Biomedical subjects

Lusiane M Bendhack

Publications and source records attributed to Lusiane M Bendhack.

16 recordsLinked to original sources

PubMed2006

Involvement of non-selective Ca2+ channels in the contraction induced by alkalinization of rat anococcygeus muscle cells.

Intracellular pH is a modulator of cellular functions such as smooth muscle contraction. Changes in cytosolic Ca(2+) concentration ([Ca(2+)](c)) associated with contraction are brought about by Ca(2+) influx and release from the sarcoplasmic reticulum, and alterations in the intracellular pH can affect both processes. In this work, therefore, we have investigated the Ca(2+) influx pathway that contributes to the contraction induced by the alkalinizing agent NH(4)Cl in the rat anococcygeus smooth muscle. For this purpose, we measured the isometric tension in muscle preparations, and [Ca(2+)](c) was measured on isolated cells loaded with 5 micromol/l FURA2/AM by using the ratio 340/380 nm. NH(4)Cl (10 mmol/l) induced a larger increase in [Ca(2+)](c) (100%) when compared with the [Ca(2+)](c) increase induced by 0.1 micromol/l phenylephrine (57.0+/-12.3% n=4). Incubation of the muscle preparations for 1 min in Ca(2+)-free medium reduced the contractions induced by 10 mmol/l NH(4)Cl to 11.5+/-5.1% (n=5), when compared with the contractions induced in 2.5 mmol/l Ca(2+) solution (100%). After 3 min in Ca(2+) free medium, contractions stimulated with NH(4)Cl were almost abolished (0.6+/-0.4%, n=5). In the same way, incubation with 10 micromol/l 1-[beta-[3[(4-methoxyphenyl)propoxyl]-4-methoxy-phenetyl]-1H-imidazole hydrochloride (SKF96365), a non-selective Ca(2+) channels, reduced the contractions stimulated with NH(4)Cl to 47.6+/-6.7% (n=7). On the other hand, 1 micromol/l verapamil, a voltage-operated Ca(2+) channel blocker and 0.05 micromol/l calphostin C, a protein kinase-C inhibitor, did not alter the contractions induced by NH(4)Cl. On isolated cells, [Ca(2+)](c) was reduced to 72.2+/-1.7% (n=4) by 10 micromol/l SKF96365. Taken together, our results suggest that NH(4)Cl induces contraction of rat anococcygeus smooth muscle cells, as well as [Ca(2+)](c) increase due to Ca(2+) influx through non-selective Ca(2+) channels.

Ammonium Chloride↗

PubMed2006

Comparison of the mechanisms underlying the relaxation induced by two nitric oxide donors: sodium nitroprusside and a new ruthenium complex.

We studied the mechanisms involved in the relaxation induced by nitric oxide (NO) donors, ruthenium complex ([Ru(terpy)(bdq)NO(+)](3+)-TERPY) and sodium nitroprusside (SNP) in denuded rat aorta. Both NO donors induced vascular relaxation independent of the agonist used in the pre-contraction. [Ru(terpy)(bdq)NO(+)](3+) and SNP activated guanylyl cyclase (GC) and K(+) channels. The production of cGMP induced by [Ru(terpy)(bdq)NO(+)](3+) - was higher than that obtained with SNP. The combination of GC inhibitor with K(+)channels blocker almost abolished the relaxation induced by the NO donors. The extracellular NO scavenger oxyhemoglobin reduced the potency without changing the maximum effect (Emax) of both NO donors. By using specific NO species scavengers, hydroxocobalamin and l-cysteine, we have identified the contribution of free radical NO (NO()) and nytroxil anion (NO(-)), respectively, to the rat aorta relaxation induced by both NO donors. The selective scavengers for NO() and NO(-) reduced the potency but not the Emax of [Ru(terpy)(bdq)NO(+)](3+). However, the NO(-) scavenger had no effect on the relaxation induced by SNP and NO() scavenger reduced only the potency to SNP. The inhibition of sarcoplasmic reticulum Ca(2+)-ATPase reduced only the potency of SNP without effect on the relaxation induced by [Ru(terpy)(bdq)NO(+)](3+). Our results demonstrate that both NO donors induce relaxation by activating the GC and K(+) channels. The NO() is the unique NO specie involved in the SNP-relaxation. On the other hand, the relaxant effect of [Ru(terpy)(bdq)NO(+)](3+) involves both NO() and NO(-), that produce higher concentration of cGMP. The inhibition of sarcoplasmic reticulum Ca(2+)-ATPase reduces the relaxation induced by SNP but it did not alter the relaxation induced by [Ru(terpy)(bdq)NO(+)](3+).

Animals↗

PubMed2006

Cross-talk between the sarcoplasmic reticulum and the mitochondrial calcium handling systems may play an important role in the regulation of contraction in anococcygeus smooth muscle.

Mitochondrial Ca(2+) and its relation with the contraction induced by phenylephrine was investigated. In normal Ca(2+), carbonyl cyanide p-(trifluoro-methoxy)phenyl-hydrazone (FCCP) and oligomycin produced contraction similar to that promoted by phenylephrine. Phenylephrine-induced contraction was reduced by FCCP+oligomycin. In Ca(2+)-free, FCCP+oligomycin did not induce contraction. Response to FCCP+oligomycin was reduced upon Ca(2+) repletion and this response was lower than that to phenylephrine. Ca(2+) concentration was increased by FCCP+oligomycin. Since a profuse net of sarcoplasmic reticulum encloses mitochondria, a cross-talk between the two organelles may play an important role in the phenylephrine-induced contraction in presence of Ca(2+) encountered in both sarcoplasmic reticulum and extracellular medium of anococcygeus cells.

Animals↗

PubMed2006

Characterization of the mechanisms of action and nitric oxide species involved in the relaxation induced by the ruthenium complex.

Nitric oxide (NO) plays an important role in the control of vascular tone. NO donors have therapeutic use and the most used NO donors, nitroglycerin and sodium nitroprusside have problems in their use. Thus, new NO donors have been synthesized to minimize these undesirable effects. Nytrosil ruthenium complexes have been studied as a new class of NO donors. trans-[RuCl([15]aneN(4))NO](2+), induces vasorelaxation only in presence of reducing agent. In this study, we characterized the mechanisms of vasorelaxation of trans-[RuCl([15]aneN(4))NO](2+) in denuded rat aorta and identified which NO forms are involved in this relaxation. We also evaluated the effect of this NO donor in decreasing the cytosolic Ca(2+) concentration ([Ca(2+)]c) of the vascular smooth muscle cells. Vasorelaxation to trans-[RuCl([15]aneN(4))NO](2+) (E(max): 101.8 +/- 2.3%, pEC(50): 5.03 +/- 0.15) was almost abolished in the presence of the NO* scavenger hydroxocobalamin (E(max): 4.0 +/- 0.4%; P < 0.001) and it was partially inhibited by the NO(-) scavenger L-cysteine (E(max): 79.9 +/- 6.9%, pEC(50): 4.41 +/- 0.06; P < 0.05). The guanylyl cyclase inhibitor ODQ reduced the E(max) (57.7 +/- 4.0%, P < 0.001) and pEC(50) (4.21 +/- 0.42, P < 0.01) and the combination of ODQ and TEA abolished the response to trans-[RuCl([15]aneN(4))NO](2+). The blockade of voltage-dependent (K(v)), ATP-sensitive (K(ATP)), and Ca(2+)-activated (K(Ca) K(+) channels reduced the vasorelaxation induced by trans-[RuCl([15]aneN(4))NO](2+). This compound significantly reduced [Ca(2+)]c (from 100% to 85.9 +/- 3.5%, n = 4). In conclusion, our data demonstrate that this NO donor induces vascular relaxation involving NO* and NO(-) species, that is associated to a decrease in [Ca(2+)]c. The mechanisms of vasorelaxation involve guanylyl cyclase activation, cGMP production and K(+) channels activation.

Animals↗

PubMed2005

Decreased contraction to phenylephrine by ouabain in 2K-1C rat aorta is modulated by the endothelium.

The effects of ouabain were studied on the contraction stimulated with phenylephrine or KCl in intact endothelium and denuded aortic rings isolated from normotensive (2K) and renal hypertensive 2 kidney-1clip (2K-1C) rats. Ouabain did not change the basal tone of aortic rings. Ouabain (1 nmol/l) had no effect on the contraction to phenylephrine in all the artery groups studied. Ouabain (10 nmol/l) decreased the E(max) to phenylephrine in intact endothelium arteries from 2K-1C. By contrast, ouabain (10 nmol/l) had no effect on the contraction to KCl. Ouabain induced membrane depolarization measured by confocal image with Di-4-ANEPPS dye, that was greater in 2K than in 2K-1C rat aorta smooth muscle cells. In conclusion, ouabain (10 nmol/l) decreased the contractile responses to phenylephrine only in 2K-1C rat aortic rings with intact endothelium. Interestingly, 10 nmol/l ouabain depolarizes the smooth muscle cells but this depolarization level is not enough to alter the phenylephrine or KCl-induced contractions. Our results indicate that the endothelium modulates the vascular action of ouabain.

Animals↗

PubMed2005

The complex trans-[RuCl([15]aneN4NO]2+ induces rat aorta relaxation by ultraviolet light irradiation.

The aim of the present study was to investigate the possible endogenous storage of photosensitive nitric oxide, and also to examine the relaxant effect of NO released from the compound by UV light irradiation. Aorta was isolated from rats and the endothelium was mechanically removed. Denuded aortic rings pre-contracted with prostaglandin F(2alpha) responded with relaxation to UV light irradiation. The first stimulation produced the greatest response that decreased until complete disappearance. After this, the addition of the compound in the absence of light did not produce any response. However, in the presence of UV light irradiation, the complex trans-[RuCl([15]aneN4)NO]2+ induced 100% relaxation. After incubation with the nitric oxide scavenger, oxyhaemoglobin, this relaxation was completely abolished. In PGF2(2alpha)-pre-contracted aortas, the time to reach maximum relaxation was only 50s. Taken together, these results suggest that preformed endogenous nitric oxide stores exist in the denuded rat aorta, and that they are sensitive to UV light. The photo-induction of the complex trans-[RuCl([15]aneN4NO]2+ induces complete aorta relaxation, which is due to release of nitric oxide in the extracellular medium.

Animals↗

PubMed2005

Effects of voltage-gated Na+ channel toxins from Tityus serrulatus venom on rat arterial blood pressure and plasma catecholamines.

Scorpion toxins interact with ionic channels of excitable cells, leading to a massive release of neurotransmitters. Voltage-gated Na+ channel toxins are mainly responsible for the toxic effects of scorpion envenoming and can be classified into two classes: alpha- and beta-neurotoxins. TsTX-V and TsTX-I from Tityus serrulatus venom (TsV) are, respectively, examples of these toxins. In this work, we compared the effects of these toxins on mean arterial pressure (MAP) and catecholamines release in rats. Toxins were isolated by ion exchange chromatography (TsTX-I) followed by RP-HPLC (TsTX-V). All experiments were performed on conscious unrestrained rats previously catheterised. The toxins (15 and 30 microg/kg) and TsV (50 and 100 microg/kg) were injected intravenously. MAP was continuously monitored through femoral catheter. Epinephrine (E) and norepinephrine (NE) levels were determined by RP-HPLC with electrochemical detection, at 10 min before and 2.5, 30 and 90 min after treatments. Maximal pressor effects were observed at 2.5-3.5 min. TsV induced intense long lasting increase in MAP, as did TsTX-I. TsTX-V showed the lowest pressor effects. TsV showed the highest effects on catecholamines release, followed by TsTX-I and TsTX-V with maximal effect at 2.5 min, followed by a gradual reduction, however remaining higher than controls. Although both toxins act on Na+ channels, TsTX-I displayed significant and more intense effects on catecholamines release and blood pressure than TsTX-V. It seems that the toxicity of TsTX-V is not related only with its ability to release catecholamines, indicating that other neurotransmitters, may be involved in its toxicity.

Animals↗

PubMed2005

Activation of alpha2-adrenoceptors is necessary to induce nitric oxide release in isoprenaline-induced relaxation.

The aim of this study was to investigate the role of the contractile agent on the relaxation induced by isoprenaline and the contribution of nitric oxide (NO) and cGMP to this relaxation. These studies were conducted in intact endothelium or denuded aortas contracted with the EC50 of norepinephrine (NE) or phenylephrine (Phe), and the relaxation induced by isoprenaline (non-selective beta-adrenoceptor agonist) or forskolin (activator of adenylyl-cyclase) was studied. The maximum effect (Emax) and pD2 were analysed. Isoprenaline and forskolin-induced relaxation were not changed by the endothelium removal in both NE and Phe-contracted aortas. However, L-NAME reduced the relaxation induced by isoprenaline (Emax from 94.48+/-2.30%, n=7 to 66.17+/-11.73%, n=7; pD2 from 7.56+/-0.10 to 6.08+/-0.15) only in NE-contracted aortas. The pD2 of isoprenaline was also reduced by ODQ (6.57+/-0.13), but not the Emax. The inhibitory effects of L-NAME and ODQ were reversed by yohimbine. L-NAME, ODQ and oxyhemoglobin had no effect on the relaxation induced by isoprenaline in Phe-contracted aortas. Taken together, these results suggest that norepinephrine, a non-selective alpha-adrenoceptor agonist can also activate alpha2-adrenoceptors sensitive to yohimbine in the endothelial cells, activating the NO-synthase and cGMP production which would potentiate the relaxation induced by isoprenaline. However, this pathway is not activated with Phe, the selective alpha1-adrenoceptors agonist.

Adrenergic beta-Agonists↗

PubMed2004

Atorvastatin enhances sildenafil-induced vasodilation through nitric oxide-mediated mechanisms.

Statins have cholesterol-independent effects including an increased vascular nitric oxide (NO) activity and are commonly used by patients with cardiovascular disease. Such patients frequently have erectile dysfunction, which may be treated with sildenafil, a selective inhibitor of phosphodiesterase type 5. Since statins and sildenafil can activate the NO-cGMP pathway, we investigated whether pre-treatment with atorvastatin (0, 5 and 30 mg/kg/day) for 2 weeks affects sildenafil (1 pM-100 mM)-induced relaxation of aortic rings isolated from Wistar rats. We also examined the hemodynamic consequences of this interaction in Wistar rats. Plasma nitrite/nitrate (NOx) concentrations were determined using an ozone-based chemiluminescence assay. While pre-treatment with atorvastatin increased the potency of sildenafil-induced vasorelaxation (P<0.01), no differences were observed in the maximum sildenafil-induced relaxation. Pre-incubation of aortic rings with NG-nitro-L-arginine methyl ester (L-NAME) reversed atorvastatin-induced increase in the potency of sildenafil relaxation. In addition, pre-treatment with atorvastatin enhanced plasma NOx concentrations and sildenafil-induced hypotension and tachycardia (all P<0.05). These results suggest that atorvastatin increases the vascular sensitivity to sildenafil through NO-mediated mechanisms.

Analysis of Variance↗

PubMed2004

Changes in the vascular beta-adrenoceptor-activated signalling pathway in 2Kidney-1Clip hypertensive rats.

1. beta-Adrenoceptor (beta-AR)-mediated vasodilation, which plays an important physiological role in the regulation of vascular tone, is decreased in two-kidney, one clip (2K-1C) renal hypertension. In this study, downstream pathways related to vascular beta-AR activation were evaluated in 2K-1C rats. 2. Relaxation responses to isoprenaline, forskolin and 8-Br-cAMP were diminished in aortas without endothelium from 2K-1C when compared to those in normotensive two kidney (2K). Basal adenosine-3',5'-monophosphate (cAMP), as well as isoprenaline-induced increase in cAMP levels, was not different between 2K and 2K-1C aortas. 3. Contractile responses to caffeine, after depletion and reloading of intracellular Ca(2+) stores, were greater in 2K-1C than in 2K. The presence of isoprenaline during the Ca(2+)-reloading period abolished the differences between groups by increasing caffeine contraction in 2K without changing this response in 2K-1C aortas. Inhibition of the sarcolemmal Ca(2+)ATPase with thapsigargin markedly attenuated isoprenaline vasodilation in both 2K and 2K-1C and abolished the differences between groups. 4. Blockade of ATP-sensitive K(+) channels (K(ATP)) channels with glibenclamide significantly decreased isoprenaline vasodilation in 2K-1C without affecting this response in 2K. Both vascular gene and protein expression of protein kinase A (PKA), as well as phosphoserine-containing proteins, were increased in 2K-1C vs 2K rats. 5. In conclusion, decreased isoprenaline vasodilation in 2K-1C hypertensive rats is related to impaired modulation of the sarcolemmal Ca(2+)ATPase activity. Moreover, K(ATP) channels may play a compensatory role on isoprenaline-induced relaxation in renal hypertension. Both Ca(2+)ATPase and K(ATP) channel functional alterations, associated with decreased beta-AR vasodilation, are paralleled by an upregulation of protein kinase A (PKA) and phosphoserine proteins expression.

8-Bromo Cyclic Adenosine Monophosphate↗

PubMed2004

Ca2+-activated K+ channels underlying the impaired acetylcholine-induced vasodilation in 2K-1C hypertensive rats.

We tested the hypothesis that an abnormal function of K(+) channels in vascular smooth muscle cells plays a key role in the impaired acetylcholine (ACh) vasodilation in aortas from two kidney-one clip (2K-1C) hypertensive rats and further investigated the K(+) channel subtype involved in this altered response. ACh-induced endothelium-dependent relaxation was assessed in aortic rings from 2K-1C and normotensive two kidney (2K) rats. Glibenclamide, an ATP-sensitive K(+) channel blocker, did not inhibit ACh-induced relaxation in aortic rings from 2K or 2K-1C rats. The voltage-dependent K(+) channels inhibitor 4-aminopyridine attenuated ACh-induced relaxation in both groups. Charybdotoxin and iberiotoxin, blockers of Ca(2+)-sensitive (K(Ca)) and large-conductance K(Ca) (BK(Ca)) channels, respectively, reduced ACh-induced relaxation in aortic rings from 2K rats without affecting this response in those from 2K-1C rats, abolishing the differences between groups. ACh-induced relaxation in vessels from both 2K and 2K-1C rats was unaffected by apamin, a small-conductance K(Ca) blocker. NS1619 [1,3-dihydro-1-[2-hydroxy-5-(trifluoromethyl)phenyl]-5-(trifluoromethyl)-2H-benzimidazol-2-one], an activator of K(Ca), induced a smaller vasodilation in endothelium-denuded aortic rings from 2K-1C rats compared with those from 2K rats. Iberiotoxin reduced sodium nitroprusside-induced relaxation in endothelium-denuded aortic rings from 2K without affecting this response in those from 2K-1C rats. The inhibition of Na(+),K(+)-ATPase with ouabain had no effects on ACh-induced relaxation in aortic rings from 2K-1C or 2K rats. These data indicate that a deficient functional activity of BK(Ca) channels plays a key role in the impaired ACh vasodilation in aortas from 2K-1C rats.

Acetylcholine↗

PubMed2004

A macrocyclic nitrosyl ruthenium complex is a NO donor that induces rat aorta relaxation.

The vasorelaxation induced by a nitrosyl macrocyclic ruthenium complex, proposed as a new nitric oxide (NO) carrier, was studied in rat isolated aorta. The compound trans-[RuCl([15]aneN4)NO]2+ was characterized by elemental analysis, UV-visible spectrum, and infrared spectrum. Based on the electrochemical process, the reduction of the compound was followed by NO release, which was also observed using norepinephrine as a reducing agent and NO released was analyzed by a sensor. Vasorelaxation induced by this NO donor was studied and compared to those obtained with sodium nitroprusside (SNP). The relaxation induced by the compound was concentration-dependent in denuded rat aortas and occurred only in pre-contracted arteries with norepinephrine. The macrocyclic compound induced relaxation with a similar efficacy as SNP, although the potency of SNP was slightly greater. The time to reach maximum relaxation (595 s) was longer than that of SNP (195 s). Relaxation was completely abolished by oxyhemoglobin, a known NO scavenger.

Animals↗

PubMed2004

Clonidine induces rat aorta relaxation by nitric oxide-dependent and -independent mechanisms.

The alpha1- and alpha2-adrenoceptors coexist in vascular smooth muscle cells producing vascular contraction and relaxation. This study was designed to investigate which is the mechanism activated by clonidine in the rat aorta, and the endothelial factors possibly involved in the relaxation induced by clonidine. The alpha2-adrenoceptors agonist clonidine relaxed rat aortas pre-contracted with phenylephrine, with or without endothelium. In non-contracted denuded arteries, clonidine produced contractions instead of relaxation. In intact endothelium aortic rings, clonidine induced greater relaxation than in denuded aortic rings. In aortas with intact endothelium, the NO-synthase inhibitor L-NAME (10 micromol/L) and the NO-scavenger hemoglobin (10 micromol/L) reduced the relaxation to clonidine. On the other hand, indomethacin (10 micromol/L) failed to alter the relaxation induced by clonidine. These results suggest the participation of NO, but not prostacyclin in clonidine-induced relaxation. In aortic rings pre-contracted with KCl (60 mmol/L) the relaxation induced by clonidine was abolished; however, the K+ channel blockers glibenclamide (K(ATP)), tetraethylamonium (K(Ca)), and the combination of apamin and charybdotoxin (K(Ca)) did not change the relaxation induced by clonidine. The relaxation induced by clonidine on PGF2alpha-contracted arteries was not affected by prazosin. However, in the absence of prazosin, clonidine had an additional contractile effect in PGF2alpha-contracted arteries. In conclusion, our results show that in rat aorta clonidine can activate alpha2-adrenoceptors in the smooth muscle cells and alpha2-adrenoceptors in the endothelial cells that activates NO production, but not prostacyclin and/or EDHF. In the absence of phenylephrine and prazosin, clonidine can also activate alpha1-adrenoceptors and rat aorta contraction.

Adrenergic alpha-Agonists↗

PubMed2004

Relaxation induced by acetylcholine involves endothelium-derived hyperpolarizing factor in 2-kidney 1-clip hypertensive rat carotid arteries.

Acetylcholine induced relaxation in a concentration-dependent way in isolated phenylephrine-contracted carotid artery rings from normotensive two-kidney (2K) and hypertensive two-kidney one-clip (2K-1C) rats. In the presence of the nitric oxide (NO) synthase inhibitor NG-nitro-L-arginine (L-NOARG, 100 micromol/l), the relaxation stimulated with acetylcholine was blocked in 2K arteries. However, in 2K-1C arteries, the relaxation was only partially inhibited. Indomethacin (3 micromol/l) had no effect in both groups. In 2K arteries, the combination of L-NOARG and indomethacin had similar effects to L-NOARG alone. On the other hand, in 2K-1C arteries, indomethacin further inhibited the maximum effect induced by acetylcholine. Endothelium-dependent relaxation induced by acetylcholine was markedly reduced in 2K arteries contracted with 90 mmol/l KCl, and it was abolished in 2K-1C arteries. The remaining response to acetylcholine in 2K arteries was blocked by L-NOARG. Thus, in addition to NO, a relaxing factor sensitive to extracellular K+ changes in the membrane potential contributes to endothelium-dependent relaxation in 2K-1C rat carotid artery. On the other hand, in arteries from 2K rats, only NO is involved in the relaxation induced by acetylcholine. The combination of 1H-[1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one (ODQ, 3 micromol/l), indomethacin (3 micromol/l) and L-NOARG (100 micromol/l) reduced the relaxation induced by acetylcholine in arteries from 2K-1C rats contracted with phenylephrine. On the other hand, in 2K arteries, the relaxation induced by acetylcholine was abolished. The combination of ODQ and K+ channel blockers charybdotoxin (100 nmol/l), apamin (500 nmol/l) and 4-aminopyridine (1 micromol/l) abolished the relaxation induced by acetylcholine in 2K and 2K-1C carotid arteries. These data indicate that the endothelium-derived relaxing factors that contribute to relaxation induced by acetylcholine are different in 2K and 2K-1C arteries. In 2K arteries, the only factor is NO, which involves the activation of K+ channels and the cGMP pathway. However, in 2K-1C arteries, the relaxation induced by acetylcholine is dependent on NO in addition to another factor, which is insensitive to indomethacin, but also activates the K+ channels and the cGMP pathway, presumably by membrane hyperpolarization through endothelium-derived hyperpolarizing factor.

Acetylcholine↗

PubMed2003

A membrane potential-sensitive dye for vascular smooth muscle cells assays.

Changes in membrane potential of rat aorta smooth muscle cells were investigated using the bis-oxonol sensitive probe DIBAC2(3). We compared the changes in membrane potential induced by a high external KCl concentration in aorta smooth muscle cells from normotensive 2 kidney (2K) and from renal hypertensive 2 kidney-1 clip (2K-1C) rats. The spectral properties of the membrane potential were first characterized in aqueous buffers and in cultured smooth muscle cells from 2K and 2K-1C rat aortas. Fluorescence emission and the images were recorded using a laser scanning confocal microscope. The relationship between fluorescence intensity (FI) and membrane potential (psi(m)) as a function of the increasing extracellular KCl concentration was linear in the 5-40 mmol/L KCl range in both 2K and 2K-1C rat aorta cells. Cell membranes from 2K-1C rat aorta cells were more depolarized (-55 mV) than 2K rat aorta cells (-65 mV). The results show that in 2K-1C aorta cells only 10 mmol/L KCl was needed to induce complete membrane depolarization while in 2K cells 40 mmol/L KCl was needed to induce a similar effect. This study clearly shows that the method is suitable to measure the membrane potential in cultured smooth muscle cells.

Animals↗

PubMedDate not supplied

Decreased vasodilation induced by a new nitric oxide donor in two kidney, one clip hypertensive rats is due to impaired k channel activation.

1. We studied the effect of the new compound trans-[RuCl([15]aneN(4))NO](2+) (15-ane) in denuded aortic rings of two kidney (2K) normotensive and two kidney, one clip (2K-1C) hypertensive rats. 2. The compound 15-ane releases nitric oxide (NO) when reduced by a catecholamine (noradrenaline). 3. Oxyhemoglobin (HbO(2)), an NO scavenger, completely abolished the effect of 15-ane in both 2K and 2K-1C rats, indicating that the relaxation is really due to NO release. 4. We tested the hypothesis that an impairment of K(+) channels plays an important role in the vasodilation induced by 15-ane. 5. The selective inhibitor of soluble guanylyl-cyclase, namely 1H-[1,2,4]oxadiazolo[4,3-alpha]quinoxalin-1-one (ODQ; 1 micromol/L) reduced the relaxation induced by 15-ane. In 2K-1C rat aortic rings, ODQ reduced the maximum effect (E(max)) of 15-ane, whereas in 2K rat aortic rings ODQ reduced E(max) and pD(2) values to 15-ane. 6. The selective K(+) channel blockers glibenclamide (blocks K(ATP); 3 micromol/L), 4-aminopyridine (blocks K(V); 1 mmol/L) and the small conductance K(Ca) channel blocker apamin (1 micromol/L) reduced E(max) and pD(2) values for 15-ane-induced relaxation responses of aortas from 2K rats. However, iberiotoxin, a blocker of large conductance K(Ca) channels, reduced only the E(max) to 15-ane. None of these K(+) channel blockers had any effect on the relaxation induced by 15-ane of aortas from 2K-1C rats. 7. These data indicate that an impaired functional activity of K(+) channels contributes to the deficient relaxation induced by the NO donor 15-ane in renal hypertensive 2K-1C rat aortas.

Animals↗