Modelo experimental de isquemia: reperfusão intestinal por clampeamento de aorta abdominal em ratos Wistar

OBJETIVO: desenvolver um modelo experimental de isquemia global normotérmica transitória capaz de demonstrar os tempos de isquemia e reperfusão necessários para desenvolvimento de lesão de isquemia/reperfusão em intestinos delgados de ratos Wistar através clampeamento de aorta abdominal suprarrenal. MÉTODOS: Vinte ratos Wistar adultos machos, pesando entre 250 e 350g, foram distribuídos aleatoriamente em cinco grupos, com quatro ratos cada, e submetidos a tempos crescentes de isquemia (0 - 30 - 45 - 60 - 90 minutos). Dentro de cada grupo, à exceção do grupo controle, dois ratos foram submetidos à 60 minutos de reperfusão e dois à 90 minutos. Após os procedimentos, procedeu-se análise histológica através de medição de áreas de necrose. RESULTADOS: O grau de necrose intestinal variou de 15 a 54% (p=0,0004). Houve tendência de aumento progressivo no grau de lesão relacionado ao aumento no tempo de isquemia, contudo, os maiores graus de lesão foram observados nos menores tempos de reperfusão. A análise do coeficiente de variação de necrose entre os dez grupos de isquemia/reperfusão demonstrou diferença estatisticamente significante em 15 postos, sendo 13 relacionados ao grupo controle. CONCLUSÃO: O modelo foi capaz de demonstrar os tempos necessários para que ocorra lesão de isquemia/reperfusão intestinal através de clampeamento aórtico e poderá servir como base para facilitar o desenvolvimento de estudos voltados para a compreensão deste tipo de lesão.

Avaliação da Velocidade Média na Aorta Torácica Descendente em Fetos com Anemia

Objetivo: verificar se existe correlação significativa entre a velocidade média na dopplerfluxometria da artéria aorta torácica descendente e o grau de anemia fetal. Métodos: estudo prospectivo, transversal, no qual foram analisados 66 fetos de gestantes isoimunizadas, em que se realizou a cordocentese para a realização de transfusões intra-uterinas pela via intravascular (66,7%). Nos fetos que foram submetidos à transfusão intra-uterina pela via intraperitoneal, ou naqueles casos em que não houve necessidade de tratamento intra-uterino (33,3%), a determinação da concentração de hemoglobina do cordão foi realizada pela punção do cordão umbilical, no momento da interrupção da gestação. Neste grupo de fetos estudados, foi realizado exame dopplerfluxométrico da artéria aorta torácica descendente, sendo calculada a velocidade média de fluxo. Foi realizado estudo de associação entre as variáveis. Foram também calculados os valores de sensibilidade, especificidade, valores preditivos positivo e negativo. Resultados: observou-se correlação significativa e inversa entre a velocidade média na artéria aorta torácica descendente e o nível de hemoglobina fetal. A velocidade média na dopplerfluxometria da artéria aorta torácica descendente apresentou sensibilidade de 47,5% para anemia fetal moderada (Hg<10 g/dL), com o teste exato de Fisher apresentando valor de p<0,01, e de 54,5% para anemia fetal grave (Hg<7,0 g/dL), com um valor de p=0,01. Conclusões: houve associação significativa entre a velocidade média na aorta torácica descendente e o grau de diagnóstico de anemia fetal.

Anatomical and radiographic appearance of the capuchin monkey thoracic cavity (Cebus apella)

The capuchin monkey is widespread both north and south of the Legal Amazon and in the Brazilian cerrado. Ten clinically healthy capuchin monkeys were submitted to an anatomical and radiographic study of their thoracic cavities. The radiographic evaluation allowed the description of biometric values associated with the cardiac silhouette and thoracic structures. Application of the VHS (vertebral heart size) method showed positive correlation (P<0.05) with depth of the thoracic cavity, as well as between the body length of vertebrae T3, T4, T5 and T6 and the cardiac length and width. The lung fields showed a diffuse interstitial pattern, more visible in the caudal lung lobes and a bronchial pattern in the middle and cranial lung lobes. The radiographic examination allowed preliminary inferences to be made concerning the syntopy of the thoracic structures and modification of the pulmonary patterns and cardiac anatomy for the capuchin monkey.

Thoracic and heart biometrics of non-anesthetized agouti (Dasyprocta primnolopha Wagler, 1831) measured on radiographic images

The agouti is a species intensively hunted throughout the Amazon and the semi-arid regions of northeastern Brazil. Considering the current trend in conservation management of wild species, the aim of this study was to determine the morphometric reference to the heart of agouti raised in captivity, based on thoracic and cardiac measurements in these animals. Thirty adult agoutis, 1 to 3 years of age, without clinical signs of cardiac disease were selected. The animals were physically restrained and radiographies in laterolateral (LL) and ventrodorsal (VD) recumbence were produced. The following measures were taken: the apicobasilar length of the heart (at the most cranial height of the Carina region to the heart apex) (AB), maximum width of the heart perpendicular to AB (CD), heart inclination angle (AIC), trachea inclination angle (AIT), distance from the right heart wall (DPTd), distance from the left heart wall (DPTe) and vertical depth of the thorax, and the ventral face of the vertebral column to the dorsal border of the sternum at the level of the trachea bifurcation (H). The ratios between AB/CD, AB/H and CD/H were also analyzed. To calculate the vertebral heart scale (VHS), the AB and CD measurements were laid over the thoracic vertebra starting at T4. Radiographic evaluation showed values consistent with those reported in small animals and some wild and exotic species. The main biometric values in the chest cavity and heart of agouti are arranged as follows: (1) The ratios between AB/H ratio and CD/H were not sensitive for identifying heart increases (p>0.05), while the ratio AB/CD was more sensitive in this identification (p<0.05); (2) AIC: 21.2±6.4º (mean between male and famale); (3) AIT for males and females: 9.93±3.23° and 8.4±3.94°; (4) DPTd and DPTe for males: 0.97±0.40cm and 0.7±0.30cm; (5) DPTd and DPTe for females: 1.12±0.42cm and 01.02±0.43cm; (6) VHS for males and females: 7.75±0.48v e 7.61±0.34v; (7) The caudal vena cava (CVC) was visualized dorsal-cranially and located right of the midline. The data obtained allowed the acquisition of the first reference values for biometry of the heart of agoutis, contributing to better understanding of cardiac morphology and identification of cardiomyopathy in these animals.

Os ramos colaterais da aorta abdominal em jaguatirica (Leopardus pardalis)

A jaguatirica (Leopardus pardalis) é uma das espécies de felino silvestre que pouco foi investigada quanto a sua morfologia. Assim, o estudo objetivou detalhar a origem e distribuição dos ramos colaterais da aorta abdominal deste animal. Avaliou-se dois exemplares, sendo um macho e uma fêmea, jovens, provenientes de Paragominas-PA, doados ao Laboratório de Pesquisa Morfológica Animal (LaPMA) da Universidade Federal Rural da Amazônia (UFRA). O sistema arterial foi preenchido com látex pigmentado de vermelho e os cadáveres foram preservados com solução de formaldeído tamponado a 10%. A aorta abdominal do L. pardalis teve origem entre T12 e L1, sendo a artéria celíaca o primeiro ramo visceral no sentido crânio-caudal, resultando nas artérias hepática, gástrica esquerda e esplênica. A artéria mesentérica cranial surgiu como segundo ramo da aorta abdominal, originando as artérias jejunais. Na sequência localizamos artéria pancreáticoduodenal caudal, artérias ileais, artérias ileocólicas, artérias renais direita e esquerda, artérias adrenais direita e esquerda e artérias ováricas ou testiculares direita e esquerda. Parietalmente, a aorta abdominal originou em média seis ramos lombares, bem como a artéria frenicoabdominal, as artérias circunflexas ilíacas profundas e artérias ilíacas externa e interna. A aorta abdominal gerou ainda a artéria mesentérica caudal, a qual dividiu-se em artérias cólica esquerda e retal cranial. A artéria cólica esquerda seguiu cranialmente paralela ao cólon descendente irrigando-o, originando em média 18 ramos, e anastomosando-se com a artéria cólica média. A artéria retal cranial seguiu em direção caudal distribuindo oito ramos à porção final do cólon descendente e ao reto, e uniu-se com a artéria retal média. Por fim, a aorta abdominal emitiu como ramo terminal a artéria sacral mediana. A vascularização arterial abdominal desta espécie é bastante semelhante ao descrito em felinos domésticos e demais mamíferos, com diferenças quanto ao número de artérias jejunais e origem das artérias renais.

Role of endothelium in angiotensin II formation by the rat aorta and mesenteric arterial bed

We investigated the angiotensin II (Ang II)-generating system by analyzing the vasoconstrictor effect of Ang II, angiotensin I (Ang I), and tetradecapeptide (TDP) renin substrate in the absence and presence of inhibitors of the renin-angiotensin system in isolated rat aortic rings and mesenteric arterial beds with and without functional endothelium. Ang II, Ang I, and TDP elicited a dose-dependent vasoconstrictor effect in both vascular preparations that was completely blocked by the Ang II receptor antagonist saralasin (50 nM). The angiotensin converting enzyme (ACE) inhibitor captopril (36 µM) completely inhibited the vasoconstrictor effect elicited by Ang I and TDP in aortic rings without affecting that of Ang II. In contrast, captopril (36 µM) significantly reduced (80-90%) the response to bolus injection of Ang I, without affecting those to Ang II and TDP in mesenteric arteries. Mechanical removal of the endothelium greatly potentiated (70-95%) the vasoconstrictor response to Ang II, Ang I, and TDP in aortic rings while these responses were unaffected by the removal of the endothelium of mesenteric arteries with sodium deoxycholate infusion. In addition, endothelium disruption did not change the pattern of response elicited by these peptides in the presence of captopril. These findings indicate that the endothelium may not be essential for Ang II formation in rat mesenteric arteries and aorta, but it may modulate the response to Ang II. Although Ang II formation from Ang I is essentially dependent on ACE in both vessels, our results suggest the existence of an alternative pathway in the mesenteric arterial bed that may play an important role in Ang II generation from TDP in resistance but not in large vessels during ACE inhibition

Differential effects of isoproterenol on the activity of angiotensin-converting enzyme in the rat heart and aorta

The excessive stimulation of beta-adrenergic receptors in the heart induces myocardial hypertrophy. There are several experimental data suggesting that this hypertrophy may also depend, at least partially, on the increase of local production of angiotensin II secondary to the activation of the cardiac renin-angiotensin system. In this study we investigated the effects of isoproterenol on the activity of angiotensin-converting enzyme (ACE) in the heart and also in the aorta and plasma. Male Wistar rats weighing 250 to 305 g were treated with a dose of (±)-isoproterenol (0.3 mg kg-1 day-1, N = 8) sufficient to produce cardiac hypertrophy without deleterious effects on the pumping capacity of the heart. Control rats (N = 7) were treated with vehicle (corn oil). The animals were killed one week later. ACE activity was determined in vitro in the four cardiac chambers, aorta and plasma by a fluorimetric assay. A significant hypertrophy was observed in both ventricular chambers. ACE activity in the atria remained constant after isoproterenol treatment. There was a significant increase (P<0.05) of ACE activity in the right ventricle (6.9 ± 0.9 to 8.2 ± 0.6 nmol His-Leu g-1 min-1) and in the left ventricle (6.4 ± 1.1 to 8.9 ± 0.8 nmol His-Leu g-1 min-1). In the aorta, however, ACE activity decreased (P<0.01) after isoproterenol (41 ± 3 to 27 ± 2 nmol His-Leu g-1 min-1) while it remained unchanged in the plasma. These data suggest that ACE expression in the heart can be increased by stimulation of beta-adrenoceptors. However, this effect is not observed on other local renin-angiotensin systems, such as the aorta. Our data also suggest that the increased sympathetic discharge and the elevated plasma concentration of catecholamines may contribute to the upregulation of ACE expression in the heart after myocardial infarction and heart failure.

Gap junctions in isolated rat aorta: evidence for contractile responses that exhibit a differential dependence on intercellular communication

Connexin43 (Cx43) is a major gap junction protein present in the Fischer-344 rat aorta. Previous studies have identified conditions under which selective disruption of intercellular communication with heptanol caused a significant, readily reversible and time-dependent diminution in the magnitude of a1-adrenergic contractions in isolated rat aorta. These observations have indentified a significant role for gap junctions in modulating vascular smooth muscle tone. The goal of these steady-state studies was to utilize isolated rat aortic rings to further evaluate the contribution of intercellular junctions to contractions elicited by cellular activation in response to several other vascular spasmogens. The effects of heptanol were examined (0.2-2.0 mM) on equivalent submaximal (»75% of the phenylephrine maximum) aortic contractions elicited by 5-hydroxytryptamine (5-HT; 1-2 µM), prostaglandin F2a (PGF2a; 1 µM) and endothelin-1 (ET-1; 20 nM). Statistical analysis revealed that 200 µM and 500 µM heptanol diminished the maximal amplitude of the steady-state contractile responses for 5-HT from a control response of 75 ± 6% (N = 26 rings) to 57 ± 7% (N = 26 rings) and 34.9 ± 6% (N = 13 rings), respectively (P<0.05), and for PGF2a from a control response of 75 ± 10% (N = 16 rings) to 52 ± 8% (N = 19 rings) and 25.9 ± 6% (N = 18 rings), respectively (P<0.05). In contrast, 200 µM and 500 µM heptanol had no detectable effect on the magnitude of ET-1-induced contractile responses, which were 76 ± 5.0% for the control response (N = 38 rings), 59 ± 6.0% in the presence of 200 µM heptanol (N = 17 rings), and 70 ± 6.0% in the presence of 500 µM heptanol (N = 23 rings) (P<0.13). Increasing the heptanol concentration to 1 mM was associated with a significant decrease in the magnitude of the steady-state ET-1-induced contractile response to 32 ± 5% (21 rings; P<0.01); further increasing the heptanol concentration to 2 mM had no additional effect. In rat aorta then, junctional modulation of tissue contractility appears to be agonist-dependent.

Vasorelaxant effects of the potassium channel opener SR 47063 on the isolated human saphenous vein and rat aorta

The vasorelaxant effects of SR 47063 (4-(2-cyanimino-1,2-dihydropyrid-1-yl)-2,2-dimethyl-6-nitrochromene), a new K+-channel opener structurally related to levcromakalim, were examined in isolated human saphenous vein (HSV) and rat aorta (RA). HSV or RA rings were precontracted with either KCl or noradrenaline and cumulative relaxant concentration-response curves were obtained for SR 47063 (0.1 nM to 1 µM) in the presence or absence of 3 µM glibenclamide. SR 47063 potently relaxed HSV and RA precontracted with 20 mM (but not 60 mM) KCl or 10 µM noradrenaline in a concentration-dependent manner, showing slightly greater activity in the aorta. The potency of the effect of SR 47063 on HSV and RA was 12- and 58-fold greater, respectively, than that reported for the structurally related K+-channel opener levcromakalim. The vasorelaxant action of SR 47063 in both blood vessels was strongly inhibited by 3 µM glibenclamide, consistent with a mechanism of action involving ATP-dependent K+-channels.

Experimental myocardial hypertrophy induced by a minimally invasive ascending aorta coarctation

Ascending aorta coarctation was produced by a minimally invasive technique in rabbits. Animal mortality was 5%. Morphometric and hemodynamic parameters were evaluated. A parabiotically isolated heart model was used to assess the hemodynamic parameters. Left ventricular weight/body weight ratio and muscle area showed clear evidence of hypertrophy when compared to control. The hemodynamic changes in the isolated heart model suggested decreased diastolic and systolic function in the coarcted group. The present model produced hypertrophy with low mortality rates as a result of its less invasive nature.

The distribution of presumptive thoracic paraganglionic tissue in the common marmoset (Callithrix jacchus)

The aortic-pulmonary regions (APR) of seven adult marmosets (Callithrix jacchus) and the region of the right subclavian artery of a further three marmosets were diffusion-fixed with 10% buffered formol-saline solution. In both regions serial 5-µm sections were cut and stained by the Martius yellow, brilliant crystal scarlet and soluble blue method. Presumptive thoracic paraganglionic (PTP) tissue was only observed in the APR. PTP tissue was composed of small groups of cells that varied in size and number. The distribution of the groups of cells was extremely variable, so much so that it would be misleading to attempt to classify their position; they were not circumscribed by a connective tissue capsule, but were always related to the thoracic branches of the left vagus nerve. The cells lay in loose areolar tissue characteristic of this part of the mediastinum and received their blood supply from small adjacent connective tissue arterioles. Unlike the paraganglionic tissue found in the carotid body the cells in the thorax did not appear to have a profuse capillary blood supply. There was, however, a close cellular-neural relationship. The cells, 10-15 µm in diameter, were oval or rounded in appearance and possessed a central nucleus and clear cytoplasm. No evidence was found that these cells possessed a 'companion' cell reminiscent of the arrangement of type 1 and type 2 cells in the carotid body. In conclusion, we found evidence of presumed paraganglionic tissue in the APR of the marmoset which, however, did not show the characteristic histological features of the aortic body chemoreceptors that have been described in some non-primate mammals. A survey of the mediastina of other non-human primates is required to establish whether this finding is atypical for these animals.

Calcium antagonism and the vasorelaxation of the rat aorta induced by rotundifolone

The vasorelaxing activity of rotundifolone (ROT), a major constituent (63.5%) of the essential oil of Mentha x villosa, was tested in male Wistar rats (300-350 g). In isolated rat aortic rings, increasing ROT concentrations (0.3, 1, 10, 100, 300, and 500 µg/ml) inhibited the contractile effects of 1 µM phenylephrine and of 80 or 30 mM KCl (IC50 values, reported as means ± SEM = 184 ± 6, 185 ± 3 and 188 ± 19 µg/ml, N = 6, respectively). In aortic rings pre-contracted with 1 µM phenylephrine, the smooth muscle-relaxant activity of ROT was inhibited by removal of the vascular endothelium (IC50 value = 235 ± 7 µg/ml, N = 6). Furthermore, ROT inhibited (pD2 = 6.04, N = 6) the CaCl2-induced contraction in depolarizing medium in a concentration-dependent manner. In Ca2+-free solution, ROT inhibited 1 µM phenylephrine-induced contraction in a concentration-dependent manner and did not modify the phasic contractile response evoked by caffeine (20 mM). In conclusion, in the present study we have shown that ROT produces an endothelium-independent vasorelaxing effect in the rat aorta. The results further indicated that in the rat aorta ROT is able to induce vasorelaxation, at least in part, by inhibiting both: a) voltage-dependent Ca² channels, and b) intracellular Ca2+ release selectively due to inositol 1,4,5-triphosphate activation. Additional studies are required to elucidate the mechanisms underlying ROT-induced relaxation.

Functional expression of kinin B1 and B2 receptors in mouse abdominal aorta

Previous studies have shown that the vascular reactivity of the mouse aorta differs substantially from that of the rat aorta in response to several agonists such as angiotensin II, endothelin-1 and isoproterenol. However, no information is available about the agonists bradykinin (BK) and DesArg9BK (DBK). Our aim was to determine the potential expression of kinin B1 and B2 receptors in the abdominal mouse aorta isolated from C57BL/6 mice. Contraction and relaxation responses to BK and DBK were investigated using isometric recordings. The kinins were unable to induce relaxation but concentration-contraction response curves were obtained by applying increasing concentrations of the agonists BK and DBK. These effects were blocked by the antagonists Icatibant and R-715, respectively. The potency (pD2) calculated from the curves was 7.0 ± 0.1 for BK and 7.3 ± 0.2 for DBK. The efficacy was 51 ± 2% for BK and 30 ± 1% for DBK when compared to 1 µM norepinephrine. The concentration-dependent responses of BK and DBK were markedly inhibited by the arachidonic acid inhibitor indomethacin (1 µM), suggesting a mediation by the cyclooxygenase pathway. These contractile responses were not potentiated in the presence of the NOS inhibitor L-NAME (1 mM) or endothelium-denuded aorta, indicating that the NO pathway is not involved. We conclude that the mouse aorta constitutively contains B1 and B2 subtypes of kinin receptors and that stimulation with BK and DBK induces contractile effect mediated by endothelium-independent vasoconstrictor prostanoids.

Lipolysis of emulsion models of triglyceride-rich lipoproteins is altered in male patients with abdominal aorta aneurysm

Disorders of the lipid metabolism may play a role in the genesis of abdominal aorta aneurysm. The present study examined the intravascular catabolism of chylomicrons, the lipoproteins that carry the dietary lipids absorbed by the intestine in the circulation in patients with abdominal aorta aneurysm. Thirteen male patients (72 ± 5 years) with abdominal aorta aneurysm with normal plasma lipid profile and 13 healthy male control subjects (73 ± 5 years) participated in the study. The method of chylomicron-like emulsions was used to evaluate this metabolism. The emulsion labeled with 14C-cholesteryl oleate and ³H-triolein was injected intravenously in both groups. Blood samples were taken at regular intervals over 60 min to determine the decay curves. The fractional clearance rate (FCR) of the radioactive labels was calculated by compartmental analysis. The FCR of the emulsion with ³H-triolein was smaller in the aortic aneurysm patients than in controls (0.025 ± 0.017 vs 0.039 ± 0.019 min-1; P < 0.05), but the FCR of14C-cholesteryl oleate of both groups did not differ. In conclusion, as indicated by the triglyceride FCR, chylomicron lipolysis is diminished in male patients with aortic aneurysm, whereas the remnant removal which is traced by the cholesteryl oleate FCR is not altered. The results suggest that defects in the chylomicron metabolism may represent a risk factor for development of abdominal aortic aneurysm.

The methyl ester of rosuvastatin elicited an endothelium-independent and 3-hydroxy-3-methylglutaryl coenzyme A reductase-independent relaxant effect in rat aorta

The relaxant effect of the methyl ester of rosuvastatin was evaluated on aortic rings from male Wistar rats (250-300 g, 6 rats for each experimental group) with and without endothelium precontracted with 1.0 µM phenylephrine. The methyl ester presented a slightly greater potency than rosuvastatin in relaxing aortic rings, with log IC50 values of -6.88 and -6.07 M, respectively. Unlike rosuvastatin, the effect of its methyl ester was endothelium-independent. Pretreatment with 10 µM indomethacin did not inhibit, and pretreatment with 1 mM mevalonate only modestly inhibited the relaxant effect of the methyl ester. Nω-nitro-L-arginine methyl ester (L-NAME, 10 µM), the selective nitric oxide-2 (NO-2) inhibitor 1400 W (10 µM), tetraethylammonium (TEA, 10 mM), and cycloheximide (10 µM) partially inhibited the relaxant effect of the methyl ester on endothelium-denuded aortic rings. However, the combination of TEA plus either L-NAME or cycloheximide completely inhibited the relaxant effect. Inducible NO synthase (NOS-2) was only present in endothelium-denuded aortic rings, as demonstrated by immunoblot with methyl ester-treated rings. In conclusion, whereas rosuvastatin was associated with a relaxant effect dependent on endothelium and hydroxymethylglutaryl coenzyme A reductase in rat aorta, the methyl ester of rosuvastatin exhibited an endothelium-independent and only slightly hydroxymethylglutaryl coenzyme A reductase-dependent relaxant effect. Both NO produced by NOS-2 and K+ channels are involved in the relaxant effect of the methyl ester of rosuvastatin.