Flecainide blocks the stimulatory effect of veratridine on slowly adapting pulmonary stretch receptors in anaesthetized rabbits without changing lung mechanics

We examined the responses of slowly adapting pulmonary stretch receptors (PSRs), total lung resistance (R_L) and dynamic lung compliance (C_dyn) to administered veratridine before and after pretreatment with atropine or flecainide in anaesthetized, artificially ventilated rabbits with bilateral vagotomy. Administration of veratridine (10 and 30 μg kg^-1) caused vigorous stimulation of PSRs, resulting in a tonic discharge of receptors during both inflation and deflation, but did not significantly alter either R_L or C_dyn. The veratridine-induced PSR stimulation became more prominent, as the dose of this alkaloid was increased. Pretreatment with atropine (1 or 2 mg kg^-1) had no significant effect on the excitatory response of PSRs to veratridine. The veratridine induced PSR stimulation was inhibited by treatment with flecainide (1, 2 and 3 mg kg^-1), a sodium channel blocker, and this inhibition was dose-dependent. These results suggest that activation of PSRs following veratridine administration probably related to the increased influx of sodium ions into the receptive terminals but does not depend upon bronchoconstriction.     

Lack of involvement of μ1 opioid receptors in dermorphin-induced inhibition of hypoxic and hypercapnic ventilation in rat pups

The effects of dermorphin, a mu-selective opioid agonist, on respiratory responses to altered O(2) and CO(2) during postnatal development were investigated in conscious, unrestrained Wistar rats aged 2-21 days. Respiration was recorded by barometric plethysmography. Dermorphin (4 mg kg(-1)) was administered subcutaneously, and the ventilatory responses to hypoxia (11% O(2), 89% N(2)) in 2-21-day-old pups and hyperoxia (100% O(2)), and hypercapnia (8% CO(2), 92% O(2)) in 2-13-day-old pups were assessed in the presence and absence of the mu(1) receptor antagonist naloxonazine (10 mg kg(-1) s.c.) administered 1 day before testing. Six minutes of hypoxia increased ventilation in all age groups, largely via an increase in frequency. Dermorphin inhibited the ventilatory response to hypoxia, and this inhibition was insensitive to naloxonazine. After 5 min of hyperoxia, ventilation was the same as with air breathing except in the presence of dermorphin, when hyperoxic ventilation was depressed by a naloxonazine-insensitive decrease in frequency. Following this 5 min 100% O(2) exposure, pups were exposed to hypercapnia, and respiratory parameters were measured 5 min later. The ventilatory response to CO(2) was inhibited by dermorphin in a naloxonazine-insensitive manner. There was no evidence for endogenous mu(1) receptor modulation of the ventilatory responses to altered gases in rat pups of any age. Thus, mu opioid-induced inhibition of the hypoxic and hypercapnic responses in young rats does not occur via activation of mu(1) opioid receptors.     

Responses of C fiber afferents of the rabbit airways and lungs to changes in extra-vascular fluid volume

Effects of changes in extra-vascular fluid volume produced by pulmonary lymphatic obstruction and plasmapheresis on the activities of bronchial and pulmonary C fiber receptors and rapidly adapting receptors (RARs) were investigated in New Zealand White rabbits. In intact rabbits, pulmonary lymphatic obstruction either alone or in combination with plasmapheresis did not stimulate pulmonary C fiber receptors. Only the combined stimulus activated the bronchial C fiber receptors. Bronchial C fiber receptors were also stimulated by graded increases in left atrial pressure (+5 and +10 mmHg). In contrast, RARs were activated by lymphatic obstruction either alone or in combination with plasmapheresis. These procedures increase the extra-vascular fluid volume in the carina and bronchi but not in the lungs (alveoli). In rabbits with chronic pulmonary venous congestion secondary to mitral valve damage, bronchial C fiber receptors were not stimulated by these increments in left atrial pressure which were insufficient to increase the extra vascular fluid content of the airways. However, both pulmonary and bronchial C fiber receptors were stimulated when the left atrial pressure was raised to 25 mmHg in these animals to cause pulmonary edema.     

Role of aquaporin and sodium channel in pleural water movement

The role of the ENaC sodium channel and aquaporin-1 (AQP1) water channel on pleural fluid dynamics in mice was investigated. 0.25 ml of hypertonic or isosmolar fluid was infused into the pleural space in anesthetized wildtype and AQP1 null mice. Pleural fluid was sampled at specified times to quantify the osmolality and volume. The sodium channel activator terbutaline increased isosmolar fluid clearance by 90% while the sodium channel inhibitor amiloride decreased it by 15%, but had no effect on osmotically driven water transport. AQP1 deletion significantly decreased osmotic water transport in pleural space by twofold, but it had no effect on isosmolar fluid clearance. Pretreatment with dexamethasone increased pleural osmotic fluid entry by 25%, while intravenous injection of HgCl2 decreased osmotic pleural water movement by 43%. These results provided evidence for a role of a sodium channel in pleural fluid absorption; AQP1 plays a major role in osmotic liquid transport but it does not affect isosmolar fluid clearance.     

The role of aquaporin-1 (AQP1) expression in a murine model of lipopolysaccharide-induced acute lung injury

A murine model of lipopolysaccharide (LPS)-induced acute lung injury (ALI) was used to evaluate whether aquaporin-1 (AQP1) is involved in lung inflammation and lung edema formation. Swiss strain mice (n = 122) had LPS (5 mg/kg) instilled intratracheally (IT), and were then treated with either 0.9 % saline or dexamethasone (5 mg/kg/day). Mice were euthanized at 2 days and 7 days after treatment. Inflammatory cytokines (TNF-alpha, IL-6), protein concentration in bronchoalveolar lavage (BAL) fluid, lung wet-to-dry weight ratio, histology, immunohistochemistry, and AQP1 Western blot were performed. Lung wet-to-dry weight ratio and lung vascular permeability were also measured in the AQP1 knockout mice (n = 9) that received IT LPS (5 mg/kg) at 2 days. Intratracheal instillation of LPS produced a severe lung injury at 2 days, characterized by elevation of TNF-alpha, IL-6 in the BAL fluid, and by histological changes consistent with increased lung vascular permeability and neutrophil infiltration. AQP1-immunoreactivity in the pulmonary capillary endothelium was reduced at 2 days and 7 days. Administration of dexamethasone improved LPS-induced ALI and retained expression of AQP1. However, depletion of AQP1 did not affect lung edema formation, lung vascular permeability, or lung histology. The results suggest that although AQP1 expression is decreased after lung injury, depletion of AQP1 does not alter lung inflammation and lung edema induced by LPS.     

Effects of C fiber blockade on cardiorespiratory responses to laryngeal stimulation in concious lambs

The primary aim of the study was to explore cardiorespiratory reflexes originating from laryngeal C fiber endings in the neonatal period. Seventeen lambs were instrumented for recording glottal adductor and diaphragm EMG, heart rate, systemic arterial pressure and respiratory movements. C fiber blockade was induced in eight lambs by 30 mg/kg capsaicin, the remaining nine lambs serving as controls. Cardiorespiratory reflexes were induced in non-sedated lambs by flowing air, menthol or 13% CO2, or by injecting water or 50 microg capsaicin in the laryngeal inlet through an endoscope. Responses to all stimuli but capsaicin were similar between the two groups. While cardiorespiratory responses were induced by capsaicin in control lambs, the responses were significantly inhibited in lambs with C fiber blockade. We conclude that laryngeal C fiber endings are functional and responsible for laryngeal chemoreflexes in newborn lambs.     

Effect of tacrolimus on endotoxin-induced lung injury in sheep

Since tacrolimus (FK-506) is known to suppress the proliferation and generation of T cells and to inhibit the production of T cell derived cytokines, we examined the effect of FK-506 on endotoxin-induced lung injury. We administered FK-506 (0.1 mg/kg) intravenously before the infusion of endotoxin (1 microgram/kg) into conscious sheep. We measured pulmonary hemodynamics, lung fluid balance, circulating leukocyte count and arterial blood gas tensions. The increase in pulmonary arterial pressure was significantly attenuated by FK-506 during the late period (3-5 h after endotoxin). Arterial oxygen gas tension was significantly higher in the FK-506 treated sheep during this phase. However, no significant differences were observed in lung lymph balance and circulating leukocyte count between the endotoxin alone group and the FK-506 treated group. These findings suggest that FK-506 may improve gas exchange in acute lung injury although there is an increased pulmonary vascular leakage. It is probable that FK-506 may have a beneficial potential on endotoxin-induced lung injury in sheep.     

Effects of inactivation and stimulation of locus coeruleus on respiratory activity of neonatal rat

We examined the effects of pharmacological inactivation and electrical stimulation of the locus coeruleus (LC) on the respiratory rate (RR) in isolated brainstem-spinal cord preparations of the neonatal rat. The brainstem and spinal cord were isolated en bloc from neonatal (days 1-4) Sprague-Dawley rats and superfused with artificial cerebrospinal fluid (ACSF) equilibrated with a gas mixture containing 2% CO(2) (pH 7.8). Pharmacological inactivation of the bilateral LC by means of microinjection of tetrodotoxin or noradrenaline elicited a significant decrease in RR in preparations obtained from rats aged 3-4 days, but not in preparations of rats aged 1-2 days. Stimulation of the bilateral LC with a train of electrical pulses (25-50 microA, 30 ms, 0.5 Hz, 3-4 min) caused a marginal but significant increase in RR on days 3-4, but not on days 1-2. These results indicate that the LC exerts an excitatory effect on the medullary respiratory rhythm generator in an age-dependent manner.     

Effects of abdominal distension on breathing pattern and respiratory mechanics in rabbits

The effects of acute abdominal distension (AD) on the electromechanical efficiency (Eff) of the inspiratory muscles were investigated in anesthetized rabbits by recording the electrical activity (A), pressure (P) exerted by the diaphragm (di) and parasternal intercostal muscles (ic), and lung volume changes when an abdominal balloon was inflated to various degrees. Eff,ic increased with increasing AD both in supine and upright postures. In upright rabbits Eff,di increased for intermediate but decreased at higher levels of AD, whilst it decreased at all levels of AD in supine rabbits. Tidal volume (VT) response followed that of Eff,di. Tonic Aic and Adi and inspiratory prolongation were elicited by AD. The effects of these neural mechanisms, acting to limit end-expiratory lung volume and VT changes, were however small since vagotomy prevented tonic Adi and inspiratory prolongation and reduced tonic Aic, but changed lung volume responses to AD only little. Hence, reduced respiratory system compliance and changes in inspiratory muscle electromechanical efficiency dominate lung volume responses to acute AD.     

Effect of domperidone on ventilation and polycythemia after 5 weeks of chronic hypoxia in rats

Chronically hypoxic humans and some mammals have attenuated ventilatory responses, which have been associated with high dopamine level in carotid bodies. Alveolar hypoventilation and blunted ventilatory response have been recognized to be at the basis of Chronic Mountain Sickness by generating arterial hypoxemia and polycythemia. To investigate whether dopamine antagonism could decrease the hemoglobin concentration by stimulating resting ventilation (VE) and/or hypoxic ventilatory response, 18 chronically hypoxic rats (5 weeks, PB=433 Torr) were studied with and without domperidone treatment (a peripheral dopamine antagonist). Acute and prolonged treatment significantly increased poikilocapnic ventilatory response to hypoxia (RVE ml/min/kg=VE at 0.1 FI(O(2))-VE at 0.21 FI(O(2))), from 506+/-36 to 697+/-48; and from 394+/-37 to 660+/-81, respectively. In addition, Domperidone treatment decreased hemoglobin concentration from 21.6+/-0.29 to 18.9+/-0.19 (P<0.01) in rats chronically exposed to hypobaric hypoxia. Our study suggests that the stimulant effect of D(2)-R blockade on ventilatory response to hypoxia seems to compensate the low hypoxic peripheral chemosensitivity after chronic exposure and the latter in turn decrease hemoglobin concentration.     

Role of L-glutamate in the locus coeruleus of rats in hypoxia-induced hyperventilation and anapyrexia

Locus coeruleus (LC) is a noradrenergic nucleus in the pons which has been reported to play an inhibitory role in the ventilatory response to hypoxia. Since LC contains glutamatergic receptors and L-glutamate is known to participate in the ventilatory and thermoregulatory responses to hypoxia, the effects of kynurenic acid (KYN, a glutamatergic receptor antagonist) microinjected into the LC in the hypoxic hyperventilation and anapyrexia (a regulated drop in body temperature [Tb]) were examined. Ventilation (V) and Tb were measured before and after a microinjection of KYN (10 nmol/0.1 microl) into the LC, followed by hypoxia. Control rats received a saline injection. Under normoxia, KYN treatment did not affect V or Tb. Typical hypoxia-induced hyperventilation and anapyrexia were observed after saline injection. KYN injection caused an increase in the ventilatory response, acting on tidal volume (Vt), but did not affect the anapyrexic response to hypoxia. These data suggest that L-glutamate in the LC is an excitatory neurotransmitter that activates an inhibitory pathway to reduce the hypoxic ventilatory response, similarly to the data reported for rostral ventrolateral medulla (VLM). The role of L-glutamate into the LC and VLM opposes its effect on other nuclei such as the nucleus of the solitary tract and ventromedullary surface, where the neurotransmitter participates in an excitatory pathway of the ventilatory response.     

咪达普利对氯化铵所致急性肺损伤大鼠血气、MDA及AngⅡ、CD54蛋白表达的影响

 目的: 本研究通过复制氯化铵致大鼠肺损伤模型,研究咪达普利对肺损伤大鼠动脉血气、MDA、肺形态学和AngⅡ、CD54蛋白表达的影响。方法: 将30只雄性大鼠随机分为3组:空白对照组、肺损伤模型组和药物组。对照组大鼠腹腔注射生理盐水2 mL/kg;肺损伤模型组大鼠腹腔注射等量6%氯化铵;药物组大鼠在注射6%氯化铵后,每天1次灌胃咪达普利(3 mg/kg),连续灌胃7 d;在实验第7天,用2%的戊巴比妥钠麻醉,采集腹主动脉血和腹腔静脉血样本;肺组织经4%多聚甲醛灌注、固定、脱水、透明、浸蜡、包埋和切片。ELISA法测定血清TNF-α和IL-6水平,化学发光法测定血清MDA含量,HE染色观察肺部形态学变化,免疫组织化学染色法观察肺组织AngⅡ和CD54蛋白表达。结果: 与对照组相比,模型组PaCO₂明显升高(P < 0.05),血清TNF-α、IL-6和MDA浓度显著增加(P < 0.01),氯化铵腹腔注射的肺损伤模型组大鼠出现肺水肿、肺炎、肺泡壁瘀血、肺泡腔出血和肺泡壁增生等损伤,肺组织AngⅡ和CD54蛋白表达明显增高(P < 0.01);与模型组比较,药物组PaCO₂明显降低(P < 0.05),血清TNF-α、IL-6和MDA浓度以及肺组织AngⅡ、CD54蛋白表达显著降低(P<0.01)。结论: 咪达普利可显著改善氯化铵所致肺损伤大鼠血气,抑制脂质过氧化和血清炎症细胞因子,降低肺组织AngⅡ和CD54蛋白表达。     

不同剂量左旋多巴对偏侧帕金森病大鼠行为学及多巴胺D2受体表达数的影响

目的：研究不同剂量左旋多巴对偏侧帕金森病大鼠行为学及多巴胺D₂受体表达数的影响，并对其相应机制进行探讨。方法：采用6-羟多巴胺（6-OHDA）制备偏侧PD大鼠模型，用免疫组化染色法检测PD模型大鼠分别给予左旋多巴不同剂量（10 mg·kg^-1·d^-1,50 mg·kg^-1·d^-1，100 mg·kg^-1·d^-1）腹腔注射15 d前后纹状体部位的多巴胺D₂受体表达数、观察大鼠行为学改变。结果：成功PD模型大鼠30 min旋转圈数与损毁侧及其健侧的D₂受体表达数增加百分数呈线性相关（r=0.927，P<0.01），大剂量左旋多巴干预后PD大鼠的旋转圈数明显少于对照组（P<0.05），且损毁侧纹状体多巴胺D₂受体表达数明显少于对照组（P<0.01）。结论：大剂量左旋多巴干预能明显改善PD大鼠的旋转行为，并使纹状体多巴胺D₂受体明显下调。     

Expression of HIF-1alpha, VEGF and VEGF receptors in the carotid body of chronically hypoxic rat

We examined the protein expression and localization of HIF-1alpha, VEGF, VEGF receptors in the carotid body (CB) of rats breathing 10% inspired oxygen for up to 4 weeks. The immunoreactivity (IR) of HIF-1alpha was distributed numerously in the nuclei of glomus (type-I) and other cells since hypoxia for 1 day, but was faint and scattered in the normoxic CBs. Cytoplasmic staining of the VEGF was intense in glomus cells of the hypoxic but not the normoxic group. The IR levels of HIF-1alpha and VEGF reached plateau at 4 weeks, and the IRs of VEGFR-1 and VEGFR-2 were strongly positive in the hypoxic group. Yet, the expression of VEGFR-1-IR was mild, whereas the VEGFR-2-IR was intense in normoxic CBs, suggesting an upregulation of VEGFR-1 but not VEGFR-2 in hypoxia. Hence, HIF-1 may activate the expression of VEGF and VEGFR-1 in the CB and the expression of VEGF in the chemoreceptors may play a paracrine role in the vascular remodeling during chronic hypoxia.     

Differential effect of epidermal growth factor on serous and mucous cells in porcine airway submucosal gland

Using a patch-clamp technique, we found that the fresh porcine submucosal gland acinar cells contained two functionally distinct cell populations, i.e. physiologically relevant concentration of acetylcholine (ACh, 30 nM) induced two distinct patterns of electric response in tracheal gland acinar cells. One was characterized by an outstanding oscillatory Cl(-)-current activity, and the other was with poor Cl(-)-current response but with a comparable K(+)-current. We examined the effect of epidermal growth factor (EGF) on the ACh-induced electric responses in these cells. EGF affected only the latter (K(+)-prominent) cell type to potentiate significantly the ACh-induced K(+)-current. An immunohistochemistry revealed that the receptor for EGF was identified preferentially on the mucous, but not serous, cells. Genistein, one of the tyrosine-kinase inhibitors, abolished the augmentation effect of EGF on the ACh-induced current. Thus, we identified the serous cell with a Cl(-)-rich current in response to ACh and the mucous cell with a K(+)-dominant response. Moreover, EGF affected the mucous cells alone to potentiate the ACh-induced electric response. EGF may contribute to the pathophysiological alterations in chronic inflammatory airways both in morphological (mucous cell hypertrophy/hyperplasia) and functional (thick viscous hypersecretion) ways.     

Effects on the rabbit carotid body of stimulation by almitrine, natural high altitude, and experimental normobaric hypoxia

Rabbits were given intraperitoneal injections of almitrine in ascending doses for 5 weeks. They were compared with a control group and with a group of rabbits which had been exposed from birth to the natural hypobaric hypoxia found at Cerro de Pasco (433 m) in the Peruvian Andes. A further group of animals was placed in an experimental normobaric chamber for either 3 or 6 months to subject them to the same degree of hypoxia as that occurring in Cerro. The carotid bodies of the rabbits in all these groups were processed for light and electron microscopy, and examined both qualitatively and quantitatively. The carotid bodies in the group given almitrine showed no changes in their size or in the population of their glomic cells when compared with controls. In contrast, the carotid bodies of Peruvian rabbits were greatly enlarged with a disproportionate increase in the population of the light variant of chief cell. Rabbits from the hypoxic chamber also had enlarged carotid bodies but those killed after 3 months showed an increase in the dark variant of chief cell, whereas after 6 months this cell was reduced in number. There was also intense cytoplasmic vacuolation. Election microscopy confirmed these changes and revealed that dark cells had larger, more pleomorphic granules than the light variant. Vacuolation of the granules in light cells was most pronounced in Peruvian rabbits, but was uncommon in animals exposed to hypoxia for 3 months. We suggest that the dark cell responds to the early stages of hypoxia but later matures into the light variant of chief cell.     

中药抗炎6号对大鼠腹腔巨噬细胞产生PGE,溶菌酶及C3b受体表达的影响

本文报道了中药抗炎6号(antiinflammatio No. 6 AI6)对大鼠腹膜腔巨噬细胞(rat peritoneal macrophages RPMφ)产生、分泌前列腺素E(prostaglandin PGE)、溶菌酶(lysozyme, LZM)及膜上C3b受体表达的影响。体外培养的RPMφ经酵母多糖刺激后,PGE;LZM的产生明显增加。AI6能明显地抑制酵母多糖刺激的RPMφ产生PGE(P<0.01)。对于未经刺激的RPMφ,AI6能明显地的促进其LZM的产生(P<0.01)。无论在体内或体外作用时,AI6对RPMφ膜表面C3b受体的表达都有明显的促进作用(P<0.01)。这些结果表明,AI6能增强Mφ的某些功能,这可能是AI6具有抗感染作用的基础。     

Effects of chloride ion substitution on the frequency dependence and α-autoinhibition of [3H]noradrenaline secretion in guinea-pig vas deferens

Guinea-pig isolated vas deferens, in which the stores of noradrenaline (NA) had been labelled by preincubation with [³H]NA, was used to study the effects of substitution of chloride with other anions, on transmitter secretion evoked by electrical stimulation of postganglionic sympathetic nerves. Chloride was omitted from the Tyrode's solution and substituted with acetate, nitrate, methylsulphate, sulphate or HEPES. Ail substituents increased both the spontaneous and the stimulus-evoked fractional secretion of [³H]NA under control conditions. The effect was essentially the same at all frequencies of stimulation (1–8 Hz). In the presence of the at-adrenoceptor blocker phentolamine, which enhanced the secretion of [³H]NA in all media, the secretion was not further enhanced by chloride substitution. Chloride substitution reduced the depressing effect of exogenous NA on the secretory response. In conclusion, chloride ions may be required for full expression of a-adrenoceptor-mediated inhibition of [³H]NA secretion, but do not markedly influence facilitation by increased frequency of stimulation.,     

Effect of chronic activation of 5-HT3 receptors on 5-HT3, 5-HT1A and 5-HT2A receptors functional activity and expression of key genes of the brain serotonin system

Among serotonin (5-HT) receptors, the 5-HT₃ receptor is the only ligand-gated ion-channel. Little is known about the interaction between the 5-HT₃ receptor and other 5-HT receptors and influence of 5-HT₃ chronic activation on other 5-HT receptors and the expression of key genes of 5-HT system. Chronic activation of 5-HT₃ receptor with intracerebroventricularly administrated selective agonist 1-(3-chlorophenyl)biguanide hydrochloride (m-CPBG) (14 days, 40 nmol, i.c.v.) produced significant desensitization of 5-HT₃ and 5-HT_1A receptors. The hypothermic responses produced by acute administration of selective agonist of 5-HT₃ receptor (m-CPBG, 40 nmol, i.c.v.) or selective agonist of 5-HT_1A receptor (8-hydroxy-2-(di-n-propylamino)tetralin) (8-OH-DPAT, 1 mg/kg, i.p.) was significantly lower in m-CPBG treated mice compared with the mice of control groups. Chronic m-CPBG administration failed to induce any significant change in the 5-HT_2A receptor functional activity and in the expression of the gene encoding 5-HT_2A receptor. Chronic activation of 5-HT₃ receptor produced no considerable effect on the expression on 5-HT₃, 5-HT_1A, and 5-HT transporter (5-HTT) and tryptophan hydroxylase-2 (TPH-2) genes – the key genes of brain 5-HT system, in the midbrain, frontal cortex and hippocampus. In conclusion, chronic activation of ionotropic 5-HT₃ receptor produced significant desensitization of 5-HT₃ and postsynaptic 5-HT_1A receptors but caused no considerable changes in the expression of key genes of the brain 5-HT system.