Intravenous Injection Of Capsaicin Research Articles

Increased expression of macrophage migration inhibitory factor in the nucleus of solitary tract attenuates the renovascular hypertension

We investigatedwhether hemorrhagic hypotension (HH) altered the sensitivity of vagal pulmonary C-fibers in this study. The fiber activity (FA) of single vagal pulmonary C-fiber was continuously recorded in anesthetized rats before, during, and after HH was induced by bleeding from the femoral arterial catheter into a blood reservoir and lowering the mean systemic arterial blood pressure (MSAP) to ~40 mmHg for 20 min. Our results showed: 1)AfterMSAP reached a steady state of HH, the peak FA response to intravenous injection of capsaicin was elevated by ~5 folds. The enhanced C-fiber sensitivity continued to increase during HH and sustained even after MSAP returned to baseline during the recovery, but slowly returned to control ~20 min later. 2) Responses of FA to intravenous injections of other chemical stimulants of pulmonary Cfibers (phenylbiguanide, lactic acid and adenosine) and a constantpressure lung hyperinflation were all significantly potentiated by HH. 3) Infusion of sodium bicarbonate alleviated the systemic acidosis during HH, and it also attenuated, but did not completely prevent the HHinduced C-fiber hypersensitivity. In conclusion, the pulmonary C-fiber sensitivity was elevated during HH, probably caused by the endogenous release of chemical substances (e.g., lactic acid) that were produced by tissue ischemia during HH. This enhanced C-fiber sensitivity may heighten the pulmonary protective reflexes mediated through these afferents (e.g., J reflex) during hemorrhage when the body is more susceptible to other hazardous insults and pathophysiological stresses. (Supported in part by NHLBI and DoD grants)

A synergistic effect of TRPV1 and TRPA1 activations in vagal pulmonary C fibers (712.16)

Recent studies suggest that transient receptor potential ankyrin type 1 (TRPA1) channels can be activated by a number of inflammatory mediators and are largely co‐expressed with TRPV1 in a subset of vagal bronchopulmonary C‐fibers. This study was carried out to determine whether there is an interaction between these two polymodal transducers, TRPV1 and TRPA1, upon simultaneous activation. Pulmonary chemoreflex responses were compared in anesthetized, spontaneously breathing rats when intravenous injections of capsaicin (Cap, a TRPV1 activator; 0.75 µg/kg) and allyl isothiocyanate (AITC, a TRPA1 activator; 0.8‐1.2 mg/kg) were administered individually first and then in combination at 20 min intervals. The apneic ratios triggered by Cap alone and AITC alone were 2.2 ± 0.6 and 2.0 ± 0.9, respectively; intravenous injection of the combined dose (Cap + AITC) elevated the apneic ratio to 12.8 ± 2.4 which is ~255% greater than the mathematical sum of TRPV1 and TRPA1 responses (P<0.05, n=6). The synergistic effect was also observed in the study of chemoreflex responses in anesthetized mice (~450% greater than the mathematical sum of TRPV1 and TRPA1 responses, P<0.05, n=6; 1.3 ± 0.2, 1.1 ± 0.0, and 12.3 ± 2.9 for Cap alone, AITC alone, and Cap + AITC, respectively). Furthermore, when both TRPV1 and TRPA1 were activated simultaneously, the C‐fiber responses, recorded by the single‐fiber recording technique, were markedly amplified: responses to Cap alone, AITC alone and Cap + AITC were 4.3 ± 0.5, 3.4 ± 0.8 and 27.2 ± 3.4 impulses/s, respectively (~468% greater than the mathematical sum of TRPV1 and TRPA1 responses, P<0.05, n=24). These results clearly demonstrated a positive interaction between TRPV1 and TRPA1 activations at the pulmonary C‐fiber endings. The underlying mechanism of this synergistic effect remains to be determined. (Supported by NIH grant HL96914 and DoD grant W81XWH‐10‐2‐0189).

Warm SPA-induced hyperthermia confers protection to rats against airway inflammation evoked by capsaicin and substance P

Solus par aqua (SPA) is a traditional health care therapy. Warm SPA may enhance immunity and cellular defense to protect body against diseases. The present study investigated whether the warm SPA could confer protection to neurogenic inflammation in rats. The rats were immersed in water where the body core temperatures were maintained at hyperthermia (41.5 °C) or normothermia (37 °C) for a period of 15 min. After SPA for 1 or 6 days, neurogenic inflammation was induced by intravenous injection of capsaicin (90 μg/kg) or substance P (SP; 3 μg/kg). The plasma leakage and arterial pressures in rats after neurogenic inflammation were monitored. The extent of capsaicin- or SP-induced plasma leakage and hypotension was significantly attenuated in rats on day 1 after SPA hyperthermia. However, such resistance to neurogenic inflammation was not found on day 6 after hyperthermia. Western blotting analysis showed that the expression of heat shock protein 72 (HSP 72) in the trachea on days 1 and 2 after hyperthermia was 9.61-fold and 6.66-fold, respectively, of that in normothermia. Afterwards, the hyperthermia-induced HSP 72 upregulation gradually declined in a time-dependent manner. Thus, SPA hyperthermia may protect rats against neurogenic inflammation through modulation of HSP expression.

Capsaicin-evoked bradycardia in anesthetized guinea pigs is mediated by endogenous tachykinins

The present study was done to characterize the effects of endogenous tachykinins on heart rate in urethane-anesthetized guinea pigs. Intravenous injection of capsaicin (32 nmol/kg) was used to evoke release of tachykinins and calcitonin gene-related peptide (CGRP) from cardiac sensory nerve fibers. Such injections caused a brief decrease in heart rate (−37±7 beats/min, n=6) that was followed by a more prolonged increase (+44±10 beats/min). Blood pressure was lowered by −11±2 mmHg. Bilateral vagotomy did not affect the chronotropic or depressor responses to capsaicin, but atropine (1 µmol/kg) nearly abolished the bradycardic response (−8±3 beats/min, n=7). Combined blockade of NK2 and NK3 receptors, with SR48968 and SR14801 respectively, also caused a significant reduction of capsaicin-evoked bradycardia (−14±3 beats/min, n=4) but did not affect bradycardia evoked by vagal nerve stimulation. Blockade of CGRP receptors eliminated capsaicin-evoked tachycardia and prolonged the capsaicin-evoked bradycardia. These findings suggest that capsaicin-evoked bradycardia in the anesthetized guinea pig is mediated by tachykinins that stimulate cardiac cholinergic neurons. This effect appears to be truncated by the positive chronotropic action of CGRP that is also released from cardiac afferents by capsaicin.

Prostaglandin involvement in lung C-fiber activation by substance P in guinea pigs

Airway hyperresponsiveness is a cardinal feature of asthma. Lung C-fiber activation induces central and local defense reflexes that may contribute to airway hyperresponsiveness. Initial studies show that substance P (SP) activates C fibers even though it is produced and released by these same C fibers. SP may induce release of other endogenous mediators. Bradykinin (BK) is an endogenous mediator that activates C fibers. The hypothesis was tested that SP activates C fibers via BK release. Guinea pigs were anesthetized, and C-fiber activity (FA), pulmonary insufflation pressure (PIP), heart rate, and arterial blood pressure were monitored before and after intravenous injection of capsaicin (Cap), SP, and BK. Identical agonist challenges were repeated after infusion of an antagonist cocktail of des-Arg9-[Leu8]-BK (10(-3) M, B1 antagonist), and HOE-140 (10(-4) M, B2 antagonist). After antagonist administration, BK increased neither PIP nor FA. Increases in neither PIP nor FA were attenuated after Cap or SP challenge. In a second series of experiments, Cap and SP were injected before and after infusion of indomethacin (1 mg/kg iv) to determine whether either agent activates C fibers through release of arachidonic acid metabolites. Indomethacin administration decreased the effect of SP challenge on FA but not PIP. The effect of Cap on FA or PIP was not altered by indomethacin. In subsequent experiments, C fibers were activated by prostaglandin E2 and F2alpha. Therefore, exogenously applied SP stimulates an indomethacin-sensitive pathway leading to C-fiber activation.

Local capsaicin application to the stellate ganglion and stellatectomy attenuate neurogenic inflammation in rat bronchi

The present study investigated the contributions of vagal and nonvagal sensory nerve fibers on neurogenic inflammation in rat bronchial airways. A surgical procedure was developed via the rat mediastinum ventral intercostal space to prepare an intercostal opening without causing pneumothorax for performing stellate ganglionectomy alone, thoracic vagus nerve section alone, and stellatectomy plus thoracic vagotomy, and for injecting capsaicin (2 μl, 10 mg/ml) and 6-hydroxydopamine (2 μl, 50 mg/ml) into the ganglion. One week later in our procedure, we investigated if neurogenic inflammation induced by an intravenous injection of capsaicin (300 nmol/ml/kg) and innervation density of substance P-immunoreactive sensory axons could be decreased after chronic denervation in the rat lower airways. The major findings were that surgical removal of the right stellate ganglion and local capsaicin application resulted in a significant attenuation of neurogenic plasma extravasation in the right bronchial tree evoked by systemic capsaicin application. Reduction of neurogenic plasma extravasation was totally abolished by combined stellatectomy and thoracic vagotomy. The number of substance P-containing axons was also greatly decreased following these surgical and capsaicin treatments. It is concluded that sensory nerve fibers from both vagal source and nonvagal (spinal) source, which associated with the stellate ganglion, contributed significantly to neurogenic inflammation in the bronchial airways with a slightly higher contribution from the vagus nerve.

CGRP, but not substance P, induces an increased negativity of the interstitial fluid pressure in rat trachea.

Neurogenic inflammation is mediated by neuropeptides released from sensory nerves following electrical stimulation of the vagal nerve or by capsaicin. The released neuropeptides are, among others, calcitonin gene-related peptide and substance P, which both induce vasodilation, while only substance P induces plasma extravasation. Electrical stimulation of the vagal nerve induces increased negativity of interstitial fluid pressure (Pif), which will contribute to enhance oedema formation. Pif was measured, on the abluminal side of the surgically exposed trachea, with sharpened glass capillaries (4-10 microns) connected to a servo-controlled counterpressure system. Measurements were performed after circulatory arrest, since the oedema formation associated with acute inflammation will increase Pif in a positive direction, which may potentially underestimate the increased negativity of Pif. Experiments were carried out in pentobarbital anaesthetized (50 mg kg-1) Wistar-Møller rats. Pif in control rats averaged -1.2 +/- 0.9 (SD) mmHg (n = 9). Intravenous injection of capsaicin (65.0 nmol) and calcitonin gene-related peptide (1.3 nmol) increased the negativity of Pif to -4.0 +/- 1.2 mmHg (n = 8) (P < 0.01) and -4.7 +/- 2.0 mmHg (n = 9) (P < 0.01), respectively. Intravenous injection of substance P (7.4 nmol, n = 9; and 37.0 nmol, n = 8) did not affect Pif compared to control (P > 0.05). Similarly, potentiation of the available substance P with thiorphan or captopril did not increase the negative Pif, nor did injection of stable substance P analogues. Thus, the present study seems to support the theory that, in rat trachea, the increased negativity of Pif after intravenous injection of capsaicin and after vagal stimulation is caused by calcitonin gene-related peptide.

Role of endothelin in mediating neurogenic plasma extravasation in rat dura mater

In addition to their potent vasoconstrictor properties, the endothelins (endothelin-1 and -3) may possess neurotransmitter/neuromediator and neuroendocrine actions. The aim of the present study was to evaluate the role of endothelins (ET) in mediating neurogenic inflammation of cephalic tissues in the rat. For this purpose, bosentan, a specific non-peptide mixed antagonist of ET receptors, was tested in rat models of neurogenic and non-neurogenic plasma extravasation in the dura mater and extracranial tissues (eyelid, conjunctiva, lip, tongue). Bosentan was effective for preventing neurogenic inflammation in the dura mater induced by unilateral electrical stimulation of the trigeminal ganglion or intravenous injection of capsaicin, whereas it was ineffective in extracranial tissues or after injection of substance P (non-neurogenic inflammation). The effect of nerve fiber stimulation on ET plasma concentrations in superior sagittal sinus was measured using selective radioimmunoassays for ET-1 and -3. Endothelin-3 concentration significantly increased after intravenous injection of capsaicin, whereas ET-1 levels remained unchanged. Competition binding assays on microsomal membranes from the trigeminal ganglion revealed a single class of binding sites with equal affinity for ET-1 and ET-3, suggesting a homogenous population of ET B receptors. The role of ET B receptors in mediating inflammation was evidenced by the lack of efficacy of a selective ET A receptor antagonist, in contrast to the full efficacy of a selective ET B receptor antagonist, for preventing neurogenic inflammation induced by unilateral stimulation of the trigeminal ganglion. The role of ET B receptors was finally confirmed by the observation that exogenous administration of the ET B receptor agonist sarafotoxin S6c also induced plasma protein extravasation in the dura mater. This extravasation was not a direct effect of ET B receptor stimulation, because it was inhibited by spantide, a selective tachykinin receptor antagonist. These data strongly suggest that ET, acting through ET B receptors, may play an important role in mediating neurogenic inflammation in the meninges of rats. Since the profile of activity of bosentan is similar to that of the 5- HT 1 D B agonists, sumatriptan and ergot alkaloios, one may speculate that ET receptor antagonists might be potentially effective in the treatment of acute migraine attacks.

Inhibition of neurogenic inflammation in the meninges by a non-peptide NK 1 receptor antagonist, RP 67580

RP 67580, a non-peptide NK 1 receptor antagonist, inhibited in a stereoselective and dose-dependent manner plasma extravasation caused in the dura mater by intravenous injection of capsaicin in guinea-pigs and of exogenous substance P in rats (ED 50 = 35 and 2.5 μg/kg i.v., respectively). In the two species, RP 67580 appeared to be more effective in the dura mater than in the peripheral organs. These results indicate that selective NK 1 receptor antagonists could be potentially effective for the treatment of migraine headache.

Mycoplasma pulmonis infections cause long-lasting potentiation of neurogenic inflammation in the respiratory tract of the rat.

These experiments were done to learn whether Mycoplasma pulmonis infections of the respiratory tract of rats can potentiate "neurogenic inflammation" and whether this potentiation is amplified by factors that exacerbate the infections. Pathogen-free F344 rats were inoculated intranasally with M. pulmonis or with sterile culture medium and then lived for 4 wk in an ammonia-free atmosphere or in air containing ammonia (100 parts per million). Neurogenic inflammation was evoked by an intravenous injection of capsaicin, and 5 min later the magnitude of the response was quantified by measuring the amount of extravasation of two tracers, Monastral blue pigment and Evans blue dye. We found that vascular permeability in the tracheas of all rats was normal in the absence of capsaicin. However, a 75-micrograms/kg dose of capsaicin, which caused almost no extravasation of Evans blue in the tracheas of pathogen-free controls (17 +/- 3 ng/mg; mean +/- SE), produced extensive extravasation in the infected rats (135 +/- 18 ng/mg; P less than 0.001). Similarly, this dose of capsaicin produced 30 times as much Monastral blue extravasation in the infected rats (area density = 47 +/- 8% of surface area) as it did in the pathogen-free rats (1.6 +/- 0.5%; P less than 0.001), a difference that resulted from increases in the number of Monastral blue-labeled postcapillary venules and in the amount of labeling per venule. Exposure of the infected rats to ammonia exacerbated the infections, further increased the number of Monastral blue-labeled vessels and the amount of labeling per vessel, and made the rats so sensitive to capsaicin that a normally tolerable dose of 150 micrograms/kg i.v. caused fatal apnea. Ammonia did not have these effects in pathogen-free rats. We conclude that M. pulmonis infections of the airway mucosa cause a potent, long-lasting potentiation of neurogenic inflammation, which results in part from an increase in the number and responsiveness of mediator-sensitive postcapillary venules. These changes can be amplified by environmental factors such as ammonia which exacerbate the infections.

Mechanism of Tachyphylaxis Associated with Neurogenic Plasma Extravasation in the Rat Trachea

Electrical stimulation of the vagus nerve of rats is known to produce plasma extravasation in the trachea, presumably by releasing substance P or other tachykinins from sensory nerves. We sought to determine whether the tachyphylaxis that develops after prolonged vagal stimulation results from an inability of sensory nerves to release tachykinins or from an inability of tracheal blood vessels to respond to tachykinins. To induce tachyphylaxis, we electrically stimulated the right vagus nerve of Long-Evans rats for 5 min (5 V, 1 ms, 20 Hz). Then, 10 min later, we gave intravenous injections of capsaicin (0.3 mumol/kg), histamine (18 mumols/kg), or substance P (2.2 nmol/kg), which produce equivalent amounts of plasma extravasation as assessed by the extravasation of Evans blue dye. We found that vagal stimulation reduced the amount of dye extravasation produced by capsaicin but not the amount produced by either histamine or substance P. We also found that pretreating neonatal rats with capsaicin, which destroys tachykinin-containing sensory nerves, reduced the amount of dye extravasation produced by capsaicin but not the amount produced by either histamine or substance P. This finding suggests that capsaicin produces plasma extravasation in the trachea by releasing tachykinins from sensory nerves, whereas histamine and substance P do so by acting directly on tracheal blood vessels. Taken together, our results indicate that prolonged vagal stimulation reduces the ability of sensory nerves to release tachykinins but that tracheal blood vessels remain fully responsive to both histamine and substance P.

Reflex responses on blood pressure and renal nerve activity to local intra-arterial injection of capsaicin in anesthetized dogs.

The present study was performed to examine effects of intra-arterial and intravenous injection of capsaicin on efferent renal sympathetic nerve activity (RNA) in pentobarbital anesthetized dogs. In animals with intact baroreceptors, intra-arterial injection of capsaicin (4 +/- 1 microgram) produced significant increases in mean blood pressure (MBP) and heart rate (HR) by +43 +/- 8 mmHg and +33 +/- 15 beats/min at 20 s after the injection, respectively. There followed a recovery toward the control so that 60 s after the intra-arterial injection changes in MBP and HR were +8 +/- 5 mmHg and +18 +/- 8 beats/min, respectively. RNA showed a biphasic response, comprising of an initial increase (+52 +/- 28% of the control at 20 s) followed by a decrease by -30 +/- 8% of the control 60 s after the injection. Following bilateral cervical vagotomy the initial increase in RNA did not differ significantly from the value of RNA in animals with intact baroreceptors. However, the later decrease in RNA reversed above the control so that 60 s after the injection change in RNA was +5 +/- 10% of the control. Complete baroreceptor denervation showed a similar response on RNA in the vagotomized animals (+49 +/- 9 and +14 +/- 10% of the control at 20 and 60 s after the injection, respectively). In contrast, intravenous injection of capsaicin (6 +/- 1 microgram/kg) resulted in significant decreases in MBP (-19 +/- 11 mmHg) and HR (-9 +/- 4 beats/min). RNA at 20 s after the injection showed a unidirectional decrease by -16 +/- 7% of the control in animals with intact baroreceptors. These responses reversed above the control after cervical vagotomy. Thus, these data indicate that activation of C-fiber afferents in skeletal muscles by intra-arterial injection of capsaicin results in renal sympathoexcitation, and that the later sympathoinhibition is mediated by combined activation of systemic baroreceptors.

Inhibition of neutral endopeptidase potentiates neurogenic inflammation in the rat trachea.

The present study was performed to determine whether neurogenic inflammation in the rat trachea can be exaggerated by inhibiting neutral endopeptidase, an enzyme that degrades tachykinins that are believed to mediate neurogenic inflammation. Neurogenic inflammation was produced by antidromic electrical stimulation of one vagus nerve (2.5 Hz, 1 ms, 5 V for 5 min) in the presence of atropine or by an intravenous injection of capsaicin (100 micrograms/kg). Neutrophils that adhered to the endothelium of venules were visualized and counted in tracheal whole mounts that were stained by a histochemical reaction for myeloperoxidase. Neural inflammation increased the number of adherent neutrophils. Pretreatment with the neutral endopeptidase inhibitor phosphoramidon (1.0 or 2.5 mg/kg iv) increased neutrophil adhesion induced by neural inflammation. As assessed by the amount of extravasation of Monastral blue pigment, neural inflammation also increased vascular permeability, and this change was potentiated by phosphoramidon. These results are consistent with the concept that neuropeptides released from sensory nerves in the tracheal mucosa cause neutrophils to adhere to venules and increase vascular permeability and that these effects are modulated by neutral endopeptidase.

Lung C-fibre receptor activation and defensive reflexes in anaesthetized cats.

1. With pentobarbitone-anaesthetized cats we have elicited cough reflexes from the tracheobronchial tree and the larynx, and the aspiration and sneeze reflexes from the nasopharynx and the nose respectively. The reflexes were induced by mechanical stimulation of the mucosa, before and during activation of pulmonary C-fibre receptors by intravenous injections of capsaicin or phenylbiguanide. 2. During the 20-30 s apnoea due to C-fibre stimulation, the cough reflex from both sites and the sneeze reflex were completely abolished, whereas the aspiration reflex response was approximately halved. Reflex contractions of genioglossus muscle still occurred at this time, but were far weaker than in the control state. 3. During the rapid shallow breathing that immediately followed apnoea due to C-fibre receptor stimulation, the defensive reflexes recovered: the aspiration and sneeze reflexes fully and the cough reflexes to about half of the control response. 4. Acute hypotension due to haemorrhage, of a size considerably greater than that due to stimulation of the pulmonary C-fibre receptors, caused no significant inhibition of the cough reflex from the tracheobronchial tree. 5. We conclude that the pulmonary C-fibre reflex powerfully inhibits airway defensive reflexes, and that its activation is unlikely to contribute positively to coughing induced by aerosols of capsaicin and similar agents.

Pulmonary depressor reflex elicited by capsaicin in conscious intact and lung-denervated dogs.

A pulmonary depressor reflex has been shown to be elicited in anesthetized cats, dogs, and rats by intravenous injection of capsaicin. The effects observed include apnea, hypotension, and bradycardia with some investigators reporting tachypnea following the apneic period. We investigated the response to a bolus injection of capsaicin (20 micrograms/kg) into the cephalic vein in 11 conscious beagle dogs. Five control dogs underwent sham thoracotomies, and six dogs underwent selective denervation of the lungs. A low dead-space latex rubber mask was used to monitor ventilation, and arterial blood pressure was obtained by catheterizing an exteriorized carotid artery. In four of the five control dogs the observed response was apnea concomitant with hypotension and bradycardia, followed by tachypnea. In five of the six lung-denervated dogs there was a slight tachypnea along with hypertension. It is concluded that the pulmonary depressor reflex can be elicited in conscious dogs by intravenous injection of capsaicin but is absent in lung-denervated dogs.

Cardiovascular reflexes mediated by capsaicin sensitive cardiac afferent neurones in the dog

Reflex circulatory changes mediated by capsaicin sensitive cardiac afferent neurones were studied in anaesthetised, open chest dogs. Application of capsaicin to the epicardium of the left ventricle, either in single doses (0.01-100 micrograms) or by superfusion (20 micrograms X min-1), consistently resulted in dose related increases in blood pressure and heart rate. These responses were not affected by bilateral vagotomy but were abolished or reversed by bilateral sectioning of the upper thoracic (T1-T4) white rami communicantes and stellectomy. Injection of capsaicin (0.3-1 microgram X kg-1) into the left circumflex coronary artery caused either systemic hypotension and bradycardia (70.3% of experiments), a pressor response associated with tachycardia (13.5%), or a biphasic effect with an initial rise and then fall in blood pressure and heart rate (16.2%). With intravenous injections of capsaicin (3-5 micrograms X kg-1) the response was invariably cardioinhibitory and depressor. The reflex bradycardia and hypotension evoked with either intracoronary or intravenous injections of capsaicin were reversed after bilateral vagotomy to increases in cardiac rate and blood pressure. The post-vagotomy tachycardia occurring with intracoronary capsaicin could be abolished by beta adrenoceptor blockade with propranolol (0.5 mg X kg-1 iv), whereas ganglionic transmission blockade with pentolinium (0.5 mg X kg-1 iv) eliminated both the tachycardia and pressor effects. The results indicate that in the dog's heart capsaicin sensitive afferent neurones capable of affecting the circulatory system have both vagal and spinal sympathetic origin. It is suggested that capsaicin induced excitatory cardiogenic reflex is nociceptive in nature and may involve activation of substance P containing afferent fibres incorporated in cardiac sympathetic nerves.