Biomedical subjects

G M Buga

Publications and source records attributed to G M Buga.

32 records · Page 2Linked to original sources

PubMed1990

Antagonistic modulatory roles of magnesium and calcium on release of endothelium-derived relaxing factor and smooth muscle tone.

The objective of this study was to elucidate the mechanisms associated with the reciprocal relation between magnesium and calcium on vascular smooth muscle tone in bovine pulmonary artery and vein. Rapid removal of magnesium from Krebs-bicarbonate medium used to bathe isolated rings of precontracted artery or vein caused transient endothelium- and calcium-dependent relaxation and cyclic GMP accumulation. Both responses were antagonized by oxyhemoglobin, methylene blue, or superoxide anion and were enhanced by superoxide dismutase. The transient relaxation was followed by sustained endothelium-independent contraction. Endothelium-denuded vascular rings contracted in response to extracellular magnesium depletion without alteration in cyclic GMP levels. The data suggest that vascular endothelium-derived nitric oxide is responsible for the calcium-dependent relaxation elicited by extracellular magnesium depletion. Indeed, in bioassay cascade studies, magnesium removal from the medium used to perfuse intact artery or vein enhanced the formation and/or release of an endothelium-derived relaxing factor by calcium-dependent mechanisms. In the absence of both extracellular magnesium and calcium, calcium readdition caused transient endothelium-dependent relaxation and cyclic GMP accumulation, and both responses were abolished by oxyhemoglobin or methylene blue. In the presence of magnesium, however, readdition of calcium to calcium-depleted medium caused only contractile responses. Addition of magnesium to calcium-containing medium consistently caused endothelium- and cyclic GMP-independent relaxation that was not altered by oxyhemoglobin or methylene blue. Thus, magnesium and calcium elicit reciprocal or mutually antagonistic effects at the levels of both endothelium-derived relaxing factor formation and/or release and smooth muscle contraction. This relation may be of physiological importance, and the possibility that a reduction in circulating magnesium levels could lead to calcium-mediated vasospasm may be of pathophysiological concern.

Animals↗

PubMed1989

L-arginine causes whereas L-argininosuccinic acid inhibits endothelium-dependent vascular smooth muscle relaxation.

This study examined the actions of L-arginine, a putative precursor of endothelium-derived nitric oxide, and arginine analogs on endothelium-dependent relaxation of isolated rings of bovine pulmonary artery. L-Arginine did not consistently relax arterial rings unless rings were first rendered refractory to endothelium-dependent relaxation by pretreatment with 1 microM A23187 for 45 min. L-Arginine-elicited relaxation was endothelium-dependent, antagonized by oxyhemoglobin or methylene blue, and unaffected by indomethacin. L-Argininosuccinic acid caused endothelium-dependent contractions and irreversible inhibition of endothelium-dependent but not nitroglycerin-elicited relaxation, which was not overcome by addition of L-arginine. Inhibition of endothelium-dependent relaxation by L-NG-monomethyl arginine, however, was reversible and overcome by L-arginine. Therefore, endothelium-dependent relaxants may cause arginine depletion in endothelial cells and endogenous argininosuccinic acid may modulate the biosynthesis of endothelium-derived nitric oxide from arginine.

Acetylcholine↗

PubMed1989

Endothelium-derived nitric oxide relaxes nonvascular smooth muscle.

A bioassay cascade superfusion procedure was used to compare and contrast the actions of arterial and venous endothelium-derived relaxing factor (EDRF) with authentic nitric oxide (NO) on several nonvascular smooth muscle preparations. EDRF was released from human umbilical vein or bovine pulmonary artery by A23187 and allowed to superfuse two nonvascular and one vascular precontracted smooth muscle strips arranged in a cascade. NO or S-nitroso-N-acetylpenicillamine was delivered by superfusion. Both EDRF and NO relaxed bovine trachea, although artery was 10 times more sensitive than trachea to either relaxant. Similarly, rabbit taenia coli and rat fundus relaxed in response to high concentrations of NO or large amounts of EDRF released from umbilical vein. Vascular and nonvascular relaxant responses to both EDRF and NO were inhibited by oxyhemoglobin, methylene blue or superoxide, and were enhanced by superoxide dismutase. Perfusion of pulmonary artery or umbilical vein with A23187 resulted in contraction of guinea pig ileum and relaxation of pulmonary artery, whereas NO relaxed both preparations. Oxyhemoglobin enhanced the contractile and abolished the relaxant responses. Thus, ileum is more sensitive to endothelium-derived contracting factor(s) than to EDRF. NO raised cyclic GMP levels in all smooth muscle preparations, but a greater fold increase was observed in artery than in nonvascular smooth muscle. EDRF released from human umbilical vein was identified chemically as NO or a nitroso compound, as was done previously for EDRF from bovine pulmonary artery and vein. These observations support the view that one EDRF from artery and vein is NO or a labile nitroso compound.

Animals↗

PubMed1989

Basic polyamino acids rich in arginine, lysine, or ornithine cause both enhancement of and refractoriness to formation of endothelium-derived nitric oxide in pulmonary artery and vein.

The objective of this study was to elucidate the mechanism by which polyamino acids containing L-arginine, L-lysine or L-ornithine cause endothelium-dependent relaxation of bovine intrapulmonary artery and vein. Basic but not acidic or neutral polypeptides ranging in average molecular weights from 17 to 225 kDa elicited time- and concentration-dependent relaxation and cyclic GMP accumulation in precontracted rings of artery and vein by endothelium-dependent mechanisms. Vascular responses were markedly inhibited by oxyhemoglobin, methylene blue, or potassium. The basic polyamino acids stimulated the formation and/or release of an endothelium-derived relaxing factor (EDRF) identified as nitric oxide (NO) in perfused segments of both artery and vein as assessed by bioassay. The polyamino acids and A23187 released a similar endothelium-derived NO (EDNO) from artery and vein, as assessed by the similar half-life (3-5 seconds), antagonism by superoxide anion or oxyhemoglobin, enhancement by superoxide dismutase, and lack of influence by indomethacin. The basic polyamino acids elicited potent relaxant responses with EC50 values ranging from 3 x 10(-9) to 2 x 10(-7) M, and a direct correlation was obtained between molecular weight and relaxation potency irrespective of the basic amino acid incorporated. Prolonged contact of arterial or venous rings with basic polyamino acids resulted in the rapid development of marked refractoriness to relaxation and cyclic GMP formation on addition of polyamino acid. Moreover, refractoriness developed to the vascular responses of other endothelium-dependent vasodilators but not to glyceryl trinitrate or isoproterenol. The mechanism of refractory responses was attributed to interference with EDNO formation and release as assessed by bioassay and chemical assay. The hypothesis is forwarded that the basic polyamino acids serve as partial substrates for the enzyme system that catalyzes the conversion of L-arginine to NO. Prolonged contact between substrate and enzyme results in enzyme desensitization and the development of refractoriness or a form of tolerance to vasodilators whose action is mediated by EDNO.

Amino Acids↗

PubMed1988

EDRF generation and release from perfused bovine pulmonary artery and vein.

A bioassay cascade superfusion procedure was utilized to study the release and effects of endothelium-derived relaxing factor (EDRF) from isolated bovine pulmonary artery and vein. Three precontracted, endothelium-denuded strips of pulmonary artery or vein mounted in series served as the detector of EDRF. Perfusion of artery with acetylcholine, bradykinin or A23187, in the presence of indomethacin, caused the release of EDRF (t1/2 = 3-5 s) which relaxed superfused strips of both artery and vein. Similarly, perfusion of vein with bradykinin or A23187, but not acetylcholine, caused the release of EDRF (t1/2 = 3-5 s) which relaxed superfused strips of both vein and artery. EDRF-elicited vascular smooth muscle relaxation was associated with cyclic GMP accumulation in the superfused strips. EDRF activity was increased and its duration prolonged by superoxide dismutase or low oxygen tension, whereas oxyhemoglobin and methylene blue were markedly inhibitory. The similar patterns of release, instability, and properties of EDRF from intact artery and vein suggest that arterial and venous EDRF are closely similar.

Animals↗

PubMed1988

Pharmacological evidence that endothelium-derived relaxing factor is nitric oxide: use of pyrogallol and superoxide dismutase to study endothelium-dependent and nitric oxide-elicited vascular smooth muscle relaxation.

The principal objective of this study was to elucidate the influence of superoxide anion on both endothelium-dependent arterial relaxation elicited by acetylcholine and endothelium-independent arterial relaxation produced by nitric oxide (NO). Pyrogallol was used to generate superoxide in the oxygenated bathing medium, and superoxide dismutase was used to scavenge superoxide. Pyrogallol caused endothelium-dependent contractions of bovine intrapulmonary arterial and venous smooth muscle after precontraction of muscle by phenylephrine. Acetylcholine- and NO-elicited arterial relaxations were promptly converted to marked contractions upon addition of pyrogallol. Moreover, pyrogallol markedly inhibited the development of arterial relaxant responses to acetylcholine and NO. However, isoproterenol- and glyceryl trinitrate-elicited arterial relaxations were unaffected by pyrogallol. Both pyrogallol and oxyhemoglobin enhanced arterial contractile responsiveness to phenylephrine in an endothelium-dependent manner, whereas indomethacin was without effect. Similarly, both pyrogallol and oxyhemoglobin inhibited acetylcholine- and NO-elicited arterial cyclic GMP accumulation, whereas indomethacin was without effect. Uncontracted arterial rings maintained under tension showed endothelium-dependent contraction and decreased cyclic GMP levels in response to oxyhemoglobin but not pyrogallol. Superoxide dismutase enhanced arterial relaxation and cyclic GMP accumulation in response to both acetylcholine and NO. Using a bioassay superfusion cascade system in which intact perfused artery was the source of endothelium-derived relaxing factor (EDRF) and three endothelium-denuded arterial strips mounted in series served as the detector of EDRF, superfusion of strips with pyrogallol blocked relaxation caused by perfusion of artery with acetylcholine. Superoxide dismutase enhance the relaxations produced by arterial perfusion with acetylcholine and prevented the effects of pyrogallol.

Animals↗

PubMed1988

Endothelium-derived relaxing factor and nitric oxide possess identical pharmacologic properties as relaxants of bovine arterial and venous smooth muscle.

The principal objective of this study was to compare and contrast the vascular smooth muscle-relaxing properties of endothelium-derived relaxing factor (EDRF) and nitric oxide (NO) in two different assay systems. In one system, precontracted rings of bovine intrapulmonary artery and vein and coronary artery were used to compare the relaxant effects of endothelium-dependent vasodilators, NO, S-nitroso-N-acetylpenicillamine, glyceryl trinitrate, prostacyclin and isoproterenol. In a second system, a bioassay superfusion cascade procedure was employed to compare the relaxant effects and biologic stability of EDRF and NO. Acetylcholine and bradykinin elicited concentration-dependent but transient relaxant responses in arterial and venous rings, respectively. NO and S-nitroso-N-acetylpenicillamine, which generates NO in solution, elicited similar transient relaxant responses in endothelium-denuded arterial and venous rings. Glyceryl trinitrate, however, produced sustained relaxations, as did isoproterenol and prostacyclin. Utilizing a bioassay superfusion cascade system in which intact perfused artery or vein was the source of EDRF and three endothelium-denuded arterial or venous strips mounted in series served as the detector of EDRF or NO, the relaxation profile and biologic stability of superfused EDRF were compared with those of superfused NO. Arterial or venous perfusion with acetylcholine or bradykinin, respectively, and superfusion of NO over the strips produced characteristic decremental relaxant responses in the three vascular strips and revealed the highly unstable nature of EDRF and NO (T1/2 = 3-5 sec for both). The relaxation profile of EDRF was indistinguishable from that of NO.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗

PubMed1987

Endothelium-derived relaxing factor produced and released from artery and vein is nitric oxide.

The objective of this study was to determine whether nitric oxide (NO) is responsible for the vascular smooth muscle relaxation elicited by endothelium-derived relaxing factor (EDRF). EDRF is an unstable humoral substance released from artery and vein that mediates the action of endothelium-dependent vasodilators. NO is an unstable endothelium-independent vasodilator that is released from vasodilator drugs such as nitroprusside and glyceryl trinitrate. We have repeatedly observed that the actions of NO on vascular smooth muscle closely resemble those of EDRF. In the present study the vascular effects of EDRF released from perfused bovine intrapulmonary artery and vein were compared with the effects of NO delivered by superfusion over endothelium-denuded arterial and venous strips arranged in a cascade. EDRF was indistinguishable from NO in that both were labile (t1/2 = 3-5 sec), inactivated by pyrogallol or superoxide anion, stabilized by superoxide dismutase, and inhibited by oxyhemoglobin or potassium. Both EDRF and NO produced comparable increases in cyclic GMP accumulation in artery and vein, and this cyclic GMP accumulation was inhibited by pyrogallol, oxyhemoglobin, potassium, and methylene blue. EDRF was identified chemically as NO, or a labile nitroso species, by two procedures. First, like NO, EDRF released from freshly isolated aortic endothelial cells reacted with hemoglobin to yield nitrosylhemoglobin. Second, EDRF and NO each similarly promoted the diazotization of sulfanilic acid and yielded the same reaction product after coupling with N-(1-naphthyl)-ethylenediamine. Thus, EDRF released from artery and vein possesses identical biological and chemical properties as NO.

Animals↗

PubMed1987

Mechanisms of endothelium-dependent vascular smooth muscle relaxation elicited by bradykinin and VIP.

The objective of this study was to elucidate the mechanisms by which bradykinin and vasoactive intestinal polypeptide (VIP) relax bovine intrapulmonary artery and bradykinin, but not VIP, relaxes intrapulmonary vein. Bradykinin and VIP elicited entirely endothelium-dependent relaxation of phenylephrine-precontracted arterial rings, and this was associated with arterial accumulation of both guanosine 3',5'-cyclic monophosphate (cGMP) and adenosine 3',5'-cyclic monophosphate (cAMP). Bradykinin, but not VIP, relaxed precontracted venous rings and increased cGMP, but not cAMP levels, by endothelium-dependent mechanisms. Neither arteries nor veins relaxed in response to substance P, thrombin, bombesin, arginine vasopressin, or angiotensin II. Methylene blue or indomethacin each partially antagonized, whereas both, when together, abolished arterial relaxant responses to bradykinin and VIP. Methylene blue or indomethacin, respectively, abolished arterial cGMP or cAMP accumulation elicited by bradykinin and VIP. Venous relaxation and cGMP accumulation elicited by bradykinin was abolished by methylene blue but was unaltered by indomethacin. Thus bradykinin and VIP relaxed bovine intrapulmonary artery by endothelium-dependent mechanisms involving the actions of cGMP and cAMP whose formation may be stimulated by endothelium-derived relaxing factor and prostacyclin, respectively. In contrast, bradykinin relaxed intrapulmonary vein by endothelium-dependent mechanisms involving only cGMP.

Animals↗

PubMed1987

Endothelium-derived relaxing factor from pulmonary artery and vein possesses pharmacologic and chemical properties identical to those of nitric oxide radical.

The objective of this study was to elucidate the close similarity in properties between endothelium-derived relaxing factor (EDRF) and nitric oxide radical (NO). Whenever possible, a comparison was also made between arterial and venous EDRF. In vascular relaxation experiments, acetylcholine and bradykinin were used as endothelium-dependent relaxants of isolated rings of bovine intrapulmonary artery and vein, respectively, and NO was used to relax endothelium-denuded rings. Oxyhemoglobin produced virtually identical concentration-dependent inhibitory effects on both endothelium-dependent and NO-elicited relaxation. Oxyhemoglobin and oxymyoglobin lowered cyclic guanosine monophosphate (cGMP) levels, increased tone in unrubbed artery and vein, and abolished the marked accumulation of vascular cGMP caused both by endothelium-dependent relaxants and by NO. The marked inhibitory effects of oxyhemoglobin on arterial and venous relaxant responses and cGMP accumulation as well as its contractile effects were abolished or reversed by carbon monoxide. These observations indicate that EDRF and NO possess identical properties in their interactions with oxyhemoproteins. Both EDRF from artery and vein and NO activated purified soluble guanylate cyclase by heme-dependent mechanisms, thereby revealing an additional similarity in heme interactions. Spectrophotometric analysis disclosed that the characteristic shift in the Soret peak for hemoglobin produced by NO was also produced by an endothelium-derived factor released from washed aortic endothelial cells by acetylcholine or A23187. Pyrogallol, via the action of superoxide anion, markedly inhibited the spectral shifts, relaxant effects, and cGMP accumulating actions produced by both EDRF and NO. Superoxide dismutase enhanced the relaxant and cGMP accumulating effects of both EDRF and NO. Thus, EDRF and NO are inactivated by superoxide in a closely similar manner. We conclude, therefore, that EDRF from artery and vein is either NO or a chemically related radical species.

Acetylcholine↗

PubMed1986

In vitro evidence that vasoactive intestinal peptide is a transmitter of neuro-vasodilation in the head of the cat.

Experiments have been undertaken to determine the nature of the atropine-resistant neurogenic dilation that can be demonstrated in vitro in cephalic arteries of the cat. Levels of vasoactive intestinal peptide (VIP) and substance P were measured in a number of arteries and related to the extent of the neurogenic dilation that can be elicited in vitro. There is no correlation between the tissue contents of the two peptides. A positive correlation was found between vasoactive intestinal peptide but not substance P content and neurogenic dilation. Vasoactive intestinal peptide but not substance P consistently caused a concentration-dependent dilation of cephalic arteries not subject to significant tachyphylaxis. Vasoactive intestinal peptide antiserum in concentrations that block the dilation to vasoactive intestinal peptide (10(-6) M) but not that due to papaverine, significantly reduced neurodilation of both atropinized and non-atropinized lingual arteries--the cephalic artery with the highest VIP content. These results suggest that vasoactive intestinal peptide and not substance P significantly contributes to the non-cholinergic neurogenic dilation observed in vitro in arterial segments from the head of the cat.

Animals↗

PubMed1984

Vasoactive intestinal polypeptide and acetylcholine are responsible for neurovasodilation in the head of the cat.

There is reason to believe that the dilator innervation to blood vessels of many of the tissues of the head may be part of a common outflow from the central nervous system. Part of the effect of its activation is due to the release of acetylcholine and evidence is presented that, in addition, vasoactive intestinal polypeptide (VIP) release also plays a role. The non-cholinergic part of the dilator response to nerve activity can in a series of blood vessels be correlated with the level of VIP in their wall and is selectively reduced by VIP antiserum. In addition, VIP has suitable characteristics as a vasodilator. Whereas substance P which is present in the walls of many arteries of the head, does not qualify as a putative dilator transmitter. Despite the fact that the physiological role of dilator innervation to the circulation as a whole is not understood, the powerful consequences of the activation of the system in particular blood vessels suggests that this innervation should be seriously considered when studying the regulation of peripheral resistance.

Acetylcholine↗

PubMed1982

Distribution of choline acetyltransferase in cerebral and extracerebral cranial arteries of the cat. Its relationship to neurogenic atropine-sensitive dilation.

Choline-acetyltransferase (ChAT) activity was surveyed in segments of cranial arteries--both cerebral and extracerebral--from the cat. High levels were found in pial arteries, both cerebral and cerebellar, and in the arteries to salivary glands, tongue, and nose. Intermediate levels were found in the external and internal maxillary arteries and many of their branches. Enzyme levels in the arteries supplying the head--common carotid, vertebral, and in several systemic arteries and veins and also the lingual vein--were probably not significant. Only those vessels that have higher ChAT contents show capacity for neurogenic vasodilation. The dilation of segments of a number of these arteries, the basilar, middle cerebral, lingual, and internal maxillary, is reduced significantly by atropine (5 X 10(-7) M). ChAT activity did not correlate with vessel norepinephrine content. The data may be interpreted as defining a functional vasodilator system to the head encompassing both cerebral and extracerebral arteries that depends in part on a functional cholinergic link involving a muscarinic receptor. It is separate from the adrenergic outflow. The tissues supplied by vasculature receiving this type of innervation are of ectodermal origin.

Animals↗

PubMed1982

Further evidence for a muscarinic component to the neural vasodilator innervation of cerebral and cranial extracerebral arteries of the cat.

Transmural electrical stimulation of segments of lingual and cerebral (basilar, middle and posterior cerebral) and also other cranial arteries of the cat results after a long latency in a dilator response. The response may be resolved into two components--an initial transient atropine-sensitive component and a slower more ponderous one that is atropine-resistant. The variability in pattern of dilation responses from segments of different vessels or even those from the same segment of different cats is considerable. Some responses are entirely atropine-sensitive and others atropine-resistant; however the vast majority show a dilation that can be considered to be made up of both components. The latencies of the atropine-sensitive and atropine-resistant components are not different. The effect of atropine on the lingual but not the cerebral arteries is frequency dependent, being proportionately greater at low than at high frequencies. In both vessels, the effect of atropine is independent of train length at 1 Hz. Physostigmine potentiates significantly the dilation of the lingual artery but not that of the cerebral arteries. The potentiation is reversed by atropine. The endogenous acetylcholine level was measured in a series of vessels. It can be correlated with the activity of choline acetyltransferase and the presence of neurogenic dilation. It is proposed that there are two transmitters released in parallel from nerve(s) in the walls of cerebral, lingual, and possibly, other cranial arteries to cause vasodilation. It seems that one of these is acetylcholine.

Acetylcholine↗