Absence Of Intact Endothelium Research Articles

Autoantibodies Isolated from Preeclamptic Patients Induce Endothelial Dysfunction via Interaction with the Angiotensin II AT1 Receptor

Complete understanding of the etiology underlying endothelial damage in preeclampsia (PE) remains deficient. Recent studies suggest that autoantibodies against angiotensin II AT1 receptors (AT1-AA) may affect vascular endothelial integrity. However, direct evidence demonstrating association between AT1-AA from preeclamptic patients and vascular endothelial injury is lacking. The current study determined the effects of AT1-AA isolated from preeclamptic patients (Pre-IgG) upon the endothelium and attempted to elucidate the underlying mechanisms of injury. Pre-IgG markedly induced dose-dependent vasoconstriction in aortic vascular rings, an effect blocked by AT1 receptor antagonist losartan. Pre-IgG-induced vasoconstriction was increased in the absence of intact endothelium (1.59±0.04g vs. 1.63±0.08g, P<0.05). Additionally, Pre-IgG incubation with human umbilical vein endothelial cells significantly increased lactate dehydrogenase release in a time-dependent manner (0.84±0.07 vs. 3.50±0.09, 24 vs. 72-h exposure group, P<0.01) and increased caspase-3 and -8 activities (peaking at 48h), but did not affect caspase-9 activity. Taken together, these results support the contribution of AT1-AA to endothelial cell injury and dysfunction in PE.

The Novel Potent Multidrug Resistance Inhibitors N,N-bis(cyclohexanol)amine Aryl Esters Are Devoid of Vascular Effects

The aim of this study was to investigate the effects of the four isomers (3a, 3b, 3c and 3d) of a novel multidrug resistance-reverting agent – 3,4,5-trimethoxybenzoic acid 4-(methyl-{4-[3-(3,4,5-trimethoxyphenyl)acryloyloxy]cyclohexyl}amino)cyclohexyl ester – on vascular functions in vitro. A comparison of their mechanical and electrophysiological actions in rat aorta rings and single rat tail artery myocytes, respectively, was performed. In rat aorta rings, 3a–d antagonized both 60 mmol/l K⁺- and phenylephrine-induced contraction in a concentration-dependent manner, with maximal relaxation values averaging 50% of controls, 3d being the most effective of the series. The vasorelaxing effect was similar either in presence or absence of intact endothelium. In rat tail artery myocytes, out of the four isomers, only 3a consistently inhibited Ba²⁺ current through Ca_v1.2 channels. Our results provide functional evidence that 3a–d are weak vasorelaxing agents, although at concentrations much higher than those effective for multidrug resistance reversion in cancer cells.

Estrogen receptor-mediated vascular responsiveness to nebivolol: a novel endothelium-related mechanism of therapeutic vasorelaxation.

Nebivolol is a highly selective and lipophilic beta1-adrenergic receptor antagonist with vasodilating characteristics attributed mainly to endothelial generation of nitric oxide (NO). Coincidently, rapid vascular vasodilating effects of estradiol are also widely reported and membrane-initiated signaling by estrogen receptor (ER), leading to generation of NO, parallels the vasodilating effects observed for nebivolol. Thus, we hypothesized that the NO-dependent vasodilating effect attributed to nebivolol may be partially mediated through its interaction with the membrane-associated form of ER. The objective of this study was to examine the ER-mediated/endothelium-dependent vascular responses to nebivolol and the specific binding properties of nebivolol to ER. In isolated rat aortic rings, the endothelium-dependent vasodilating effect of nebivolol was significantly blocked by the use of the potent ER antagonist, ICI 182,780. In contrast, in the absence of intact endothelium, nebivolol-induced vasorelaxation was not blocked by ICI 182,780, strongly suggesting that nebivolol-elicited vasorelaxation is partially dependent on ER-mediated pathways. Further, we examined the binding of nebivolol to ER (MCF-7 cells) by radioligand binding assay and revealed specific binding kinetics with an estimated DC50 of 5 x 10 M, coinciding with the approximate EC50 of nebivolol as a vasorelaxant. In conclusion, the endothelium-dependent vascular response to nebivolol is attributed partially to its interaction with ER.

Dual vasoactive effects of tolazoline on rabbit pulmonary arteries

To determine the vasoactive effects of tolazoline on isolated rabbit pulmonary arteries. Prospective, in vitro, randomized, controlled trial. Experimental laboratory in a university-affiliated hospital. New Zealand White Rabbits. The pulmonary artery rings were obtained via thoracotomy. Their vasoactive responses were assessed in the presence and absence of intact endothelium and with or without precontraction by norepinephrine (NE, 3 x 10(-6) M) or potassium chloride (KCl, 3 x 10(-2) M). Using a tissue bath preparation, cumulative concentration response curves of tolazoline were obtained at different concentrations (10(-9) to 10(-4) M) after a period of stabilization. Tolazoline caused vasoconstriction of isolated pulmonary arteries without any pretreatment. The magnitude of the constriction was dose related and reached 300 g/g wet tissue at a concentration of 10(-4) M. On KCl-precontracted pulmonary arteries, tolazoline caused significant dose-related vasoconstriction. On the NE-precontracted vessel rings, it elicited significant dose-dependent vasodilation up to 60% relaxation at 10(-5) M. All the above effects were endothelium independent. Tolazoline has dual endothelium-independent vasoactive effects, causing vasoconstriction on isolated rabbit pulmonary arteries, either untreated or precontracted with KCl, and vasodilation on those precontracted with NE. Tolazoline may act as a competitive alpha-adrenoceptor blocking agent.

Comparative Vasoactive Effects of Amrinone on Systemic and Pulmonary Arteries in Rabbits

Amrinone has been increasingly used in management of low cardiac output syndrome during anesthesia, particularly when associated with right heart failure and pulmonary hypertension. This in vitro study was performed to determine and compare the direct vasoactive effects of amrinone on isolated rabbit systemic and pulmonary arteries. Responses of aortic and pulmonary artery rings from New Zealand white rabbits were assessed in the presence and absence of intact endothelium and with or without precontraction by norepinephrine (NE, 3 x 10(-6)M) or potassium chloride (KCl, 3 x 10(-2)M). Using a tissue bath preparation, cumulative concentration response curves of amrinone were obtained at different concentrations after a period of stabilization. Amrinone caused a dose-related vasodilation of NE-precontracted aortic and pulmonary arteries. It elicited about 65% and 90% relaxation, respectively, at a concentration of 300 mumol/L. Amrinone also induced a dose-related vasodilation of KCl-precontracted aortic and pulmonary arteries but to a lesser degree. All these effects were endothelium independent. By comparison, amrinone caused more relaxation in both NE- and KCl-precontracted pulmonary artery than aortic rings. In conclusion, amrinone has significant endothelium-independent, direct vasodilatory effects on isolated rabbit systemic and pulmonary arteries, more pronounced in the latter, particularly NE-precontracted vessels. Amrinone may have some calcium channel-blocking effect.

Vasoactive Effects of Ketamine on Isolated Rabbit Pulmonary Arteries

Ketamine has been used in patients with congenital heart disease and pulmonary hypertension with hypothetical controversy. Its direct effect on pulmonary arteries has not yet been clearly characterized. This in vitro study was performed to determine the direct vasoactive effects of ketamine on isolated rabbit pulmonary arteries. Responses of pulmonary artery rings from New Zealand white rabbits were assessed in the presence and absence of intact endothelium and with or without precontraction by norepinephrine (NE, 3 x 10(-6)M) or potassium chloride (KCl, 3 x 10(-2)M). Using a preparatory tissue bath, cumulative concentration response curves of ketamine were obtained at different concentrations (0.03, 0.1, 0.3, 1, 3 mM) after a period of stabilization. Ketamine caused a dose-related vasodilation on KCl-precontracted pulmonary arteries. It elicited almost 100% relaxation at a concentration of 3 mM. Ketamine also induced a dose-related vasodilation on NE-precontracted pulmonary arteries at a lesser degree. All of the effects were endothelium independent. In conclusion, ketamine has strong endothelium-independent, direct vasodilatory effects on isolated rabbit pulmonary arteries. Ketamine may act through Ca++ channel-blocking effect as well as inhibition of Ca++ release from sarcoplasmic reticulum.

Dual Effect of Clonidine on Isolated Rabbit Pulmonary Arteries

Clonidine, a partially selective agonist for alpha 2-adrenoceptors, has been increasingly used in anesthesia. Its direct effect on pulmonary arteries has not yet been clearly characterized. This in vitro study was performed to determine the vasoactive effects of clonidine on isolated rabbit pulmonary arteries. Responses of pulmonary artery rings from New Zealand white rabbits were assessed in the presence and absence of intact endothelium and with or without precontraction by norepinephrine (NE, 3 x 10(-6) M) or potassium chloride (KCl, 3 x 10(-2) M). Using tissue bath preparation, cumulative concentration response curves of clonidine were obtained at different concentrations (10(-8), 10(-7), 10(-6), 10(-5), 10(-4) M) after a period of stabilization. Clonidine caused vasoconstriction of isolated pulmonary arteries without any pretreatment. The magnitude of the constriction was dose related at lower concentrations and reached maximum of 300 g/g wet tissue when above 10(-6) M. On KCl-precontracted pulmonary arteries, clonidine caused significant dose-related vasoconstriction. On the NE-precontracted vessel rings, it elicited significant dose-dependent vasodilation up to 80% relaxation at 10(-4) M. All the above effects were endothelium independent. In conclusion, clonidine has dual endothelium-independent vasoactive effects, causing vasoconstriction on isolated rabbit pulmonary arteries, either untreated or precontracted with KCl, and vasodilation on those precontracted with NE. Clonidine may act as a competitive alpha-adrenoceptor blocking agent.

Bidirectional electrical communication between smooth muscle and endothelial cells in the pig coronary artery

Using strips of the left descending branch of the pig coronary artery in vitro, we show that smooth muscle cells are hyperpolarized by isoproterenol, a beta-agonist, independently of the presence or absence of intact endothelium. This hyperpolarization is transmitted to the lining endothelial cells in intact coronary strips. On the contrary, the cultured endothelial cells, without contact with smooth muscles, are not hyperpolarized by the beta-agonist. This shows that, in addition to the hyperpolarizations that flow from the endothelium to the media in response to kinins, electrical signals are also transmitted in the opposite direction, from the smooth muscles to the lining endothelial cells. In an attempt to test whether electrical coupling through gap junctions is implicated in the transmission of hyperpolarizations between the endothelial and the smooth muscle cells, we used halothane, a gap junction uncoupler. We observed that halothane does not inhibit the transmission of kinin hyperpolarizations from the endothelium to the smooth muscles, whereas it inhibits the transmission of isoproterenol hyperpolarization in the reverse direction.

Propofol Produces Endothelium-Independent Vasodilation and May Act as a Ca2+Channel Blocker

The mechanism of vasodilation induced by propofol was investigated using isolated rat thoracic aortic rings. Aortic rings were precontracted with potassium chloride (KCl) (40 mM) or phenylephrine (PE) (3 x 10(-8) to 3 x 10(-7) M) in the presence and absence of intact endothelium. Propofol produced similar concentration-dependent relaxation in aortic rings with and without endothelium regardless of whether they were precontracted with KCl or PE. The relaxation response to propofol was significantly greater in KCl-contracted aortic rings than in PE-contracted aortic rings. The propofol concentration producing 50% relaxation from the contracted state (RC50) was lower in aortic rings contracted with KCl than with PE, both with (5 +/- 0.6 x 10(-5) M vs 8.3 +/- 5.7 x 10(-4) M, P < 0.001) and without intact endothelium (3.9 +/- 0.5 x 10(-5) M vs 7.2 +/- 3.8 x 10(-4) M, P < 0.001). Propofol inhibited the Ca(2+)-induced contractions of aortic rings exposed to Ca(2+)-free media and depolarized with KCl (40 mM, 100 mM) in a dose-dependent manner. These effects are similar to those produced by verapamil. Propofol (5 x 10(-5) M) had minimal effect on the intracellular Ca2+ release elicited by PE (10(-5) M). We conclude that vasodilation produced by propofol is not endothelium-dependent but is likely due to blockade of voltage-gated influx of extracellular Ca2+.

The influence of endothelium on the action of PGF2α, and some dihydropyridine-type calcium antagonists in porcine basilar arteries

Vascular endothelium modulates the effect of various vasoconstricting mediators as well as the affinity of dihydropyridine-type calcium entry blockers. To further investigate this influence, vasoconstriction by PGF2 alpha as opposed to KCl and the affinity of nitrendipine and some related 3-ester side-chain derivatives were determined in isolated porcine basilar arteries in the presence and in the absence of intact endothelium, as well as in the presence of methylene blue. Treatment with methylene blue or mechanical endothelial damage increased the contractile work of basilar arteries stimulated by PGF2 alpha and reduced the affinity of the dihydropyridines in such precontracted vessels. Both experimental conditions resulted in nearly the same effect. In addition, the degree of intact endothelium, as determined by substance-P-induced vasodilation, significantly correlated with the corresponding efficacy of all dihydropyridines examined. In contrast, KCl-mediated contractions remained unchanged. It is suggested that the endothelium (probably due to the production and release of endothelium-derived vasorelaxing factors, such as EDRF and/or prostacyclin) may attenuate PGF2 alpha-induced transmembrane calcium influx through receptor operated calcium channels, whereas potential operated calcium channels seems to be unaffected.

Effect of argon laser irradiation on rabbit aortic smooth muscle: evidence for endothelium independent contraction and relaxation.

To study the effect of argon laser irradiation on vascular smooth muscle reactivity, ring segments of rabbit thoracic aorta were mounted isometrically in Krebs bicarbonate buffer. Laser irradiation was performed via a 200-micron core optical fibre interfaced with an argon ion gas laser. Laser irradiation at powers greater than 1.0 watt (W) (n = 34) produced contraction in each case, regardless of the duration of exposure. Conversely, irradiation at powers less than 0.1 W (n = 41) consistently produced relaxation. When the power employed ranged from 0.1 to 1.0 W (n = 100), the typical response consisted of a combination of low amplitude contraction and relaxation. At each power level, the responses observed were independent of the presence or absence of intact endothelium, assessed functionally by the response to acetylcholine and anatomically by scanning electron microscopy. Recordings of tissue temperature profiles performed during continuous-wave laser irradiation suggest that the findings observed in this study may be explained by a combination of heat induced contraction and light induced relaxation. At powers greater than 1.0 W, heat generated by laser irradiation is predominant, causing contraction. At powers less than 0.1 W, light predominates over heat, causing relaxation. When the power range is intermediate between these values, the heterogeneous response recorded reflects the combination of light and heat. In all cases, contraction and relaxation are endothelium independent.