Substance P Antagonist Research Articles

Corticotropin releasing factor and substance P mediate the nucleus amygdaloideus centralis-nucleus ventromedialis-nucleus dorsomedialis pressor system

Prolonged emotional stress is an important factor in the development of neurogenic hypertension, but its mechanism is still unclear. The purpose of the present study is to analyze the possible neural basis of hypertension induced by prolonged emotional stress. In the brain many nuclei are involved in emotional reaction, stress or defense response; among them the nucleus amygdaloideus centralis (AC) is the most important one which widely connects with other nuclei controlling emotion and stress, such as nucleus ventromedialis (NVM), nucleus dorsomedialis (NDM), nucleus paraventricularis (NPV) etc. These nuclei contain corticotropin releasing factor (CRF)- and substance P (SP)-immunoreactive cell bodies, nerve terminals and corresponding receptors. Our previous and present studies showed that microinjection of CRF or SP into these nuclei induced pressor responses. These data imply that excitation of the AC can activate many nuclei controlling emotion and stress via CRF and SP, and excessive activities of these nuclei may be the neural basis of hypertension induced by prolonged emotional stress. The present study revealed that (1) the AC pressor response to glutamate (Glu) could be reduced by preinjection of CRF antagonist (α-Helical CRF[9-41] or SP antagonist ([ d-Pro 2, d-Phe 7, d-Trp 9]-substance P) into bilateral NVM, (2) the NVM pressor response to Glu were decreased by pretreatment of the NDM with CRF- or SP-antagonist, (3) the AC-, NVM- or NDM-pressor responses were all attenuated by preinjection of CRF- or SP-antagonist into bilateral NPV or rostral ventrolateral medulla (RVL). The results indicate that excitation of the AC can indirectly activate the NPV and RVL to evoke pressor response via the NVM–NDM, CRF and SP are transmitters in each connection of this pathway; this is one component of the mechanism underlying the AC pressor response. Taken together with the findings of our previous studies, it provides neurophysiological basis for the above-mentioned implications.

Modulation of sympathetic nerve activity by perivascular sensory nerves in the arterioles of the guinea-pig small intestine

Objective: The aim of this study was to assess the role of perivascular sensory nerves in modulating constrictions of intestinal submucosal arterioles. Methods: Arteriole constrictions were induced either by nerve released adenosine 5′-triphosphate (ATP) or exogenous ATP or phenylephrine (PE). Individual nerve shocks were used to elicit excitatory junction potentials (EJPs) in the arteriole smooth muscle whereas trains of stimuli were used to evoke transient constrictions of the arteriole. Effects of the sensory neurotoxin, capsaicin, were examined on constrictions and EJPs. Results: Pre-treatment of the arteriole preparation with capsaicin did not cause any significant change in the amplitude of arteriole constrictions to exogenously applied ATP or PE. However, there was a significant increase in the amplitude of neurally evoked arteriole constrictions and EJPs, without a significant change in the decay time constant ( τ decay) of the EJPs. Exogenous application of calcitonin gene related peptide (CGRP) significantly decreased τ decay of the EJPs without affecting their amplitude, whereas substance P (SP) significantly decreased the amplitude of EJPs without affecting τ decay. Both, CGRP and SP, decreased the amplitude of neurally evoked and ATP induced constrictions. Whilst the inhibitory effects of CGRP on evoked and ATP induced constrictions were not significantly different, the reduction in evoked constrictions obtained with SP was significantly greater than the reduction in ATP induced constrictions. SP antagonist significantly increased the amplitude of neurally evoked constrictions. Conclusions: It is concluded that capsaicin-sensitive afferents inhibit the release of transmitter from perivascular sympathetic nerves via the prejunctional modulatory action of SP. The other putative sensory neurotransmitter, CGRP, appears to act postjunctionally on the arteriole smooth muscle.

Acute management of migraine: triptans and beyond.

Migraine is a paroxysmal disorder characterized by attacks of headache, nausea, vomiting, photophobia and phonophobia, and malaise. This review summarizes new treatment options for the therapy of acute attacks. Sumatriptan was the first specific serotonin-1B/D agonist for the treatment of acute migraine attacks. Apart from the oral and subcutaneous formulation, it is also available as nasal spray and suppository. The other new migraine drugs zolmitriptan, naratriptan, rizatriptan and eletriptan differ in their pharmacological profiles, which translates into minor differences in efficacy, headache recurrence and side-effects. Importantly, in clinical practice individual patients may show a preference for one treatment over another. New drugs in migraine treatment include substance-P antagonists, nitric oxide synthetase inhibitors and calcitonin gene-related peptide antagonists.

Functional neurokinin NK-1 receptor expression in rat peritoneal mast cells.

Recently, Ogawa et al. [17] reported that the peritoneal mast cells (PMCs) of rats can release histamine by substance P (SP) in a receptor-dependent manner. In the present study, we confirmed and extended their findings. PMCs were isolated from six strains of rats. In some experiments, peritoneal cells in the non-MC fraction were used. PMCs were incubated with SP, neurokinin (NK) receptor agonists or antagonists, and histamine content in the supernatant was measured. In the binding assay, PMCs were incubated with [125I]BH-SP together with SP or NK receptor antagonists. NK-1 receptor mRNA was detected using a reverse transcription-polymerase chain reaction (RT-PCR) assay. PMCs from Slc: Wistar and F344/NSlc were highly sensitive to SP, leading to histamine release, whereas those from Slc:SD and three other strains were not. PMCs from Slc:Wistar and F344/NSlc also released histamine in the presence of an NK-1 agonist. The histamine release induced by SP and the NK-1 agonist was inhibited by the NK-1 receptor antagonists, FK888 and CP-99,994. [125I]BH-SP binding experiments revealed that PMCs from Slc:Wistar rats possessed a single high affinity binding site for SP and that the binding was blocked by NK-1 receptor antagonists. Peritoneal cells in the non-MC fraction exhibited no appreciable binding. In the RT-PCR assay, expression of NK-1 receptor mRNA was evident in Slc:Wistar PMCs, but not in the non-MC fraction from Slc:Wistar or Slc: SD PMCs. These data demonstrate the existence of functional NK-1 receptors on freshly isolated PMCs in at least some strains of rats.

A Substance p Agonist Acts as an Adjuvant to Promote Hapten-Specific Skin Immunity

Because substance p (SP) has been reported to be released from cutaneous sensory nerve endings after hapten application, we determined whether SP participates in contact hypersensitivity (CH) induction by using a SP agonist, GR73632 or delta-Aminovaleryl [Pro9, N-Me-Leu10]-substance P(7-11) and a SP antagonist, spantide I. When injected intradermally, SP agonist enhanced CH induced by conventional, but not optimal, sensitizing doses of hapten. By contrast, SP antagonist inhibited the induction of CH by optimal sensitizing doses of hapten. Moreover, SP agonist promoted CH induction and prevented tolerance when hapten was painted on skin exposed to acute, low-dose ultraviolet-B radiation. Intradermally injected SP agonist altered neither the density nor the morphology of epidermal Langerhans cells, implying that SP agonist enhanced the generation of hapten-specific immunogenic signals from the dermis. It is proposed that SP is a natural "adjuvant" that promotes the induction of CH within normal skin. Although exogenous SP agonist can prevent impaired CH and tolerance after ultraviolet-B radiation, the susceptibility of native SP to local neuropeptidases renders the neuropeptide unable to prevent the deleterious effects of ultraviolet-B radiation on cutaneous immunity.

Effect of intravenous substance P on laryngeal adductor activity in young dogs.

Reflex laryngeal adduction is a component of both the laryngeal chemoreflex and the esophagolaryngeal adductor reflex, two life-threatening reflexes that occur in immature animals. These two reflex responses are also thought to exist in infants and may play a role in causing life-threatening laryngospastic events and perhaps sudden infant death syndrome. Identifying neurotransmitters that mediate laryngeal adduction is important to understanding the mechanism of reflex laryngeal responses and to identifying potential means of pharmacologic prevention. Substance P (SP), a tachykinin, putatively functions as a sensory neurotransmitter and may play a role in mediating laryngeal reflexes. Substance P-immunoreactive-like fibers and receptors are present in the subepithelial tissues of the larynx, the vagus nerves, the nodose and jugular ganglia, and the vagal brain stem nuclei. In this investigation, the effect of SP infusion on laryngeal motor activity in an in vivo model is reported. Substance P was infused intravenously into 8 puppies (20 to 133 days of age, mean 81.2), on a mean of 3.0 occasions (range 1 to 6). Cardiovascular, respiratory, arterial blood gas, and cricothyroid (CT), thyroarytenoid (TA), and genioglossus electromyographic (EMG) responses to infusion of the tachykinin were recorded and subsequently analyzed. The SP infusion induced a marked increase in CT or TA EMG activity in 23 of 24 studies, and the increase was typically apparent within 60 seconds of the infusion. An increase in genioglossus EMG activity did not occur. An immediate, profound decrease in mean arterial pressure and an increase in respiratory rate and depth of chest wall excursion accompanied the laryngeal response. Arterial blood gas values remained unchanged (p > .05). The laryngeal adductor response to SP infusion was blocked when animals were pretreated with a systemic SP antagonist (Pfizer CP-96,345). This study demonstrates that peripheral infusion of the tachykinin SP induces a marked increase in laryngeal adductor activity. The response may be blocked by pretreatment of animals with a systemic SP antagonist. Because SP is thought to act primarily as a sensory neurotransmitter, these findings may be important in understanding the mechanism of reflex laryngeal adductor responses.

SR140333, a substance P receptor antagonist, influences morphological and motor changes in rat experimental colitis.

The etiology of inflammation, edema, and smooth muscle contraction characteristic of inflammatory bowel disease is not clearly understood. There is evidence that several neuropeptides, including substance P (SP), may play a role. In this study we evaluated the ability of a SP-antagonist (SR140333) to modify the course of experimental colitis induced in the rat by trinitrobenzene sulfonic acid (TNB). Colitis was induced in 24 rats using TNB applied by intrarectal enema. Twelve TNB-treated rats received SR140333, 0.1 mg/kg intraperitoneally, 30 min before the administration of TNB and every 48 hr until death. Twelve rats receiving only intrarectal 0.9% saline served as controls. Rats of each group were killed after 14 days. At day 14, the control group showed no signs of inflammation whereas the TNB-treated rats without SR140333 treatment exhibited a well-established colitis. The TNB-treated group had a higher level of inflammation, as seen histologically and by the significantly greater weight of colon strips, compared to the controls (0.30 +/- 0.09 g vs 0.13 +/- 0.03 g, P < 0.001) and to the SR140333-treated rats (0.30 +/- 0.09 g vs 0.14 +/- 0.05 g, P < 0.001). In addition, smooth muscle contractility was significantly reduced in the inflamed colons of TNB-treated rats when compared with the controls (carbachol: 42.7 +/- 20.3 vs 254.2 +/- 69.78 mg/mm2; SP: 18.5 +/- 10.02 vs 89.45 +/- 23.17 mg/mm2; KCl: 11.4 +/- 2.2 vs 98.32 +/- 33.57 mg/mm2, P < 0.01). However, SR140333-treated rats showed a recovery from inflammation and motor alterations caused by TNB (carbachol: 150.9 +/- 46.1 mg/mm2, P < 0.01; SP: 32.5 +/- 9.4 mg/mm2, P < 0.05; KCl: 125.7 +/- 36.1 mg/mm2, P < 0.01). In conclusion, treatment with SP antagonist SR140333 reduces the severity of colitis and has beneficial effects on the concomitant alterations of contractility. Thus, the blockade of substance P may represent a possibility in the treatment of intestinal inflammation.

Substance-P antagonists: a new treatment for depression?

Since its discovery by von Euler 60 years ago, substance P has exercised the minds of pharmacologists and become of great interest for drug discovery. Because the peptide is a transmitter in pain endings in the spinal cord, pharmaceutical companies searching for novel analgesics have tried to synthesise specific and selective antagonists for the substance-P receptor, also known as the neurokinin 1 (NK1) receptor. Since the first report on such a compound in 19911 several substance-P antagonists have been synthesised, but none has been effective in reducing pain.

Derivation of a three-dimensional pharmacophore model of substance P antagonists bound to the neurokinin-1 receptor.

Constrained systematic search was used in an exhaustive conformational analysis of a structurally diverse set of substance P (SP) antagonists to identify a unique hypothesis for their bound conformation at the neurokinin-1 receptor. In this conformation, two aromatic groups essential for high affinity adopt a perpendicular or edge-on arrangement. This pharmacophore hypothesis for the receptor-bound conformation was used in a comparative molecular field analysis (CoMFA) of an expanded set of SP antagonists, and the predictive ability of the resulting three-dimensional quantitative structure-activity relationship (3D-QSAR) was evaluated against a test set of SP antagonists different from those in the training set. This CoMFA model based on the Constrained Search alignment yielded significant cross-validated, conventional, and predictive r2 values equal to 0.70, 0.93, and 0.82, respectively. For comparison, the SP antagonists were forced into an alternative poorer alignment in which the two aromatic rings were parallel and then subjected to a CoMFA analysis. Both the parallel and perpendicular arrangements of the aromatic rings are seen in X-ray structures of SP antagonists and have been proposed as candidates for the receptor-bound conformation. The parallel (or stacked) conformation yielded a poorer correlation with a cross-validated r2 = 0.57, a conventional r2 = 0.90, and a predictive r2 = 0.78. Our results indicate that although both alignments could generate a reasonable CoMFA correlation, the stacked conformation is unlikely to be the receptor-bound conformation, as the covalent structure of the antagonists precludes a common geometry in which the aromatic rings are stacked.

NK1 receptor antagonist blocks angiotensin II responses in renin transgenic rat medulla oblongata.

Angiotensin (Ang) II increases substance P (SP) efflux from perfused slices of medulla oblongata, and a peptide antagonist of SP, [Leu11,psiCH2NH10-11]SP, blocks the acute hypotension and bradycardia caused by Ang II injected into the nucleus tractus solitarii (nTS) of Harlan Sprague-Dawley (SD) rats. We investigated whether the same relationships exist in (mRen2)27 renin transgenic (TG) rats, which have chronic elevations of medullary tissue Ang II levels. Ang II increased SP efflux (48% above control; P<0.01) from slices of medulla prepared from 8- to 12-week old male TG rats. Injections of Ang II (250 fmol in 30 nL) into the nTS of chloralose-urethane anesthetized TG rats produced a significant increase in pressure of 7+/-2 mm Hg before a 13+/-3 mm Hg fall in pressure. Ang II induced similar depressor responses in Hannover SD rats but no increase in pressure. After nTS injection of the NK1-selective SP antagonist CP-96,345 (30 pmol in 60 nL), Ang II-induced hypotension was blocked in both groups, as was the pressor component in hypertensive rats. Hypotensive and bradycardic effects of glutamate (0.6 nmol in 30 nL) injected into the nTS were not altered by CP-96,345. In vitro receptor autoradiography showed that the SP antagonist (10 or 100 microM) did not compete for 125I-Ang II binding in the dorsal medulla, a result suggesting that it did not interact directly with Ang II receptors. Thus, the nTS cardiovascular effects of Ang II are mediated by SP in both normotensive rats and a model of hypertension with altered endogenous levels of Ang II. These findings link Ang II-induced effects on SP release from brain slices of the medulla oblongata to acute cardiovascular actions of the peptide through an NK1 receptor.

An antagonist of substance P N-terminal fragments, d-substance P(1–7), reveals that both nociceptive and antinociceptive effects are induced by substance P N-terminal activity during noxious chemical stimulation

Substance P (SP) N-terminal fragments are known to alter nociception when injected intrathecally or when released in response to capsaicin. However, it is not known whether a sufficient concentration of SP N-terminal metabolites accumulate during noxious stimulation to modulate nociception. To test this, we examined the effect of the SP(1–7) antagonist, d-SP(1–7), injected intrathecally in mice, on two nociceptive assays that are differentially affected by exogenous SP(1–7): acetic acid-induced writhing that is inhibited and formalin-induced behaviors that are enhanced by SP(1–7). One nmol of d-SP(1–7) is sufficient to block the acute (30 min) antinociceptive effects of SP(1–7) on writhing. When injected alone at much higher doses (10–100 nmol), d-SP(1–7) inhibited writhing. In the formalin assay, SP(1–7) had no acute effect (30 min) on responses during Phase 1 at any dose tested, but d-SP(1–7) increased responses 5 min after injection of low (2–1000 pmol), but not high doses (10 and 100 nmol). Twenty-four hours after injection of SP(1–7), writhing was inhibited and formalin responses were increased. d-SP(1–7) prevented these effects of SP(1–7) but had no effect when injected alone, indicating that there is no tonic SP N-terminal activity in mice not exposed to noxious stimuli. Thus, acetic acid and formalin each induce endogenous SP N-terminal activity, respectively, producing a pro-nociceptive effect that is relatively insensitive to d-SP(1–7) and antinociception that is very sensitive to inhibition by d-SP(1–7).

The changes in pulmonary C-fiber activity and lung mechanics induced by vagal stimulation in rabbits

The effect of unilateral stimulation of the vagus nerve on pulmonary C-fiber activity, total lung resistance (R L) and dynamic lung compliance (Cdyn) was studied in anesthetized, artificially ventilated rabbits. Vagal stimulation (4–30 Hz) increased both pulmonary C-fiber activity and R l but decreased Cdyn, and these changes were frequency-dependent. Atropine (2 mg kg ) blocked the responses of pulmonary C-fiber, R L and Cdyn to vagal stimulation at 4–30 Hz. Stimulation of pulmonary C-fiber activity by the higher volume-induced lung inflation was not significantly altered by atropine (2 mg kg ). A selective SP antagonist [D-Pro 2, D-Try 7,9]-SP (0.5 mg kg ) inhibited bronchoconstrictor responses to vagal stimulation at 30Hz but had no significant effect on those to the stimulation at 4–17 Hz. These results suggest that excitation of the rabbit pulmonary C-fiber activity by unilateral vagus nerve stimulation would occur as a result of the mechanical stimuli via a cholinergic neurotransmission and that during the stimulation at 30 Hz this excitation partly involves the local release of substance P (SP) to promote acetylcholine outflow.

Synergistic Effect of Substance P with Epidermal Growth Factor on Epithelial Migration in Rabbit Cornea

In order to investigate the role of neural regulation in corneal epithelial healing, we examined the effect of substance P (SP) on corneal epithelial migration using an organ culture system of rabbit corneas. We investigated the synergistic effects of SP with (1) growth factors: epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), and transforming growth factor-β(TGF-β); (2) extracellular matrix proteins: fibronectin, vitronectin, laminin, and collagen type IV; and (3) cytokines: interleukin-1α (IL-1α), IL-1β, and interleukin-6 (IL-6). Rabbit corneal blocks were cultured in the absence or presence of various reagents for 24 hr. The corneal blocks were then fixed, dehydrated, embedded in paraffin and stained by hematoxylin-eosin, and the length of the path of epithelial migration was measured. The addition of SP alone, at concentrations up to 50 μg ml−1, did not affect epithelial migration. EGF, fibronectin, vitronectin, collagen type IV, and IL-6 stimulated epithelial migration, but bFGF, TGF-β, laminin, IL-1α, and IL-1βdid not. The stimulatory effect of EGF on the epithelial migration was enhanced by the presence of SP. This synergistic effect of SP and EGF on corneal epithelial migration was abolished by the addition of an SP antagonist or enkephalinase. Other neurotransmitters (vasoactive intestinal peptide, calcitonin gene-related peptide, acetylcholine chloride, norepinephrine, serotonin) and tachykinins (neurokinin A, neurokinin B, kassinin, eledoisin, physalaemin) were examined, but none exhibited a synergistic effect with EGF. Interestingly, EGF alone stimulated the incorporation of3H-thymidine into corneal epithelial cells, but the addition of SP with EGF did not enhance this effect. These results demonstrate that SP enhanced the EGF stimulation of corneal epithelial migration in vitro in a specific manner, suggesting a possible role of SP as a modulator of epithelial wound healing.

Tandem Michael addition and azanorbornane substance-P antagonists

Tandem Michael addition product 5 provides a template for azanorbornane derivatives that display potent substance P receptor affinity. The advantages of this synthesis include: the straightforward preparation of both syn and anti azanorbornane diastereomers, access to two, and perhaps more, substituents at C-2 ( a and b series), and the straightforward functionalization of the amine at C-3. Mechanistic studies suggest that the second 1,4-addition proceeds kinetically via syn-addition of the nitronate and ester. Conflicting results in the literature stem from a previously unobserved thermodynamic equilibrium which is described herein.

Synergistic Interactions between Neuropeptide and Histamine on the Capillary Permeability in Rat Skin: Evaluation by Reflectance Spectrophotometry

Effects of neuropeptides on capillary permeability and their interactions with histamine (HIS) in rat skin were investigated. The capillary permeability was measured continuously by reflectance spectrophotometry after intravenous (iv) injection of Evans blue dye. The capillary permeability was increased dose-dependently by the intradermal injection of HIS (0.3–100 μg/site) and substance P (SP; 25–250 ng/site). Calcitonin gene-related peptide (80–800 ng/site) elicited a significant but less increase than did SP. Capsaicin (30 μg/site) also increased capillary permeability slightly but significantly, suggesting the release of endogenous neuropeptides. Both diphenhydramine (DPH; 3 mg/kg, iv) and cimetidine (CIM; 30 mg/kg, iv) reduced HIS-induced responses. DPH also reduced the SP-induced response significantly, but CIM did not. An SP antagonist, [d-Pro2,d-Trp7,9]-SP (1.3 mg/kg, iv), reduced not only SP- but also HIS-induced responses. Furthermore, the HIS-induced response was attenuated by pretreatment with epicutaneous capsaicin for 4 days, depleting endogenous SP. These results delineate the synergistic interactions between SP and HIS in rat skin and suggest the participation of neuropeptides in increasing capillary permeability.

Increased substance P responses in dorsal root ganglia and intestinal macrophages during Clostridium difficile toxin A enteritis in rats.

Previously we reported that pretreatment of rats with the substance P (SP) antagonist CP-96,345 inhibits the enterotoxic responses following administration of toxin A from Clostridium difficile into ileal loops, indicating that SP participates in the intestinal responses to this toxin. We now report that injection of toxin A into rat ileum causes a rapid increase in SP content in lumbar dorsal root ganglia (DRG) and mucosal scrapings 30-60 min after toxin A administration. Toxin A-mediated fluid secretion, mannitol permeability, and ileal histologic damage is significantly increased only after 2 hr. Toxin A also causes an increase in the abundance of SP mRNA in lumbar DRG and ileal mucosa as measured by reverse transcription-PCR. Lamina propria macrophages (LPMs) obtained from toxin A-injected loops release greater amounts of tumor necrosis factor alpha (TNFalpha) and SP as compared with LPMs isolated from buffer-injected loops (P < 0.01). Pretreatment of rats with the SP antagonist CP-96,345 inhibits toxin A-mediated TNFalpha release from isolated LPMs, whereas an inactive enantiomer (CP-96,344) of the SP antagonist has no effect. LPMs obtained from toxin A-injected ileal loops incubated in vitro with SP (10(-8) to 10(-9) M) show enhanced TNFalpha secretion, whereas LPMs isolated from buffer-injected loops do not respond to SP. In addition, LPMs obtained from toxin A-injected ileal loops incubated in vitro with CP-96,345 showed a diminished TNFalpha release. Our results indicate that activated LPMs secrete SP during toxin A enteritis that can lead to secretion of cytokines, suggesting an autocrine/paracrine regulation of cytokine secretion by SP from LPMs during intestinal inflammation.

A substance P antagonist, RP-67,580, ameliorates a mouse meningoencephalitic response to Trypanosoma brucei brucei.

Mice infected with the protozoan parasite Trypanosoma brucei brucei and treated subcuratively with the trypanocidal drug diminazene aceturate develop an acute inflammatory meningoencephalitis with associated astrocytic proliferation. This reaction is very similar to that seen in the fatal posttreatment reactive encephalopathies that can occur in human African trypanosomiasis. The 11-amino acid neuropeptide substance P (SP) has recently been identified as a mediator in many inflammatory responses, and the development of potent, highly specific, nonpeptide SP antagonists has provided a new opportunity to investigate the possible involvement of SP in a variety of pathological conditions. We therefore postulated that SP may play a role in the development of the posttreatment inflammatory encephalopathy found in this experimental mouse model of African trypanosomiasis. In the present study RP-67,580, a SP antagonist that binds specifically to NK-1 receptors, was given intraperitoneally at a dose of 2 mg/kg twice daily to mice in which a severe meningoencephalitis had been produced. A significant reduction in both the severity of the inflammatory response (P = 0.0001) as well as the degree of astrocyte activation (P < 0.001) was found in the brains of these animals as compared with control mice that had not received RP-67,580. An inactive enantiomer of this SP antagonist, RP-68,651, had no effect on the central nervous system inflammatory reaction. We conclude from these findings that the neuropeptide SP plays a key role in the development of the severe central nervous system inflammatory response associated with African trypanosomiasis.

Response of isolated human neurogenic bladders to tachykinins.

Tachykinins act as stimulants of the isolated urinary bladder. However, the efferent role of tachykinins on detrusor function is controversial. We investigated the contractility of isolated human neurogenic bladders taken from patients with myelomeningocele or sacral agenesis, and tested the response to neurokinin A (NKA), neuromedin K (NKB), and substance P (SP). The contractile strengths were compared with normal controls. All tachykinins investigated induced significant contractions in both neurogenic and control bladders. The rank order of contractile potency was the same in both groups, namely, NKA > NKB > SP. The contraction induced by SP was not affected by atropine, but was completely blocked by [Sar9,Met(O2)11]-SP (a SP antagonist). Responses to electrical field stimulation were not changed by the SP antagonist. The contractile magnitude to field stimulation was also not altered by administration of 10(-6) M tachykinins. Responses of the neurogenic bladder to NKA and SP were significantly greater than the control. There were no differences in the response to KCl administration between the 2 groups. We conclude that hypersensitivity to NKA and SP in neurogenic bladders may contribute to bladder dysfunction in patients with sacral cord lesions.

Substance P augments interleukin-10 and tumor necrosis factor-α release by human cord blood monocytes and macrophages

We have investigated the effects of SP on the constitutive and/or lipopolysaccharide (LPS)-induced expression of interleukin-10 (IL-10) and tumor necrosis factor (TNF-α) in both freshly isolated cord blood monocytes (FICBM) and cord blood monocyte-derived macrophages (CBMDM). The cells were treated with SP at various concentrations (10 −14 to 10 −6 M) in the presence or absence of LPS and culture supernatants were analyzed for IL-10 and TNF-α as measured by an enzyme immunosorbent assay (ELISA). FICBM and CBMDM treated with SP alone increased TNF-α secretion. The stimulatory effects of SP on TNF-α secretion are inhibited by a anti-SP polyclonal antibody and SP antagonists, spantide ([D-Arg-1-D-Trp-7-D-Trp-9-Leu-11]-SP) and CP-96,345 (a nonpeptide antagonist of the SP receptor). Although the treatment with SP alone did not enhance IL-10 secretion by both freshly isolated and cultured cord monocytes, treatment with SP in combination with LPS leads to a synergistic interaction in upregulation of IL-10 secretion. Fragments of SP (SP 1–4 and SP 5–11) in the presence or absence of LPS show little effects on IL-10 secretion by FICBM. SP reverses the inhibitory effect of IFN-γ on LPS-induced IL-10 secretion by FICBM. In addition, the two SP antagonists and the anti-SP polyclonal antibody blocked the SP effect on IL-10 secretion by FICBM, indicating that these effects are specific and SP receptor mediated. Thus, SP is likely to play an important role in certain inflammatory conditions in the immune and nervous systems.

Nitric oxide mediates long-term hyperalgesic and antinociceptive effects of the N-terminus of substance P in the formalin assay in mice

Conditions such as hyperalgesia can occur days or months after the noxious insult. Substance P (SP) is released in response to noxious stimuli. Given the long-term effects of the N-terminus of SP on putative nociceptive transmitters, we investigated changes in formalin-induced nociception following an accumulation of SP N-terminal metabolites in mice. Pre-treatment with the N-terminal metabolite of SP, SP(1–7), was without effect when injected intrahecally (i.t.) 5 or 30 min before formalin. However, at 24 h, SP(1–7) increased behaviors during Phase 1, indicating hyperalgesia, and attenuated Phase 2 responses, consistent with antinociception. The nitric oxide (NO) synthase inhibitor, Nω-nitro- l-arginine methyl ester HCl ( l-NAME), blocked both hyperalgesic and antinociceptive effects when co-injected with SP(1–7). Consistent with a NO-mediated pathway, l-arginine ( l-arg), the N-terminal amino acid of SP and precursor to NO, mimicked the antinociceptive effect of SP(1–7) on Phase 2. The hyperalgesic effect of SP(1–7) in Phase 1, which was not mimicked by l-arg, was prevented by d-SP(1–7), a SP(1–7) antagonist. Thus, SP(1–7) modulates nociception via two distinct NO-mediated pathways. When injected for 7 days, tolerance developed to the antinociceptive effect of SP(1–7) on Phase 2, but not to the hyperalgesic effect on Phase 1. Intraperitoneally injected SP(1–7) also produced hyperalgesia during Phase 1, to which tolerance developed following seven daily injections. Together, these data support the hypothesis that an accumulation of SP N-terminal metabolites, either peripherally or within the spinal cord area, is sufficient for long-term modulation of multiple types of nociception with hyperalgesic responses being most persistent.