contracted rings. Tetraethyl ammonium (1 mmol/l) completely blocked estradiol-induced relaxation. Estradiol increased [cGMP] in isolated aortic rings via NO, but did not significantly affect NOS activity in endothelial cells. Thus, estrogen can relax rat aorta in vitro via both endothelium-dependent and -independent mechanisms involving the NO/cGMP and potassium channel signaling system.     

Nisoldipine increases the bioavailability of endothelial NO

Different observations suggest that dihydropyridine calcium antagonists alter endothelial NO release. Therefore, in a first step we investigated whether part of the nisoldipine (a dihydropyridine calcium antagonist with a possible selectivity for coronaries)-induced vasorelaxation was due to an NO release from the endothelium in porcine coronary arteries. Secondly, we directly measured whether nisoldipine increased NO release from rabbit aorta or the nisoldipine enantiomers (Bay R 1223, Bay R 1224) from rat aorta. Thirdly, we determined whether nisoldipine exerted antioxidative properties in segments of porcine aorta with intact endothelium. Blocking endothelial NO synthase with N-nitro-L-arginine resulted in a significant shift of the relaxation curve to higher concentrations. Accordingly, nisoldipine induced a concentration-dependent release of NO (direct electrochemical detection) from native endothelium which already started at a therapeutical level (1 nmol/l nisoldipine/6.5 +/- 1.2 nmol/l NO). To evaluate whether this effect was due to an antioxidative protection of NO, we examined the influence of nisoldipine on a hyperglycemia (30 mmol/l, 20 min)-induced reactive oxygen species release of vascular endothelium from porcine coronary arteries. Nisoldipine concentration-dependently reduced the reactive oxygen species release (>50%; 10 micromol/l). Moreover, a carbachol-induced NO release (rabbit aorta) which was significantly diminished by hyperglycemia was completely restored in the presence of nisoldipine (3 micromol/l). We conclude that nisoldipine increases the NO bioavailability which may result in an ameliorated endothelial function.     

Analysis of adenosine vascular effect in isolated rat aorta: possible role of Na+/K+-ATPase

The present experiments were undertaken in order to examine the effect of adenosine in isolated rat aorta, to investigate the possible role of intact endothelium and endothelial relaxing factors in this action and to determine which population of adenosine receptors is involved in rat aorta response to adenosine. Adenosine (0.1-300 microM) produced concentration-dependent (intact rings: pD2=4.39+/-0.09) and endothelium-independent (denuded rings: pD2=4.52+/-0.12) relaxation of isolated rat aorta. In the presence of high concentration of K+ (100 mM) adenosine-evoked relaxation was significantly reduced (maximal relaxation in denuded rings: control - 92.1+/-9.8 versus K+- 54.4+/-5.0). Similar results were obtained after incubation of ouabain (100 microM) or glibenclamide (1 microM). In K+-free solution, K+ (1-10 mM)-induced rat aorta relaxant response was significantly inhibited by ouabain (100 microM). Application of indomethacin (10 microM), NG-nitro-L-arginine (10 microM) or tetraethylammonium (500 microM) did not alter the adenosine-elicited effect in rat aorta. 8-(3-Chlorostyril)-caffeine (0.3-3 microM), a selective A2A-receptor antagonist, significantly reduced adenosine-induced relaxation of rat aorta in a concentration-dependent manner (pKB=6.57). Conversely, 1,3-dipropyl-8-cyclopentylxanthine (10 nM), an A1-receptor antagonist, did not affect adenosine-evoked dilatation. These results indicate that in isolated rat aorta, adenosine produces endothelium-independent relaxation, which is most probably dependent upon activation of smooth muscle Na+/K+-ATPase, and opening of ATP-sensitive K+ channels, to a smaller extent. According to receptor analysis, vasorelaxant action of adenosine in rat aorta is partly induced by activation of smooth muscle adenosine A2A receptors.     

Endothelin-3-induced relaxation of rat thoracic aorta: a role for nitric oxide formation.

1. Endothelin-3 (ET-3) at concentrations below those which caused contraction (30 nM) elicited endothelium-dependent relaxation followed by rebound contraction in rat isolated thoracic aorta. 2. Endothelin-1 also relaxed the rat aorta with a similar potency. 3. The nitric oxide (NO) synthase inhibitor, NG-nitro L-arginine, the radical scavenger, haemoglobin and the soluble guanylate cyclase inhibitor, methylene blue, each inhibited the ET-3-induced relaxation. 4. The calmodulin inhibitor, calmidazolium, considerably attenuated the relaxation caused by ET-3 without affecting that to nitroprusside. 5. Concentrations of ET-3 that were necessary to induce the relaxation also caused concentration-dependent elevation of guanosine 3':5'-cyclic monophosphate (cyclic GMP) levels. 6. NG-nitro L-arginine, haemoglobin, methylene blue, calmidazolium and removal of the endothelium completely abolished ET-3-stimulated cyclic GMP production. 7. These results suggest that ET-3 triggers NO formation possibly via ETB receptors on the endothelium to activate soluble guanylate cyclase, which in turn stimulates cyclic GMP production and smooth muscle relaxation. The enzyme contributing to the NO formation may be of the calcium/calmodulin-dependent, constitutive type.     

The release of a non-prostanoid inhibitory factor from rabbit bronchus detected by co-axial bioassay.

1. Methacholine relaxed phenylephrine-contracted aorta of the rat with the endothelium intact. This effect was inhibited by haemoglobin, methylene blue, gossypol, phenidone and L-NG-nitroarginine methyl ester (L-NAME). Rat aorta denuded of endothelium failed to relax in response to methacholine, histamine and the peptidoleukotrienes C4, D4 and E4. 2. Methacholine and histamine but not leukotrienes C4, D4 and E4 relaxed phenylephrine-contracted rat aorta without endothelium when surrounded by rabbit epithelium-intact bronchus. The muscarinic antagonist atropine antagonized the methacholine-induced relaxation. 3. Removal of the epithelium either mechanically or chemically, abolished methacholine-induced relaxation of rat aorta in the co-axial bioassay. These data indicate that the epithelium is responsible for the observed relaxant effect to methacholine and histamine. 4. The cyclo-oxygenase inhibitor, indomethacin, the phospholipase A2 inhibitor, mepacrine and the lipoxygenase inhibitor, nordihydroguaiaretic acid (NDGA), failed to inhibit methacholine-induced relaxation of rat aorta in the co-axial bioassay. This indicates that the epithelium-derived inhibitory factor (EpDIF) is not a product of the cyclo-oxygenase or lipoxygenase pathway or a product derived from activation of phospholipase A2. 5. Haemoglobin, methylene blue, phenidone, gossypol and L-NAME failed to inhibit the relaxation of rat aorta in the co-axial bioassay. These results demonstrate that EpDIF detected in the co-axial bioassay is not endothelium-derived relaxing factor (EDRF) or nitric oxide. Similarly, catalase was without effect. 6. EpDIF is unlikely to be a peptide since papain and alpha-chymotrypsin failed to alter the methacholine-induced relaxation of rat aorta in the co-axial bioassay.(ABSTRACT TRUNCATED AT 250 WORDS)     

Analysis of Adenosine Vascular Effect in Isolated Rat Aorta: Possible Role of Na+/K+‐ATPase

Abstract: The present experiments were undertaken in order to examine the effect of adenosine in isolated rat aorta, to investigate the possible role of intact endothelium and endothelial relaxing factors in this action and to determine which population of adenosine receptors is involved in rat aorta response to adenosine. Adenosine (0.1–300 μM) produced concentration‐dependent (intact rings: pD₂=4.39±0.09) and endothelium‐independent (denuded rings: pD₂=4.52±0.12) relaxation of isolated rat aorta. In the presence of high concentration of K⁺ (100 mM) adenosine‐evoked relaxation was significantly reduced (maximal relaxation in denuded rings: control – 92.1±9.8 versus K⁺– 54.4±5.0). Similar results were obtained after incubation of ouabain (100 μM) or glibenclamide (1 μM). In K⁺‐free solution, K⁺ (1–10 mM)‐induced rat aorta relaxant response was significantly inhibited by ouabain (100 μM). Application of indomethacin (10 μM), N^G‐nitro‐L‐arginine (10 μM) or tetraethylammonium (500 μM) did not alter the adenosine‐elicited effect in rat aorta. 8‐(3‐Chlorostyril)‐caffeine (0.3–3 μM), a selective A_2A‐receptor antagonist, significantly reduced adenosine‐induced relaxation of rat aorta in a concentration‐dependent manner (pK_B=6.57). Conversely, 1,3‐dipropyl‐8‐cyclopentylxanthine (10 nM), an A₁‐receptor antagonist, did not affect adenosine‐evoked dilatation. These results indicate that in isolated rat aorta, adenosine produces endothelium‐independent relaxation, which is most probably dependent upon activation of smooth muscle Na⁺/K⁺‐ATPase, and opening of ATP‐sensitive K⁺ channels, to a smaller extent. According to receptor analysis, vasorelaxant action of adenosine in rat aorta is partly induced by activation of smooth muscle adenosine A_2A receptors.     

Endogenous nitric oxide attenuates beta-adrenoceptor-mediated relaxation in rat aorta

1. Divergent evidence suggests that the intracellular signalling pathways for beta-adrenoceptor-mediated vascular relaxation involves either cAMP/protein kinase (PK) A or endothelial nitric oxide (NO) release and subsequent activation of cGMP/PKG. The present study identifies the relative roles of NO and cAMP, as well as dependence on the endothelium for beta-adrenoceptor-mediated relaxation of rat isolated aortas. 2. Cumulative concentration-response curves to isoprenaline (0.01-3 micromol/L) in phenylephrine (0.1 micromol/L)-preconstricted endothelium-intact and -denuded aortas were constructed. Isoprenaline-mediated relaxation was partially reduced by endothelium removal and the presence of the NO synthase inhibitor N(G)-monomethyl-L-arginine (0.1 mmol/L), but not by the cAMP antagonist (Rp)-cyclic adenosine-3',5'-monophosphorothioate (Rp-cAMPS; 0.5 mmol/L). 3. In contrast, in endothelium-denuded aortas, the isoprenaline-mediated relaxation was inhibited by Rp-cAMPS and this inhibition was lost in the presence of the NO donor sodium nitroprusside (1 nmol/L). This effect was not due to phosphodiesterase (PDE) activity because the non-selective PDE inhibitor 3-isobutyl-1-methylxanthine (1 micromol/L) failed to affect the isoprenaline vasorelaxant response. 4. The K(+) channel blocker tetraethylammonium (TEA; 1 mmol/L) attenuated isoprenaline-induced relaxation in endothelium-denuded aorta, but its effect was non-additive with Rp-cAMPS, suggesting that the K(+) channel component may involve cAMP. In endothelium-intact aortas, TEA but not Rp-cAMPS reduced isoprenaline relaxation, suggesting an additional non-cAMP component. 5. These findings suggest that beta-adrenoceptors induce vascular smooth muscle relaxation by acting through the NO-cGMP pathway and, when that is disrupted by endothelium removal or the presence of an NO synthase inhibitor, the cAMP pathway in smooth muscles is used. The lack of cAMP participation in endothelium-intact vessels may be because NO suppresses or overrides the cAMP effect.     

A novel in vitro endothelium-dependent vascular relaxant effect of Apocynum venetum leaf extract

1. In the present study, a novel in vitro vascular relaxant effect of Apocynum venetum leaf extract (AVLE; also called 'Luobuma'), obtained from a traditional Chinese medicinal herb with known antihypertensive effects, is reported in isometric contraction studies of rat aorta and superior mesenteric artery. At low concentrations (0.3-10 microg/mL), AVLE had no effect on the resting tension of either blood vessel and caused relaxation in agonist-precontracted vessels with functionally intact endothelium. 2. We demonstrated pharmacologically that the AVLE-induced vasorelaxation was mediated selectively by the endothelial cells in both blood vessels. Using NG-nitro-L-arginine methyl ester (L-NAME) and a low concentration of KCl (15 mmol/L), we also demonstrated that AVLE acted by releasing endothelium-derived relaxation factors; nitric oxide (NO) in the rat aorta and NO plus endothelium-derived hyperpolarizing factor in the rat mesenteric artery. 3. The vascular relaxation following brief exposure to AVLE appeared to persist even after subsequent prolonged washout; this was manifested as an attenuated contraction to subsequent application of phenylephrine (PE) compared with the PE-induced contraction after exposure to carbachol (CCh) and subsequent similar washout. The addition of L-NAME at this point in the absence of AVLE totally restored the contraction to PE, suggesting that enzymatic generation of endothelial NO persisted even after brief exposure to AVLE. 4. Unlike the endothelium-dependent NO-mediated relaxation induced by CCh, which is mediated by endothelial muscarinic receptors (and inhibited by atropine), the relaxation induced by AVLE was not inhibited by atropine and, thus, was not mediated by muscarinic receptors. However, similar to CCh-induced relaxation, AVLE-induced relaxation was associated with the activation of K+ channels. 5. These results provide a strong scientific basis for the folk use of AVLE decoction for antihypertensive therapy in traditional Chinese medicine.     

Effects of hydrazine derivatives on vascular smooth muscle contractility,blood pressure and cGMP production in rats: comparison with hydralazine

Hydralazine is a hydrazine derivative used clinically as a vasodilator and antihypertensive agent. Despite numerous studies with the drug, its mechanism of action has remained unknown; guanylate cyclase activation and release of endothelial relaxing factors are thought to be involved in its vasodilator effect. Other hydrazine derivatives are known to stimulate guanylate cyclase and could therefore share the vasodilator activity of hydralazine, although such possibility has not been assessed systematically. In the present study, hydralazine, hydrazine, phenylhydrazine, and isoniazid were evaluated for vascular smooth muscle relaxation in rat aortic rings with and without endothelium, as well as after incubation with the guanylate cyclase inhibitor methylene blue. They were also tested for enhancement of cyclic guanosine monophosphate (cGMP) production by cultured rat aortic smooth muscle cells and for hypotension in the anesthetized rat. All hydrazines relaxed aortic rings, an action unaffected by endothelium removal and, in all cases except hydralazine, antagonized by methylene blue. Only phenylhydrazine increased cGMP production and only hydralazine markedly lowered blood pressure. It was concluded that hydralazine vascular relaxation is independent of endothelium and is not related to guanylate cyclase activation. The other hydrazines studied also elicit endothelium-independent relaxation, but the effect is related to guanylate cyclase. The marked hypotensive effect of hydralazine contrasts with its modest relaxant activity and is not shared by the other hydrazines. The fact that hydrazine and isoniazid produce methylene blue-sensitive relaxation, yet do not enhance cGMP production suggests the need for activating factors present in aortic rings but not in isolated cells.     

L-arginine induces relaxation of rat aorta possibly through non-endothelial nitric oxide formation.

1. The relaxation of rings of rat thoracic aorta induced by L-arginine and its derivatives was investigated. 2. L-Arginine (0.3-100 microM), but not D-arginine, induced relaxation of the arteries, which was detectable after 2 h and maximal after 4-6 h on its repeated application; it was endothelium-independent. 3. L-Arginine methyl ester, N alpha-benzoyl L-arginine and L-homo-arginine had essentially similar effects to those of L-arginine. 4. NG-nitro L-arginine methyl ester (L-NAME, 3 microM), NG-nitro L-arginine (L-NNA, 1 microM) and NG-monomethyl L-arginine (L-NMMA, 10 microM), inhibitors of nitric oxide (NO) formation from L-arginine, inhibited or reversed the L-arginine-induced relaxation, irrespective of the presence or absence of the endothelium. In contrast, NG-nitro D-arginine was without effect. 5. Haemoglobin (Hb, 10 nM) and methylene blue (MB, 0.3 microM) inhibited or reversed the L-arginine-induced relaxation. 6. L-Arginine (1-100 microM), but not D-arginine, increased guanosine 3':5'-cyclic monophosphate (cyclic GMP) levels in the tissues that relaxed in response to L-arginine. This effect of L-arginine was suppressed by Hb (3 microM), MB (1 microM) and L-NAME (100 microM). Removal of the endothelium did not significantly alter the L-arginine-induced cyclic GMP production. 7. These results suggest that L-arginine itself caused a slowly developing relaxation of rat aorta, possibly via formation of NO by an endothelium-independent mechanism.     

Potassium channels-mediated vasorelaxation of rat aorta induced by resveratrol

Resveratrol, a phenolic substance present in grapes and a variety of medical plants, has been reported to induce vasorelaxation, however the mechanisms are uncertain. In this paper we investigate the possible participation of K(+) channels in the endothelium-independent vasodilatation of rat aorta induced by resveratrol. Resveratrol induced concentration-dependent relaxation of rings with endothelium and without endothelium. We used different potassium channel inhibitors to determine whether the K(+) channels mediated endothelium-independent relaxation of rat aorta induced by resveratrol. Highly selective blocker of ATP-sensitive K(+) channels, glibenclamide, as well as non-selective blockers of K(+) channels, tetraethylammonium, did not block resveratrol-induced relaxation of rat aortic rings. Charybdotoxin, a blocker of calcium-sensitive K(+) channels did not affect the resveratrol-induced relaxation. 4-Aminopiridine, non-selective blocker of voltage-gated K(+) (Kv) channels, and margatoxin that inhibits Kv1 channels abolished relaxation of rat aortic rings induced by resveratrol. In conclusion, we have shown that resveratrol potently relaxed rat aortic rings with denuded endothelium. It seems that 4-aminopiridine and margatoxin-sensitive K(+) channels located in the smooth muscle of rat aorta mediated this relaxation.