Aging and endothelin-1 induced vascular contractions

Summary Contractions produced by endothelin-1 (0.3–30 nM) have been investigated in aorta, renal arteries and mesenteric arteries from 2- and 24-month-old Sprague-Dawley rats. In senescent rats the EC₅₀ values of endothelin-1 for aorta and renal artery were significantly increased (aorta: from 6.2 to 12 nM; renal artery: from 5.2 to 7.8 nM). For mesenteric artery the EC₅₀ value (4.3 nM) was unchanged by aging, whereas the maximal contractile response to endothelin-1 was enhanced (from 8.3 to 11.7 mN). In contrast, there was no significant age-related difference in the maximal endothelin-1 response of aorta and renal artery. The present data demonstrate a reduced sensitivity for aorta and renal artery and an enhanced maximal response to endothelin-1 in the mesenteric artery in senescent rats.     

High sensitivity of porcine cerebral arteries to endothelin

Summary The threshold concentration of endothelin to induce contractions in porcine cerebral arteries (anterior cerebral, Willis ring and basilar artery) was lower than those for coronary and renal arteries. The median effective concentrations (ED₅₀) of endothelin in cerebral arteries were also significantly lower than those for coronary and renal arteries. There was no significant difference in the sensitivity to endothelin among cerebral arteries, or between coronary and renal arteries. The maximal percentage of contractions induced by endothelin, as compared to that induced by 10^–1 M potassium chloride, was not significantly different between the arteries.     

Extraneuronal serotonin accumulation in peripheral arteries of the rat

Accumulations of serotonin (5-HT) and norepinephrine (NE) were compared in control and 6-hydroxydopamine (6-OHDA) pretreated rat aorta, mesenteric and tail arteries. The distribution of these amines was corrected by subtracting tissue uptake of tritiated sorbitol in the extracellular space. 5-HT greatly accumulated both in control and 6-OHDA pretreated arteries. In contrast, NE accumulation in mesenteric and tail arteries was substantially decreased after 6-OHDA treatment. In the aorta 6-OHDA pretreatment did not affect the accumulation of both amines. These findings suggest that 5-HT accumulation in these arteries is mainly extraneuronal, and NE mainly neuronal. Since the accumulation of 5-HT in the aorta was not influenced by pretreatment with 10 microM NE, the extraneuronal uptake mechanisms for 5-HT and NE appear to be different.     

An analysis of the effects of urethane on cardiovascular responsiveness to catecholamines in terms of its interference with Ca++ mobilization from both intra and extracellular pools

Urethane (1 X 10(-2) - 1 X 10(-1) M) reduced, in a concentration-dependent manner, both intra and extracellular Ca++ dependent noradrenaline-induced contractions of perfused rabbit ear artery as well as the tonic contractions produced by perfusion with high K+ solution. However, a quantitative analysis of the data indicated that for urethane concentrations similar to those found in plasma during anesthesia urethane antagonism is confined to noradrenaline-induced contractions which depend upon the mobilization of Ca++ from intracellular storage sites. In KCl-contracted arteries, urethane enhanced the relaxant effects of isoprenaline. - Urethane reduced the amplitude of contractions of spontaneously beating guinea-pig right atrium at concentrations which have only a limited effect on frequency. In addition, it decreased in a concentration-dependent manner the amplitude of isoprenaline-activated electrically driven, and K+ depolarized guinea-pig right ventricular strips. Urethane had no effect on the chrono and inotropic actions of isoprenaline on cardiac preparations. In in vivo experiments the chronotropic response to low doses of isoprenaline was significantly higher in urethane-treated as compared to unanesthetized rats. The higher dose of isoprenaline tested produced a significant fall in systolic blood pressure in urethane-anesthetized rats. A significant correlation exists between the chronotropic response to isoprenaline and resting heart rate values in urethane-anesthetized rats. These results indicate that urethane, at concentrations similar to those found in plasma during anesthesia selectively interferes with mobilization of Ca++ from intracellular storage sites. In addition, the interference of urethane anesthesia with the isoprenaline chronotropic effect 'in vivo' cannot be explained by a direct interference of urethane with beta-adrenoceptors at cardiac level.     

Dopamine-beta-hydroxylase activity in stroke-prone spontaneously hypertensive rats

Dopamine-beta-hydroxylase (DBH) activity was higher in the serum, the mesenteric artery and the cerebral cortex of 4-week-old stroke-prone spontaneously hypertensive rats (SHRSP), and lower in the nucleus tractus solitarii than it was in spontaneously hypertensive rats (SHR).     

Inhibition of interleukin-1 beta release from cultured human peripheral blood mononuclear cells by prednisolone

Prednisolone, a water-soluble glucocorticoid hormone, suppressed the secretion of interleukin-1 beta from human peripheral blood mononuclear cells in culture. The prednisolone-induced suppression of the monokine release was dose-related and the half maximal response was observed at 0.1 nM.     

Influence of cyclic nucleotides on protein synthesis in vascular smooth muscle.

The incorporation of leucine-14C into protein in bovine mesenteric arteries was augmented by cyclic GMP (10-3 M) and decreased by cyclic AMP (10-3 M). There was no effect of 5'AMP (10-3 M). The phosphodiesterase inhibiting drugs theophylline (10-3 M) and papaverine (5 x 10-5 g/ml) both decreased the leucine-14C incorporation.     

Hydrocortisone administration enhances vessel contractility in Koltushi low-, and Koltushi high-avoidance rats

The contractility of tail artery rings was investigated in rats psychogenetically selected for rapid (KHA) and slow (KLA) acquisition of an avoidance response in the shuttle box. The vessel contractility was greater in the KHA rats. Hydrocortisone administration enhanced vessel contractility in both strains.     

Chemical sympathectomy reveals pre- and postsynaptic effects of neuropeptide Y (NPY) in the cardiovascular system

Intravenous injection of neuropeptide Y (NPY) caused short-lasting dose-dependent pressor responses in anesthetized rats. NPY was equipotent with noradrenaline in producing proportional pressor effects. Chemical sympathectomy, following the administration of 100 mg/kg 6-hydroxydopamine (6-OHDA), significantly potentiated the systemic pressor effects elicited by NPY or noradrenaline. Pretreatment with 2 nmol NPY enhanced the noradrenaline-induced pressor response in control rats. NPY did not change the basal tension of isolated rat aortic strips but significantly potentiated the contractile activity induced by 16 nM noradrenaline. This effect of NPY was not observed in aortic strips from rats pretreated with 6-OHDA. The presence of pre- and postsynaptic sites of action for NPY in the cardiovascular system of the rat is discussed.     

Chemical sympathectomy reveals pre-and postsynaptic effects of neuropeptide Y (NPY) in the cardiovascular system

Summary Intravenous injection of neuropeptide Y (NPY) caused short-lasting dose-dependent pressor responses in anesthetized rats. NPY was equipotent with noradrenaline in producing proportional pressor effects. Chemical sympathectomy, following the administration of 100 mg/kg 6-hydroxydopamine (6-OHDA), significantly potentiated the systemic pressor effects elicited by NPY or noradrenaline. Pretreatment with 2 nmol NPY enhanced the noradrenaline-induced pressor response in control rats. NPY did not change the basal tension of isolated rat aortic strips but significantly potentiated the contractile activity induced by 16 nM noradrenaline. This effect of NPY was not observed in aortic strips from rats pretreated with 6-OHDA. The presence of pre-and postsynaptic sites of action for NPY in the cardiovascular system of the rat is discussed.     

Dopamine-β-hydroxylase activity in stroke-prone spontaneously hypertensive rats

Summary Dopamine--hydroxylase (DBH) activity was higher in the serum, the mesenteric artery and the cerebral cortex of 4-week-old stroke-prone spontaneously hypertensive rats (SHRSP), and lower in the nucleus tractus solitarii than it was in spontaneously hypertensive rats (SHR).The authors wish to thank Prof. Masao Sano (Nagoya University) for his aid in dissecting the nucleus tractus solitarii and the locus coeruleus.     

Differential expression of genes for uncoupling proteins 1, 2 and 3 in brown and white adipose tissue depots during rat development

The different expression patterns of genes for uncoupling proteins (UCPs) 1, 2 and 3 (ucp1, ucp2 and ucp3) were studied in interscapular brown adipose tissue (BAT) and in four white adipose tissue (WAT) depots (epididymal, inguinal, mesenteric and retroperitoneal) in male rats of different ages (18 days-12 months). UCP mRNA expression levels were determined by Northern blotting. In BAT, there were high levels of expression of UCP1 and UCP3 mRNA, but no detectable levels of UCP2 mRNA. Both ucp1 and ucp3 followed a similar expression pattern with age, with high levels in suckling rats which decreased to 50% or less in rats just under 2 months old, declining thereafter until 5 months and then recovering with age. However, an additional peak of expression was observed for ucp3 at the age of 3 months. In WAT, ucp1 expression was rare: occasional expression was found for UCP1 mRNA in the retroperitoneal depot in suckling rats and in the epididymal and inguinal depots in suckling and mature adult rats. ucp2 and ucp3 had different developmental expression patterns, but these were similar for each gene in the different depots studied. UCP3 mRNA was highly expressed in rats soon after birth, it decreased until 3 months, and increased thereafter, except for the mesenteric WAT where ucp3 expression decreased until 7 months before recovering. The fact that changes with age of both ucp1 and ucp3 expression have a similar profile in BAT, which is also similar to the ucp3 and also ucp1 profiles in some WAT depots, might reflect a common regulatory pattern for the expression of these genes, and also a common function. In contrast to ucp1 and ucp3, ucp2 had a peak of expression at about 2 months, and lower expression at 3 months, suggesting different regulation and probably a different role for this UCP.     

64层螺旋CT血管成像对肠系膜上动脉活体形态的显示及临床应用

目的探讨64层螺旋cT血管成像（CTA）对肠系膜上动脉（SMA）活体形态的显示及临床应用价值。方法对80例临床疑有腹部、盆腔疾病的患者行64层螺旋CTA扫描,然后利用容积再现（VR）、多平面重建（MPR）、最大密度投影（MIP）、反向MIP（I—MIP）和薄层MIP（TS．MIP）等技术进行后处理。80例患者分为正常组（30例）和异常组（50例）两组,对正常组采用多种重建技术显示SMA各级分支,并测量SMA的直径,观察SMA起始段的走行;在MPR像上测量SMA与腹主动脉（AA）的夹角（An）,在左肾静脉（LRV）平面测量SMA至AA的距离（M）,在左侧肾门旁测量LRV最大前后径（A）,在SMA与AA问测量LRV最小前后径（B）。对异常组采用多种重建技术显示SMA,分析SMA特点。结果VR、MIP、I-MIP像均可显示正常组SMA整体解剖形态,TS-MIP对小分支显示优于其他重建技术。不同重建方式对SMA直径的测量无显著差异（F=0．71,P〉0．05）。SMA起始段走行方向为右下走行90％,前下走行占6．7％,左下走行占3．3％。An平均为（61．06±22．79）°。M平均为（14．62±4．00）mm,A平均为（9．16±1．91）mm,B平均为（6．00±2．01）mm,A与B比值〉2者占20．0％,〉3者占3．3％。异常组中有5例小肠先天性旋转不良及2例小肠扭转,SMA轴位像主要表现“回旋征”,VR表现为“螺旋征”;1例SMA分支栓塞;消化道肿瘤19例,其中8例供血动脉为SMA分支;回盲部动静脉畸形1例;余22例肝脏、泌尿生殖系统病变SMA无明显异常。结论64层螺旋CTA能很好地显示SMA的活体形态特征,具有重要的临床应用价值。     

Functional similarities in the mechanical design of the aorta in lower vertebrates and mammals

The mechanical properties of the aorta from the toad Bufo marinus, the lizard Gekko gecko and the garter snake Thamnophis radix were compared to those of the rat, by inflation of vessel segments in vitro. The arteries of the lower vertebrates, like those of mammals, were compliant, highly resilient, and non-linearly elastic. The elastic modulus of the artery wall was similar in the lower vertebrates and mammals, at their respective mean physiological pressures. We conclude that the aorta in each of these animals is suitably designed to function effectively as an elastic pulse smoothing component in the circulation; differences in the pressure wave transmission characteristics of lower vertebrates and mammals do not result from dissimilarities in arterial elastic properties, but from substantial differences in heart rate of these two groups.     

Differential vascular effects of neuropeptide Y(NPY) selective receptor agonists

Neuropeptide Y (NPY) increases blood pressure either directly or indirectly by potentiating the effect of various vasoconstrictors. Only one (the Y₁-receptor) of two subtypes of receptors (Y₁ and Y₂) is thought to mediate the vascular smooth muscle contraction. To test this hypothesis we challenged isolated rat mesenteric arteries that had a functional endothelium with (1–36) NPY and with specific Y₁-receptor ([Leu³¹, Pro³⁴] NPY) and Y₂-receptor ([Ahx^5–24, -Glu²--Lys³⁰] NPY) agonists. The Y₁-receptor agonist elicited a contractile response similar to that of NPY, whereas the Y₂-receptor agonist had no effect on wall tension. We also found that the presence of a functional endothelium has no influence on the contractile response to NPY. From these data we conclude that the direct contractile effect of NPY in the mesenteric artery is mediated by stimulation of Y₁-receptors and is not endothelium-dependent.     

Vascular reactivity in chronic Goldblatt two kidney-one clip hypertensive rats

We studied the possible contribution of increased vascular reactivity in the chronic phase of Goldblatt two kidney-one clip hypertension. Vascular reactivity was evaluated in aortic strips from hypertensive rats (16 weeks after inducing hypertension) and age-matched control rats. The findings were: a) increased sensitivity to vasopressin in the aortic tissue of hypertensive rats, b) a similar response to angiotensin II, noradrenaline and KCl in hypertensive and control rats, and c) reduced maximal response to angiotensin II compared with other vasoconstrictors in both groups of rats. These results suggest a possible role for vasopressin in the chronic phase of this model of hypertension.     

Demonstration of dopaminergic receptors in human pituitary adenomas secreting prolactin]

3H Domperidone binding on cellular membranes from human prolactin adenomas demonstrates the presence of two dopaminergic binding sites. The mean value of the dissociation constant (Kd) for five adenomas is of 0.29 +/- 0.14 nM for the first site and of 4.19 +/- 1.56 nM for the second site. The maximal number of binding sites (Bmax) varies from one adenoma to another. The binding is completely displaced at 30 nM of tritiated Domperidone by apomorohine, a specific dopaminergic agonist.     

Dynamic insulin secretion from perifused rat pancreatic islets

Insulin secretion from isolated pancreatic islets of 8- to 12-day-old rats was investigated in a dynamic in vitro (perifusion) system. The aims of the study were (i) to describe a carefully controlled in vitro method to study the mechanism of insulin secretion and to analyse the effects and dynamic interactions of bioactive compounds on isolated rat pancreatic islets, (ii) to validate the method by comparing fundamental data on the functions of the islets obtained with this method to those collected with other techniques; and (iii) to find novel features of the control of insulin secretion. The method was carefully designed to maintain the functional capacity of the explanted cells. A functional standardization system was elaborated consisting of (i) analysis of the changes in the basal hormone secretion of the cells; (ii) evaluating responses to a standard, specific stimuli (50 mM glucose for 3 min); (iii) determining the alteration of the momentary size of the hormone pool with responses to KCl; and (iv) direct determination of the total intracellular hormone content from the extract of the column. The technique provides accurate quantitative data on the dynamic responses to biologically active compounds that act directly on the pancreatic islets. The islets maintained their full responsiveness for up to 7 days, and responses as close as in 1-min intervals could be distinguished. A linear dose-response relationship was found on the glucose-induced insulin release in case of 3-min stimulation with 4 and 500 mM of glucose (lin-log graph). Utilizing this method, we showed that no desensitization to glucose-induced insulin release can be observed if the responsiveness of the cells is properly maintained and the parameters of the stimulation are carefully designed. Exposure of the explanted islets to 10 μM acetylcholine or 30 mM arginine (Arg) induced a transitory elevation of insulin release similar in shape to that experienced after glucose stimulation. Norepinephrine (NE), dopamine (DA) and somatostatin (SS) did not induce any detectable alteration on the basal insulin secretion of the islets. However, 100 nM SS given together with 50 mM glucose, 30 mM Arg or 10 μM acetylcholine significantly reduced the insulin-releasing effect of these substances (by 75.5, 71.5 and 72.5%, respectively). At the same time, SS did not alter the insulin response of the islets to 100 mM elevation of K⁺ concentration. SS also inhibited glucose-induced insulin release in a dose-dependent way (ED₅₀ = 22 nM). A similar dose-dependent inhibitory effect on glucose-induced insulin release was found with NE (ED₅₀ = 89 nM) and DA (ED₅₀ = 2.2 μM). γ-Aminobutyric acid (GABA) did not influence insulin release under similar circumstances. Received 16 January 1998; received after revision 6 May 1998; accepted 8 May 1998     

Influence of common bile duct cannula size on maximal secretory rate of taurocholate in the rat

Summary The common bile duct of male Sprague-Dawley rats was cannulated with either PE 10 or PE 50 tubing. Maximal secretory rate of taurocholate averaged 389±67 (SD) and 657±115 nmoles·min^–1·g liver^–1 in the PE 10 and PE 50 group, respectively (p<0.005). Maximal bile secretory pressure was significantly higher in the PE 10 group (240±28 vs 174±8 mm H20; p<0.005). When the maximal secretory rate was exceeded, bile flow decreased in both groups but this was accompanied with a decrease in maximal bile secretory pressure in the PE 10 group only. Maximal secretory rate of bile salts is markedly influenced by experimental technique. Use of small caliber common bile duct cannulae leads to partial obstruction and decreases the apparent maximal secretory rate for taurocholate.Acknowledgments. J. Reichen was the recipient of a Faculty Development Award in Clinical Pharmacology from The Pharmaceutical Manufacturer's Association Foundation, and is the recipient of a Research Career Development Award (KO4 AM 1189) from the National Institutes of Health. Supported by National Institutes of Health grant RO1 AM 27597.