间歇性气囊挤压大鼠腿部对挤压部位和远端骨骼肌一氧化氮合酶mRNA表达的影响

目的:为了进一步了解间歇性气囊挤压法(Intermittent pneumatic compression, IPC)挤压大鼠腿部与一氧化氮(NO)的关系.方法:检测了大鼠骨骼肌中3种一氧化氮合酶(NOS)同工酶:神经型NOS(nNOS);诱导型NOS(iNOS)和内皮细胞型NOS(eNOS)mRNA在IPC作用后的表达变化.25只SD大鼠被随机分为3个模拟实验组和4个IPC实验组.每只鼠取右侧胫前肌(AT)和提睾肌(CM)作为正常对照.IPC组挤压0.5,1,和5h,及挤压5h加等待4h,模拟实验组除不挤压外,其他操作均与实验组相同,然后分离左侧AT和CM作为处理后样品.所有样品应用RT-PCR进行NOS mRNA测定.以样品中看家基因2,3-二羟基丙醛-3-磷酸脱氢酶(GAPHD)cDNA为内参,与NOS cDNA 共同扩增.PCR产物电泳条带密度用NIH图像分析软件定量,并以与正常对照的对比值作为变化比率.结果:在IPC作用0.5、1和5h后,eNOS mRNA显著上升,在AT中分别达到正常对照的1.2,1.8和2.6倍;在CM中分别达到1.2,1.8和2.7倍,而其他NOS,除5hIPC组的nNOS外,总体表现下调.在IPC作用1h加等待4h组中,eNOS mRNA回复至正常对照水平.结论:该结果证实了IPC产生的机械压力至少部分增加了血管壁的剪切压,使内皮细胞增加了NO产物的释放量,导致了挤压部位及远端肌肉的血管扩张和改善了微循环.     

NMDA受体与NOS在大鼠脊髓中间外侧柱的定位和生后发育

目的 探讨N-甲基-天冬氨酸受体（NMDAR1和NMDAR2A/B亚基）、一氧化氮合酶（NOS）及还原型辅酶Ⅱ硫辛酰胺脱氢酶（NADPH-d）活性在大鼠脊髓中间外侧柱（IML）的定位与生后发育特征。方法 在甲醛固定的脊髓切片上,进行ABC法免疫染色和NADPH-d组织化学反应与半定量分析。结果 MMDAR1,NMDAR2A/B,NOS I和NADPH-d反应产物丰富地分布于IML,主要定位于神经元胞体、树突和轴突样纤维终末。在生后早期发育中它们具有明显的动态变化,生后7d(P7）有微弱或中等的表达,随后逐渐上调,P21或P28达到高峰,然后保持于此水平于成体动物。结论 NMDA受体-NO通路可能是脊髓交感节前神经元一条重要的细胞内信号途径,并参与神经元生后发育成熟的调控过程。     

Phosphorus-Containing Fluoro-Sulfonated Polytriazole Membranes with High Proton Conductivity: Understanding Microstructural and Thermomechanical Behaviors as a Function of Degree of Sulfonation

The current article describes the design and synthesis of a new series of phosphorus-containing fluoro-sulfonated polytriazoles through click polymerization. The synthesized copolytriazoles (PTPFDSH-70 to 90) with different degrees of sulfonation (DS) from 70% to 90% are structurally interpreted by various spectroscopic techniques (¹H, ¹³C, and FTIR). The high molecular weight (weight average molecular weight as high as 77 500 g mol⁻¹ with polydispersity index of 2.29) polymers exhibits excellent mechanical (elongation at break up to 95%), thermal (10% decomposition temperature: 266–317 °C), and oxidative (>14.5 h) stability. The PTPFDSH-70 to 90 possess outstanding water-holding ability in hydrated conditions (swelling ratio [in-plane]: 6.2–7.3% at 80 °C). The microstructural alterations by their thermal relaxations and transitions with increasing DS in the polymers have been thoroughly investigated by dynamic mechanical analysis. The atomic force microscopy and transmission electron microscope images of the PTPFDSH-70 to 90 polymer membranes demonstrated the phase segregated interconnected ionic cluster-like morphology between hydrophilic and hydrophobic domains. The PTPFDSH-90 (DEB:PFAZ:DSAZ = 100:10:90) polymer membrane displays the proton conductivity (176 and 190 ms cm⁻¹ at 80 and 90 °C, respectively) higher than Nafion117 under similar test conditions.     

Cerebrovascular effects of nitric oxide manipulation in spontaneously hypertensive rats

1. Evidence that nitric oxide (NO) bioactivity is altered in chronic hypertension is conflicting, possibly as a result of heterogeneity in both the nature of the dysfunction and in the disease process itself. The brain is particularly vulnerable to the vascular complications of chronic hypertension, and the aim of this study was to assess whether differences in the cerebrovascular responsiveness to the NO synthase (NOS) inhibitors, N^G-nitro-L-arginine methyl ester (L-NAME) and 7-nitroindazole (7-NI), and to the NO donor 3-morpholinosydnonimine (SIN-1) might indicate one possible source of these complications.
2. Conscious spontaneously hypertensive (SHR) and WKY rats, were treated with L-NAME (30 mg kg⁻¹, i.v.), 7-NI (25 mg kg⁻¹, i.p.), SIN-1 (0.54 or 1.8 mg kg⁻¹ h⁻¹, continuous i.v. infusion) or saline (i.v.), 20 min before the measurement of local cerebral blood flow (LCBF) by the fully quantitative [¹⁴C]-iodoantipyrine autoradiographic technique.
3. With the exception of mean arterial blood pressure (MABP), there were no significant differences in physiological parameters between SHR and WKY rats within any of the treatment groups, or between treatment groups. L-NAME treatment increased MABP by 27% in WKY and 18% in SHR groups, whilst 7-NI had no significant effect in either group. Following the lower dose of SIN-1 infusion, MABP was decreased to a similar extent in both groups (around −20%). There was no significant difference in MABP between groups following the higher dose of SIN-1, but this represented a decrease of −41% in SHR and −21% in WKY rats.
4. With the exception of one brain region (nucleus accumbens), there were no significant differences in basal LCBF between WKY and SHR. L-NAME produced similar decreases in LCBF in both groups, ranging between −10 and −40%. The effect of 7-NI upon LCBF was more pronounced in the SHR (ranging from −34 to −57%) compared with the WKY (ranging from −14 to −43%), and in seven out of the thirteen brain areas examined there were significant differences in LCBF.
5. Following the lower dose of SIN-1, in the WKY 8 out of the 13 brain areas examined showed significant increases in blood flow compared to the saline treated animals. In contrast, only 2 brain areas showed significant increases in flow in the SHR. In the rest of the brain areas examined the effects of SIN-1 upon LCBF were less marked than in the WKY.
6. Infusion of the higher dose of SIN-1 resulted in further significant increases in LCBF in the WKY group (ranging between +30% and +74% compared to saline-treated animals), but no significant effects upon LCBF were found in the SHR. As a result, there were significant differences in LCBF between SIN-1-treated WKY and SHR in six brain areas. In most brain areas examined, cerebral blood flow in SHR following the higher dose of SIN-1 was less than that measured with the lower dose of SIN-1.
7. Despite comparable reductions in MABP (∼20%) in both groups, calculated cerebrovascular resistance (CVR) confirmed that the vasodilator effects of the lower dose of SIN-1 were significantly more pronounced throughout the brain in the WKY (ranging between −3% and −50%; median=−38%) when compared to the SHR (ranging between −10% and −36%; median=−26%). In the animals treated with the higher dose of SIN-1, CVR changes were broadly similar in both groups (median=−45% in WKY and −42% in SHR), but with the reduction in MABP in SHR being twice that found in WKY, this is in keeping with an attenuated blood flow response to SIN-1 in the SHR.
8. The results of this study indicate that NO-dependent vasodilator capacity is reduced in the cerebrovasculature of SHR. In addition, the equal responsiveness to a non-specific NOS inhibitor but an enhanced effectiveness of a specific neuronal NO inhibitor upon LCBF in the SHR could be consistent with an upregulation of the neuronal NO system.

     

Adrenergic,purinergic, and endothelial mediators and modulators of norepinephrine-induced mesenteric autoregulatory escape

We evaluated the effects of potential factors in autoregulatory escape from norepinephrine-induced vasoconstriction in rat anterior mesenteric artery. We determined mesenteric artery blood flow velocity with a pulsed Doppler, sonic flowmeter, and systemic arterial blood pressure with a transducer. A 4-min norepinephrine infusion (0.125–1.0 × 10^–8 M/min) intravenously evoked a dose-dependent, initial vasoconstriction that was followed by rapid escape of blood flow toward or above the control value during sustained norepinephrine administration. Neonatal capsaicin treatment enhanced vasoconstrictor responses to norepinephrine but failed to affect escape parameters. Propranolol decreased norepinephrine-induced escape dose dependently. Adenosine deaminase attenuated escape, and the combination of this enzyme plus propranolol nearly abolished escape from norepinephrine-induced vasoconstriction. Methylene blue also diminished autoregulatory escape. These findings suggest that norepinephrine-induced autoregulatory escape involves simultaneous -adrenoceptor, purinergic, and endothelial mediation. Norepinephrine-evoked mesenteric vasoconstriction appears to involve predominantly ₂-adrenoceptors and is modulated by peptidergic sensory nerves and adenosine.NIH grant number supporting these studies: USPHS # DK37050.     

The basal and stimulated release of the endothelium-derived relaxing factor from isolated pig coronary arteries does not interfere with the vascular release of superoxide

Oxygen-derived free radicals, in particular superoxide anions, are known to inactivate the endogenous vasodilator endothelium-derived relaxing factor (EDRF) which is probably identical with the gaseous radical nitric oxide. It is possible that EDRF is not the target of superoxide anions but may also be an endogenous scavenger of this radical.Superoxide anions generated by the vessel wall were measured by a modified lucigenin-enhanced chemiluminescence technique in isolated pig coronary artery rings with intact endothelium.The addition of bovine superoxide dismutase, a scavenger of superoxide anions, decreased the chemiluminescence signal by 40 ± 26% (mean ± SD; P < 0.05; n = 21) indicating reduced generation/release of superoxide anions. In contrast, pretreatment of coronary artery rings with diethyldithiocarbamate, an inhibitor of the intrinsic copper-zinc superoxide dismutase, increased the chemiluminescence response by 136 ± 128°10 (P < 0.05; n = 21). This increase in the chemiluminescence response induced by diethyldithiocarbamate-pretreatment was almost abolished in the presence of added bovine superoxide dismutase. Specific inhibition of the EDRF release with nitro-l-arginine (100 M) did not affect the chemiluminescence response. On the other hand, stimulation of the EDRF release by substance P (10 nM) or addition of the endothelium-mediated relaxant bradykinin (0.1 M) did not affect the chemiluminescence response. Stimulation of the EDRF release with serotonin (0.1 M) significantly reduced the photon emission by 15 ± 16% (n = 27). However, this effect of serotonin on the chemiluminescence response could not be prevented by specific inhibition of the EDRF release with nitro-l-arginine (100 M) but could be prevented by buffering the acidic serotonin solution with NaOH to pH 7.4.Our results suggest that basal and agonist-stimulated release of EDRF in isolated pig coronary artery rings does not interfere with the basal generation/release of superoxide anions derived from the vascular wall. Correspondence to: A. Mugge at the above address     

Evidence against a role of nitric oxide in the indirect negative inotropic-effect of M-cholinoceptor stimulation in human ventricular myocardium

Nitric oxide (NO) has been reported to mediate several effects in response to muscarinic cholinergic stimulation in cardiovascular tissues. Recently, an attenuation of guinea pig cardiac myocyte contraction by NO has been described. The aim of the present study was to determine whether the indirect negative inotropic effect of M-cholinoceptor stimulation in human myocardium is in part due to an effect of endogenous NO. Therefore, the effect of carbachol was studied under control conditions and during inhibition of NO-synthase by pretreatment with N^G-monomethyl-l-arginine (NMMA). Functional experiments were performed in isolated, electrically driven (1 Hz, 37°C) left ventricular papillary muscle strips of human myocardium. Since cytokines have been reported to be increased in the serum of patients with heart failure and could induce NO-synthase activity in failing myocardium, we compared samples from nonfailing and terminally failing (classified as NYHA IV) hearts. The indirect negative inotropic effect of carbachol (10 mol/l) was studied in the presence of the \-adrenoceptor agonist isoprenaline (0.03 mol/l).After stimulation with isoprenaline, carbachol significantly (P < 0.05) reduced force of contraction. This effect was diminished in failing myocardium compared to nonfailing, probably due to the diminished inotropic response most likely due to the lower cAMP levels in response to \-adrenoceptor stimulation in the former condition. Pretreatment with NMMA (100 mol/l) altered the antiadrenergic effect of carbachol neither in nonfailing nor in failing preparations. Furthermore, inhibition of guanylyl cyclase, the target enzyme of NO, by preincubation with methylene blue (10 mol/l) for 30 min had no effect on the carbachol-induced decrease in force of contraction. Basal force of contraction, as well as the positive inotropic effect of isoprenaline remained unaffected by NMMA or methylene blue.The present study provides evidence that the indirect negative inotropic effect of M-cholinoceptor agonists is not due to an effect of NO in the human myocardium. Furthermore, the well known enhancement of cGMP in response to M-cholinoceptor stimulation appears not to be involved in this antiadrenergic effect.     

Haematologic consequences of viral infections including serum iron status

Twenty-seven children with mumps or chickenpox were taken as a model to evaluate the haematological consequences of viral infections including serum iron status. Blood samples were obtained from all patients at presentation and on the 21 st day of the disease. While haemoglobin, haematocrit, and meam corpuscular volume levels were similar in two measurements (P>0.05), the mean leucocyte, absolute lymphocyte and thrombocyte counts, mean serum iron, serum iron binding capacity and transferrin saturation levels were lower at presentation than on the 21 st day of the disease. The serum iron levels were below 30 g/dl in 16 (59.2%) patients at presentation while only 4 (14.8%) had low values on 21 st day. Twenty-four (88.9%) patients had an increment in the serum iron binding capacity levels and 2 (7.6%) reached values above the normal range on the 21 st day. Int 21 (77.8%) patients, the transferrin saturation levels were below the expected ranges at presentation but 26 (96.3%) showed an increment on the 21 st day. However, the mean ferritin level was higher at presentation. Therefore, during the evaluation of patients for anaemia, the presence of a recent acute viral infection should be documented so as to avoid unnecessary iron medication.     

Reactive oxygen species in NMDA receptor-mediated glutamate neurotoxicity

In search of endogenous protective substances that inhibit neurotoxic action of glutamate and nitric oxide (NO), we found that brain-derived neurotrophic factor (BDNF), acting on TrkB receptor tyrosine kinase, inhibited neurotoxicity induced by glutamate and NO donors in cultured cortical neurons. In co-cultures of the mesencephalon and striatum, projection of mesencephalic dopamine neurons to the striatum attenuated N-methyl-d-aspartate (NMDA)-induced cytotoxicity in dopamine neurons themselves. Growth factors such as neurotrophins, which the target cells in the striatum would synthesize and secrete, may offer the protection of dopamine neurons against glutamate neurotoxicity.     

Effects of ventilatory pattern on exhaled nitric oxide in mechanically ventilated rabbits

BACKGROUND: Nitric oxide [NOexp] is present in exhaled air in many species. During experiments on pressure-controlled inverse ratio ventilation (PCIRV) in rabbits, increased [NOexp] was observed during PCIRV. The present study was undertaken to clarify which component of PCIRV increased [NOexp]. METHODS: Three groups of six New Zealand White rabbits were anaesthetised and mechanically ventilated. Exhaled nitric oxide, lung mechanics and gas exchange were measured using an experimental protocol designed to assess the effects of variations in 1) flow profile, 2) inspiratory time and 3) time-weighted tidal volume. Ventilator settings used were volume and pressure control ventilation at I:E ratios of 1:2 and 4:1. RESULTS: Constant and decelerating flow gave comparable [NOexp] levels (20.0 +/- 6.4 vs. 21.9 +/- 7.7 ppb, n.s.) when time-weighted tidal volume was kept constant. Using conventional (I:E 1:2) or inverted (I:E 4:1) I:E ratios in combination with decelerating flow and constant time-weighted tidal volumes did not alter [NOexp] (26.0 +/- 3.6 vs. 24.0 +/- 5.8 ppb, n.s.). An increased time-weighted tidal volume produced by pressure control with an I:E ratio of 4:1 increased [NOexp] (29.6 +/- 7.4) in comparison to constant (19.3 +/- 4.1, P < 0.05) and decelerating flow ventilation (19.6 +/- 3.6, P < 0.05) with I:E ratios of 1:2. CONCLUSION: The exhaled NO concentration was affected by ventilator setting. Increased levels of [NOexp] were observed with increases in time-weighted tidal volume, whereas changes in flow pattern and inspiratory time did not seem to influence airway NO production or release.