RESUMEN
Increasing evidences demonstrate the role of sensory innervation in bone metabolism, remodeling and repair, however neurovascular coupling in bone is rarely studied. Using microfluidic devices as an indirect co-culture model to mimic in vitro the physiological scenario of innervation, our group demonstrated that sensory neurons (SNs) were able to regulate the extracellular matrix remodeling by endothelial cells (ECs), in particular through sensory neuropeptides, i.e. calcitonin gene-related peptide (CGRP) and substance P (SP). Nonetheless, still little is known about the cell signaling pathways and mechanism of action in neurovascular coupling. Here, in order to characterize the communication between SNs and ECs at molecular level, we evaluated the effect of SNs and the neuropeptides CGRP and SP on ECs. We focused on different pathways known to play a role on endothelial functions: calcium signaling, p38 and Erk1/2; the control of signal propagation through Cx43; and endothelial functions through the production of nitric oxide (NO). The effect of SNs was evaluated on ECs Ca2+ influx, the expression of Cx43, endothelial nitric oxide synthase (eNOS) and nitric oxide (NO) production, p38, ERK1/2 as well as their phosphorylated forms. In addition, the role of CGRP and SP were either analyzed using respective antagonists in the co-culture model, or by adding directly on the ECs monocultures. We show that capsaicin-stimulated SNs induce increased Ca2+ influx in ECs. SNs stimulate the increase of NO production in ECs, probably involving a decrease in the inhibitory eNOS T495 phosphorylation site. The neuropeptide CGRP, produced by SNs, seems to be one of the mediators of this effect in ECs since NO production is decreased in the presence of CGRP antagonist in the co-culture of ECs and SNs, and increased when ECs are stimulated with synthetic CGRP. Taken together, our results suggest that SNs play an important role in the control of the endothelial cell functions through CGRP production and NO signaling pathway.
Asunto(s)
Péptido Relacionado con Gen de Calcitonina , Células Endoteliales , Óxido Nítrico , Células Receptoras Sensoriales , Transducción de Señal , Sustancia P , Péptido Relacionado con Gen de Calcitonina/metabolismo , Péptido Relacionado con Gen de Calcitonina/farmacología , Sustancia P/farmacología , Sustancia P/metabolismo , Transducción de Señal/fisiología , Transducción de Señal/efectos de los fármacos , Células Endoteliales/efectos de los fármacos , Células Endoteliales/metabolismo , Células Receptoras Sensoriales/efectos de los fármacos , Células Receptoras Sensoriales/metabolismo , Animales , Óxido Nítrico/metabolismo , Técnicas de Cocultivo , Comunicación Celular/fisiología , Comunicación Celular/efectos de los fármacos , Óxido Nítrico Sintasa de Tipo III/metabolismo , Células Cultivadas , Humanos , RatasRESUMEN
The perineuronal net (PNN) is a well-described highly specialized extracellular matrix structure found in the central nervous system. Thus far, no reports of its presence or connection to pathological processes have been described in the peripheral nervous system. Our study demonstrates the presence of a PNN in the spinal afferent innervation of the distal colon of mice and characterizes structural and morphological alterations induced in an ulcerative colitis (UC) model. C57Bl/6 mice were given 3% dextran sulfate sodium (DSS) to induce acute or chronic UC. L6/S1 dorsal root ganglia (DRG) were collected. PNNs were labeled using fluorescein-conjugated Wisteria Floribunda (WFA) l lectin, and calcitonin gene-related peptide (CGRP) immunofluorescence was used to detect DRG neurons. Most DRG cell bodies and their extensions toward peripheral nerves were found surrounded by the PNN-like structure (WFA+), labeling neurons' cytoplasm and the pericellular surfaces. The amount of WFA+ neuronal cell bodies was increased in both acute and chronic UC, and the PNN-like structure around cell bodies was thicker in UC groups. In conclusion, a PNN-like structure around DRG neuronal cell bodies was described and found modulated by UC, as changes in quantity, morphology, and expression profile of the PNN were detected, suggesting a potential role in sensory neuron peripheral sensitization, possibly modulating the pain profile of ulcerative colitis.
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Colitis Ulcerosa , Colon , Ganglios Espinales , Ratones Endogámicos C57BL , Animales , Colitis Ulcerosa/patología , Colitis Ulcerosa/metabolismo , Ratones , Ganglios Espinales/patología , Ganglios Espinales/metabolismo , Colon/inervación , Colon/patología , Colon/metabolismo , Masculino , Péptido Relacionado con Gen de Calcitonina/metabolismo , Matriz Extracelular/patología , Matriz Extracelular/metabolismo , Sulfato de Dextran/toxicidad , Red Nerviosa/patología , Red Nerviosa/metabolismoRESUMEN
AIM: The purpose of this study was to quantify the effect of five different root canal preparation instruments on Substance P (SP), Calcitonin gene-related peptide (CGRP) and their receptors expression in healthy human periodontal ligament. METHODOLOGY: STROBE guidelines were used to design a study using 60 periodontal ligament samples obtained from healthy lower premolars where extraction was indicated for orthodontic reasons. Prior to extraction 40 of these premolars were equally divided into four groups and root canals were prepared using different systems: Mtwo, Reciproc Blue, HyFlex EDM and Plex-V. Ten premolars were prepared with hand files and served as a positive control group. The remaining 10 premolars where extracted without treatment and served as a negative control group. All periodontal ligament samples were processed to measure the expression of SP, CGRP and their receptors by radioimmunoassay. Kruskal-Wallis and Duncan tests were performed to determine statistically significant differences between the groups for each variable. RESULTS: Greater expression of all the peptides measured were found in the hand-file preparation group, followed by the Reciproc Blue, Mtwo, HyFlex EDM and Plex-V groups. The lower SP, CGRP and their receptors values were for the intact teeth control group. Kruskal-Wallis test showed statistically significant differences amongst groups (p < .001). Dunn post-hoc tests showed statistically significant differences in SP, CGRP and their receptors expression between the intact teeth and the hand-file and Reciproc Blue groups. Hand-file group showed significant differences with the other groups, except with Reciproc Blue, where no differences were observed in any of the peptides measured. Finally, no differences were observed between Plex-V and HyFlex in any of the peptides measured. CONCLUSIONS: Root canal preparation with hand files and Reciproc Blue generates the highest expression of SP, CGRP, NK1 and CGRP1R in human periodontal ligament, whilst Plex-V and HyFlex maintain the basal expression of neuropeptides and their receptors. Mtwo showed intermediate results between Reciproc Blue and HyFlex.
Asunto(s)
Péptido Relacionado con Gen de Calcitonina , Sustancia P , Humanos , Sustancia P/metabolismo , Péptido Relacionado con Gen de Calcitonina/metabolismo , Ligamento Periodontal/metabolismo , Preparación del Conducto Radicular , Diente Premolar , Cavidad Pulpar , Diseño de EquipoRESUMEN
BACKGROUND: Glomus Tumor (GT) are benign neoplasms that originate from mesenchymal cells. It presents as tenderness and cold allodynia in the digits, especially in the subungual region. There are a few studies that investigated the mechanism of pain. OBJECTIVES: To analyze the clinical-pathologic characteristics of GT and to identify the expression of IL-1ß, IL-6, and CGRP in it, further, to explore the possible mechanism of pain. METHODS: The clinical and pathological data of 60â¯GT patients were retrospectively analyzed. Tissue microarrays and immunohistochemistry were used to measure the expression of IL-1ß, IL-6 and CGRP. RESULTS: GT is more common in females and the ratio of male to was near to 1:2, mostly in middle-aged people. It often occurs in fingertips, especially the thumbs. Patients often present with spontaneous pain, tenderness, and cold hypersensitivity. Both the two pain mediators IL-1ß and IL-6 were highly expressed in GT cells of patients with and without cold hypersensitivity. While CGRP was not expressed in GT. STUDY LIMITATIONS: Low sample size and further research is needed to explore the specific mechanism. CONCLUSIONS: IL-1ß and IL-6 were highly expressed in GT cells, suggesting that IL-1ß and IL-6 have certain nociceptive roles in GT. In the 4 patients with cold intolerance, the intensity of IL-1ß and IL-6 staining was also strong, suggesting that they may not play a role in the cold hypersensitivity. However, since there are only 4 patients with cold intolerance, it's necessary to conduct further in-depth research using a larger sample size. The specific role of CGRP in GT has not been found yet.
Asunto(s)
Péptido Relacionado con Gen de Calcitonina , Síndromes Periódicos Asociados a Criopirina , Tumor Glómico , Persona de Mediana Edad , Femenino , Humanos , Masculino , Péptido Relacionado con Gen de Calcitonina/metabolismo , Tumor Glómico/patología , Estudios Retrospectivos , Interleucina-6 , Interleucina-1beta , DolorRESUMEN
Pain is a common non-motor symptom of Parkinson's disease (PD), which often occurs in the early disease stages. Despite the high prevalence, it remains inadequately treated. In a hemi-parkinsonian rat model, we aimed to investigate the neurochemical factors involved in orofacial pain development, with a specific focus on pain-related peptides and cannabinoid receptors. We also evaluated whether treadmill exercise could improve orofacial pain and modulate these mechanisms. Rats were unilaterally injected in the striatum with either 6-hydroxydopamine (6-OHDA) or saline. Fifteen days after stereotactic surgery, the animals were submitted to treadmill exercise (EX), or remained sedentary (SED). Pain assessment was performed before the surgical procedure and prior to each training session. Pain-related peptides, substance P (SP), calcitonin gene-related peptide (CGRP), and transient receptor potential vanilloid type 1 (TRPV1) activation and cannabinoid receptor type 1 (CB1) and type 2 (CB2) were evaluated in the trigeminal nucleus. In order to confirm the possible involvement of cannabinoid receptors, we also injected antagonists of CB1 and CB2 receptors. We confirmed the presence of orofacial pain after unilateral 6-OHDA-injection, which improved after aerobic exercise training. We also observed increased pain-related expression of SP, CGRP and TRPV1 and decreased CB1 and CB2 in the trigeminal ganglion and caudal spinal trigeminal nucleus in animals with PD, which was reversed after aerobic exercise training. In addition, we confirm the involvement of cannabinoid receptors since both antagonists decreased the nociceptive threshold of PD animals. These data suggest that aerobic exercise effectively improved the orofacial pain associated with the PD model, and may be mediated by pain-related neuropeptides and cannabinoid receptors in the trigeminal system.
Asunto(s)
Neuropéptidos , Enfermedad de Parkinson , Ratas , Animales , Enfermedad de Parkinson/complicaciones , Enfermedad de Parkinson/metabolismo , Péptido Relacionado con Gen de Calcitonina/metabolismo , Oxidopamina/toxicidad , Dolor Facial , Modelos Animales de EnfermedadRESUMEN
SUMMARY: Neuropeptide calcitonin gene-related peptide (CGRP) is a neurotransmitter related to vasculogenesis during organ development. The vascular endothelial growth factor A (VEGF-A) is also required for vascular patterning during lung morphogenesis. CGRP is primarily found in organs and initially appears in pulmonary neuroendocrine cells during the early embryonic stage of lung development. However, the relationship between CGRP and VEGF-A during lung formation remains unclear. This study investigates CGRP and VEGF-A mRNA expressions in the embryonic, pseudoglandular, canalicular, saccular, and alveolar stages of lung development from embryonic day 12.5 (E12.5) to postnatal day 5 (P5) through quantitative real-time polymerase chain reaction (qRT-PCR) and in situ hybridization. Further, we analyzed the expression of CGRP via immunohistochemistry. The VEGF-A mRNA was mainly scattered across the whole lung body from E12.5. CGRP was found to be expressed in a few epithelial cells of the canalicular and the respiratory bronchiole of the lung from E12.5 to P5. An antisense probe for CGRP mRNA was strongly detected in the lung from E14.5 to E17.5. Endogenous CGRP may regulate the development of the embryonic alveoli from E14.5 to E17.5 in a temporal manner.
El péptido relacionado con el gen de la calcitonina (CGRP) es un neurotransmisor vinculado con la vasculogénesis durante el desarrollo de órganos. El factor de crecimiento endotelial vascular A (VEGF-A) también se requiere para el patrón vascular durante la morfogénesis pulmonar. El CGRP se encuentra principalmente en los órganos y aparece inicialmente en las células neuroendocrinas pulmonares durante la etapa embrionaria temprana del desarrollo pulmonar. Sin embargo, la relación entre CGRP y VEGF-A durante la formación de los pulmones sigue sin estar clara. Este estudio investiga las expresiones de ARNm de CGRP y VEGF-A en las etapas embrionaria, pseudoglandular, canalicular, sacular y alveolar del desarrollo pulmonar desde el día embrionario 12,5 (E12,5) hasta el día postnatal 5 (P5) a través de la reacción en cadena de la polimerasa cuantitativa en tiempo real. (qRT-PCR) e hibridación in situ. Además, analizamos la expresión de CGRP mediante inmunohistoquímica. El ARNm de VEGF-A se dispersó principalmente por todo parénquima pulmonar desde E12,5. Se encontró que CGRP se expresaba en unas pocas células epiteliales de los bronquiolos canaliculares y respiratorios del pulmón desde E12,5 a P5. Se detectó fuertemente una sonda antisentido para ARNm de CGRP en el pulmón de E14,5 a E17,5. El CGRP endógeno puede regular el desarrollo de los alvéolos embrionarios de E14,5 a E17,5 de manera temporal.
Asunto(s)
Animales , Ratones , Péptido Relacionado con Gen de Calcitonina/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismo , Pulmón/crecimiento & desarrollo , Pulmón/embriología , Inmunohistoquímica , Hibridación in Situ , Neurotransmisores , Neovascularización FisiológicaRESUMEN
Previous studies have reported that L5/L6 spinal nerve ligation (SNL), but not L5 spinal nerve transection (SNT), enhances anoctamin-1 in injured and uninjured dorsal root ganglia (DRG) of rats suggesting some differences in function of the type of nerve injury. The role of bestrophin-1 in these conditions is unknown. The aim of this study was to investigate the role of bestrophin-1 in rats subjected to L5 SNT and L5/L6 SNL. SNT up-regulated bestrophin-1 protein expression in injured L5 and uninjured L4 DRG at day 7, whereas it enhanced GAP43 mainly in injured, but also in uninjured DRG. In contrast, SNL enhanced GAP43 at day 1 and 7, while bestrophin-1 expression increased only at day 1 after nerve injury. Accordingly, intrathecal injection of the bestrophin-1 blocker CaCCinh-A01 (1-10 µg) reverted SNT- or SNL-induced tactile allodynia in a concentration-dependent manner. Intrathecal injection of CaCCinh-A01 (10 µg) prevented SNT-induced upregulation of bestrophin-1 and GAP43 at day 7. In contrast, CaCCinh-A01 did not affect SNL-induced up-regulation of GAP43 nor bestrophin-1. Bestrophin-1 was mainly expressed in small- and medium-size neurons in naïve rats, while SNT increased bestrophin-1 immunoreactivity in CGRP+, but not in IB4+ neuronal cells in DRG. Intrathecal injection of bestrophin-1 plasmid (pCMVBest) induced tactile allodynia and increased bestrophin-1 expression in DRG and spinal cord in naïve rats. CaCCinh-A01 reversed bestrophin-1 overexpression-induced tactile allodynia and restored bestrophin-1 expression. Our data suggest that bestrophin-1 plays a relevant role in neuropathic pain induced by SNT, but not by SNL. PERSPECTIVE: SNT, but not SNL, up-regulates bestrophin-1 and GAP43 protein expression in injured L5 and uninjured L4 DRG. SNT increases bestrophin-1 immunoreactivity in CGRP+ neurons in DRG. Bestrophin-1 overexpression induces allodynia. CaCCinh-A01 reduces allodynia and restores bestrophin-1 expression. Our data suggest bestrophin-1 is differentially regulated depending on the neuropathic pain model.
Asunto(s)
Hiperalgesia , Neuralgia , Ratas , Animales , Bestrofinas/metabolismo , Hiperalgesia/metabolismo , Ratas Sprague-Dawley , Péptido Relacionado con Gen de Calcitonina/metabolismo , Neuralgia/metabolismo , Nervios Espinales/lesiones , Ligadura , Canales de Cloruro/metabolismo , Ganglios Espinales/metabolismoRESUMEN
ABSTRACT: The loss of GABAergic inhibition is a mechanism that underlies neuropathic pain. Therefore, rescuing the GABAergic inhibitory tone through the activation of GABA A receptors is a strategy to reduce neuropathic pain. This study was designed to elucidate the function of the spinal α 6 -containing GABA A receptor in physiological conditions and neuropathic pain in female and male rats. Results show that α 6 -containing GABA A receptor blockade or transient α 6 -containing GABA A receptor knockdown induces evoked hypersensitivity and spontaneous pain in naive female rats. The α 6 subunit is expressed in IB4 + and CGRP + primary afferent neurons in the rat spinal dorsal horn and dorsal root ganglia but not astrocytes. Nerve injury reduces α 6 subunit protein expression in the central terminals of the primary afferent neurons and dorsal root ganglia, whereas intrathecal administration of positive allosteric modulators of the α 6 -containing GABA A receptor reduces tactile allodynia and spontaneous nociceptive behaviors in female, but not male, neuropathic rats and mice. Overexpression of the spinal α 6 subunit reduces tactile allodynia and restores α 6 subunit expression in neuropathic rats. Positive allosteric modulators of the α 6 -containing GABA A receptor induces a greater antiallodynic effect in female rats and mice compared with male rats and mice. Finally, α 6 subunit is expressed in humans. This receptor is found in CGRP + and P2X3 + primary afferent fibers but not astrocytes in the human spinal dorsal horn. Our results suggest that the spinal α 6 -containing GABA A receptor has a sex-specific antinociceptive role in neuropathic pain, suggesting that this receptor may represent an interesting target to develop a novel treatment for neuropathic pain.
Asunto(s)
Neuralgia , Receptores de GABA-A , Masculino , Ratas , Femenino , Ratones , Humanos , Animales , Receptores de GABA-A/metabolismo , Hiperalgesia , Péptido Relacionado con Gen de Calcitonina/metabolismo , Asta Dorsal de la Médula Espinal/metabolismoRESUMEN
Blood pressure is a highly controlled cardiovascular parameter that normally guarantees an adequate blood supply to all body tissues. This parameter is mainly regulated by peripheral vascular resistance and is maintained by local mediators (i.e., autacoids), and by the nervous and endocrine systems. Regarding the nervous system, blood pressure can be modulated at the central level by regulating the autonomic output. However, at peripheral level, there exists a modulation by activation of prejunctional monoaminergic receptors in autonomic- or sensory-perivascular fibers. These modulatory mechanisms on resistance blood vessels exert an effect on the release of neuroactive substances from the autonomic or sensory fibers that modify blood pressure. Certainly, resistance blood vessels are innervated by perivascular: (i) autonomic sympathetic fibers (producing vasoconstriction mainly by noradrenaline release); and (ii) peptidergic sensory fibers [producing vasodilatation mainly by calcitonin gene-related peptide (CGRP) release]. In the last years, by using pithed rats, several monoaminergic mechanisms for controlling both the sympathetic and sensory perivascular outflows have been elucidated. Additionally, several studies have shown the functions of many monoaminergic auto-receptors and hetero-receptors expressed on perivascular fibers that modulate neurotransmitter release. On this basis, the present review: (i) summarizes the modulation of the peripheral vascular tone by adrenergic, serotoninergic, dopaminergic, and histaminergic receptors on perivascular autonomic (sympathetic) and sensory fibers, and (ii) highlights that these monoaminergic receptors are potential therapeutic targets for the development of novel medications to treat cardiovascular diseases (with some of them explored in clinical trials or already in clinical use).
Asunto(s)
Sistema Nervioso Autónomo/metabolismo , Monoaminas Biogénicas/metabolismo , Péptido Relacionado con Gen de Calcitonina/metabolismo , Receptores de Amina Biogénica/metabolismo , Fibras Adrenérgicas/metabolismo , Animales , Presión Sanguínea/fisiología , Encéfalo/metabolismo , Humanos , Receptores Adrenérgicos/metabolismoRESUMEN
OBJECTIVES: The purpose of this study was to evaluate the relationship between the serum levels of calcitonin gene-related peptide (CGRP) and the prognosis of pediatric patients with severe pneumonia. METHODS: Children diagnosed with severe pneumonia (n=76) were stratified into the survival (n=58) and non-survival groups (n=18) according to their 28-day survival status and into the non-risk (n=51), risk (n=17) and high-risk (n=8) categories based on the pediatric critical illness score (PCIS). Demographic data and laboratory results were collected. Serum CGRP levels were determined by enzyme-linked immunosorbent assay (ELISA). A receiver operating characteristic (ROC) curve was generated to determine the cutoff score for high CGRP levels. RESULTS: Serum CGRP levels were significantly higher in the survival group than in the non-survival group and were significantly higher in the non-risk group than in the risk and high-risk groups. The ROC curve for the prognostic potential of CGRP yielded a significant area under the curve (AUC) value with considerable sensitivity and specificity. CONCLUSION: Our findings show that CGRP downregulation might be a diagnostic marker that predicts the prognosis and survival of children with severe pneumonia.
Asunto(s)
Péptido Relacionado con Gen de Calcitonina/genética , Neumonía/sangre , Precursores de Proteínas/sangre , Vasodilatadores/metabolismo , Calcitonina , Péptido Relacionado con Gen de Calcitonina/sangre , Péptido Relacionado con Gen de Calcitonina/metabolismo , Niño , Enfermedad Crítica , Femenino , Humanos , Masculino , Neumonía/mortalidad , Pronóstico , Curva ROC , Análisis de SupervivenciaRESUMEN
This study analyzed the effects of photobiomodulation (PBM) with low-level laser therapy on nociceptive behavior and neuronal activity in the trigeminal nucleus after experimental unilateral temporomandibular joint (TMJ) disc injury. The animals were divided into 4 groups (n = 10 each): group 1, surgical injury of the articular disc and PBM; group 2, sham-operated subjected to PBM; group 3, surgical injury of the articular disc; and group 4, control (Naïve). Ten sessions of PBM were performed using GaAs laser with a wavelength of 904 nm, power of 75 W pico, average power of 0.043 W, area of the beam of 0.13 cm2, duration of the pulses of 60 nseg (in the frequency of 9500 Hz), energy density of 5.95 J/cm2, energy per point of 0.7 J, and power density of 333.8 mW/cm2, and the irradiation was done for 18 s per point. Neuropathic symptoms were evaluated using the von Frey test. Trigeminal ganglion samples underwent immunoblotting to examine the expression of substance P, vanilloid transient potential receptor of subtype-1 (TRPV-1), and peptide related to the calcitonin gene (CGRP). There was a total decrease in pain sensitivity after the second session of PBM in operated animals, and this decrease remains until the last session. There was a significant decrease in the expression of SP, TRPV-1, and CGRP after PBM. Photobiomodulation therapy was effective in reducing nociceptive behavior and trigeminal nucleus neuronal activity after TMJ disc injury.
Asunto(s)
Terapia por Luz de Baja Intensidad , Neuropéptidos/metabolismo , Umbral del Dolor , Dolor/radioterapia , Articulación Temporomandibular/patología , Articulación Temporomandibular/efectos de la radiación , Animales , Conducta Animal , Péptido Relacionado con Gen de Calcitonina/metabolismo , Láseres de Semiconductores , Masculino , Ratas Wistar , Sustancia P/metabolismo , Canales Catiónicos TRPV/metabolismo , Resultado del TratamientoRESUMEN
Lichen planopilaris (LPP) and frontal fibrosing alopecia (FFA) are lymphocytic scarring alopecias affecting primarily the scalp. Although both diseases may share some clinical and histopathological features, in the last decade, FFA has become an "epidemic" particularly in Europe, North and South America with unique clinical manifestations compared to LPP, thus, raising the idea that this disease may have a different pathogenesis. Symptoms such as scalp burning, pruritus or pain are usually present in both diseases, suggesting a possible role for nerves and neuropeptides in the pathogenesis of both diseases. Based on some previous studies, neuropeptides, such as substance P (SP) and calcitonin gene-related peptide (CGRP), have been associated with lipid metabolism and many chronic inflammatory disorders. In this study, we asked if these neuropeptides are associated with LPP and FFA scalp lesions. Alteration in the expression of SP and CGRP in affected and unaffected scalp skin from patients with both diseases was found with examination of sections using immunohistochemical techniques and confocal microscopy. We then quantitatively assessed and compared SP and CGRP expression from control, LPP and FFA scalp biopsies. Although LPP and FFA share similar histopathologic findings, opposite results were found in affected and unaffected scalp in the ELISA tests, suggesting that these diseases may have different pathogenic mechanisms. We also found presence of histopathological inflammation irrespective of evident clinical lesions, which raises the possibility that both diseases may be more generalized processes affecting the scalp.
Asunto(s)
Alopecia/patología , Liquen Plano/fisiopatología , Inflamación Neurogénica/patología , Adulto , Anciano , Anciano de 80 o más Años , Biopsia , Péptido Relacionado con Gen de Calcitonina/metabolismo , Enfermedad Crónica , Epidermis/metabolismo , Femenino , Humanos , Inmunohistoquímica , Inflamación , Metabolismo de los Lípidos , Linfocitos/patología , Masculino , Microscopía Confocal , Persona de Mediana Edad , Neuropéptidos/química , Cuero Cabelludo/patología , Dermatosis del Cuero Cabelludo/patología , Sustancia P/metabolismoRESUMEN
OBJECTIVE: To evaluate whether intraganglionic calcitonin gene-related peptide induced differential migraine-like responses in male and female rats. METHODS: Calcitonin gene-related peptide was injected in the trigeminal ganglion of male and female rats followed by assessment of periorbital mechanical allodynia with von Frey hairs. The influence of systemic treatment with sumatriptan or intraganglionic treatment with minocycline and propentofylline was determined on the calcitonin gene-related peptide-induced mechanical allodynia in male and female rats. One additional group was exposed to an aversive light 24 h after calcitonin gene-related peptide priming, followed by evaluation of periorbital mechanical threshold, and another group was tested in the elevated-plus maze. RESULTS: Intraganglionar calcitonin gene-related peptide-induced periorbital mechanical allodynia in female (0.5 to 6 h) and male rats (0.5 to 4 h). Systemic sumatriptan briefly attenuated the mechanical allodynia, but intraganglionar minocycline or propentofylline injection was effective only in male rats. Calcitonin gene-related peptide induced photic sensitivity in female and male rats (lasting 4 h and 1 h, respectively), as well as anxiety-like behavior. CONCLUSIONS: Intraganglionar calcitonin gene-related peptide may play a major role in migraine-like responses, including periorbital mechanical allodynia, light sensitivity and anxiety like-behavior. Female rats are likely to be more susceptible to calcitonin gene-related peptide effects and a better understanding of the sexual dimorphism in calcitonin gene-related peptide signaling may help to improve migraine therapy.
Asunto(s)
Péptido Relacionado con Gen de Calcitonina/metabolismo , Hiperalgesia/metabolismo , Trastornos Migrañosos/metabolismo , Ganglio del Trigémino/metabolismo , Animales , Péptido Relacionado con Gen de Calcitonina/farmacología , Femenino , Hiperalgesia/inducido químicamente , Masculino , Trastornos Migrañosos/inducido químicamente , Ratas , Ratas Wistar , Agonistas del Receptor de Serotonina 5-HT1/farmacología , Caracteres Sexuales , Sumatriptán/farmacología , Ganglio del Trigémino/efectos de los fármacosRESUMEN
OBJECTIVES: The purpose of this study was to evaluate the relationship between the serum levels of calcitonin gene-related peptide (CGRP) and the prognosis of pediatric patients with severe pneumonia. METHODS: Children diagnosed with severe pneumonia (n=76) were stratified into the survival (n=58) and non-survival groups (n=18) according to their 28-day survival status and into the non-risk (n=51), risk (n=17) and high-risk (n=8) categories based on the pediatric critical illness score (PCIS). Demographic data and laboratory results were collected. Serum CGRP levels were determined by enzyme-linked immunosorbent assay (ELISA). A receiver operating characteristic (ROC) curve was generated to determine the cutoff score for high CGRP levels. RESULTS: Serum CGRP levels were significantly higher in the survival group than in the non-survival group and were significantly higher in the non-risk group than in the risk and high-risk groups. The ROC curve for the prognostic potential of CGRP yielded a significant area under the curve (AUC) value with considerable sensitivity and specificity. CONCLUSION: Our findings show that CGRP downregulation might be a diagnostic marker that predicts the prognosis and survival of children with severe pneumonia.
Asunto(s)
Humanos , Masculino , Femenino , Niño , Neumonía/sangre , Precursores de Proteínas/sangre , Vasodilatadores/metabolismo , Péptido Relacionado con Gen de Calcitonina/genética , Neumonía/mortalidad , Pronóstico , Calcitonina , Péptido Relacionado con Gen de Calcitonina/metabolismo , Péptido Relacionado con Gen de Calcitonina/sangre , Análisis de Supervivencia , Curva ROC , Enfermedad CríticaRESUMEN
The sensory nervous system controls cardiovascular homeostasis via capsaicin-sensitive neurons that release calcitonin gene-related peptide (CGRP), which subsequently activates CGRP receptors. How this perivascular CGRPergic discharge is modulated, nevertheless, remains unclear. In pithed rats, systemic vasodilation induced by CGRPergic discharge stimulation results in diastolic blood pressure (BP) decrements that are inhibited by the dopamine D2-like receptor agonist quinpirole. Since this inhibition is mediated by raclopride- or haloperidol-sensitive D2-like receptors (comprising the D2, D3, and D4 subtypes), the present study pharmacologically investigated the specific contribution of these subtypes to the modulation of the systemic CGRPergic vasodilation, using highly specific antagonists. To that end, 55 male Wistar rats were pithed for thoracic (T9-T12) spinal stimulation of the perivascular CGRPergic discharge. The resulting frequency-dependent decrements in diastolic BP were inhibited by quinpirole, and this sensory-inhibition was (a) unchanged after i.v. injections of the antagonists L-741,626 (D2) or L-745,870 (D4) and (b) completely blocked by SB-277011-A (D3). Accordingly, we suggest the main role of the D3 receptor subtypes in the inhibition by quinpirole of the neurogenic CGRPergic systemic vasodilation. These findings contribute to a better understanding of the dopaminergic modulation of the rat perivascular CGRPergic discharge producing systemic vasodilation.
Asunto(s)
Presión Sanguínea/efectos de los fármacos , Péptido Relacionado con Gen de Calcitonina/metabolismo , Agonistas de Dopamina/farmacología , Frecuencia Cardíaca/fisiología , Receptores de Dopamina D3/antagonistas & inhibidores , Vasodilatación/efectos de los fármacos , Animales , Estimulación Eléctrica , Frecuencia Cardíaca/efectos de los fármacos , Masculino , Quinpirol/farmacología , Racloprida/farmacología , Ratas , Ratas WistarRESUMEN
Blood flow distribution relies on precise coordinated control of vasomotor tone of resistance arteries by complex signalling interactions between perivascular nerves and endothelial cells. Sympathetic nerves are vasoconstrictors, whereas endothelium-dependent NO production provides a vasodilator component. In addition, resistance vessels are also innervated by sensory nerves, which are activated during inflammation and cause vasodilation by the release of calcitonin gene-related peptide (CGRP). Inflammation leads to superoxide anion (O2⢠-) formation and endothelial dysfunction, but the involvement of CGRP in this process has not been evaluated. Here we show a novel mechanistic relation between perivascular sensory nerve-derived CGRP and the development of endothelial dysfunction. CGRP receptor stimulation leads to pannexin-1-formed channel opening and the subsequent O2⢠--dependent connexin-based hemichannel activation in endothelial cells. The prolonged opening of these channels results in a progressive inhibition of NO production. These findings provide new therapeutic targets for the treatment of the inflammation-initiated endothelial dysfunction.
Asunto(s)
Péptido Relacionado con Gen de Calcitonina/metabolismo , Conexinas/metabolismo , Células Endoteliales/metabolismo , Inflamación/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Óxido Nítrico/metabolismo , Animales , Células Endoteliales/patología , Inflamación/patología , Masculino , Ratas Sprague-Dawley , Transducción de Señal , Superóxidos/metabolismoRESUMEN
BACKGROUND AND PURPOSE: Maresin 1 (MaR1) is a specialised pro-resolving lipid mediator with anti-inflammatory and analgesic activities. In this study, we addressed the modulation of peripheral and spinal cord cells by MaR1 in the context of inflammatory pain. EXPERIMENTAL APPROACH: Mice were treated with MaR1 before intraplantar injection of carrageenan or complete Freund's adjuvant (CFA). Mechanical hyperalgesia was assessed using the electronic von Frey and thermal hyperalgesia using a hot plate. Spinal cytokine production and NF-κB activation were determined by ELISA and astrocytes and microglia activation by RT-qPCR and immunofluorescence. CGRP release by dorsal root ganglia (DRG) neurons was determined by EIA. Neutrophil and macrophage recruitment were determined by immunofluorescence, flow cytometry, and colorimetric methods. Trpv1 and Nav1.8 expression and calcium imaging of DRG neurons were determined by RT-qPCR and Fluo-4AM respectively. KEY RESULTS: MaR1 reduced carrageenan- and CFA-induced mechanical and thermal hyperalgesia and neutrophil and macrophage recruitment proximal to CGRP+ fibres in the paw skin. Moreover, MaR1 reduced NF-κB activation, IL-1ß and TNF-α production, and spinal cord glial cells activation. In the DRG, MaR1 reduced CFA-induced Nav1.8 and Trpv1 mRNA expression and calcium influx and capsaicin-induced release of CGRP by DRG neurons. CONCLUSIONS AND IMPLICATIONS: MaR1 reduced DRG neurons activation and CGRP release explaining, at least in part, its analgesic and anti-inflammatory effects. The enduring analgesic and anti-inflammatory effects and also post-treatment activity of MaR1 suggest that specialised pro-resolving lipid mediators have potential as a new class of drugs for the treatment of inflammatory pain.
Asunto(s)
Analgésicos/uso terapéutico , Antiinflamatorios/uso terapéutico , Ácidos Docosahexaenoicos/uso terapéutico , Hiperalgesia/tratamiento farmacológico , Dolor/tratamiento farmacológico , Analgésicos/farmacología , Animales , Antiinflamatorios/farmacología , Péptido Relacionado con Gen de Calcitonina/metabolismo , Carragenina , Ácidos Docosahexaenoicos/farmacología , Adyuvante de Freund , Ganglios Espinales/efectos de los fármacos , Calor , Hiperalgesia/genética , Hiperalgesia/metabolismo , Interleucina-1beta/metabolismo , Masculino , Ratones , Canal de Sodio Activado por Voltaje NAV1.8/genética , FN-kappa B/metabolismo , Neuronas/efectos de los fármacos , Dolor/genética , Dolor/metabolismo , Estimulación Física , Médula Espinal/efectos de los fármacos , Médula Espinal/metabolismo , Canales Catiónicos TRPV/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
The aim of this study was to investigate the effect of a placebo, intracanal diode laser application, and low-level laser therapy (LLLT) on the change of the total amount of calcitonin gene-related peptide (CGRP) in the gingival crevicular fluid (GCF) (split-mouth study design). GCF sampling was performed on a contralateral tooth and experimental tooth (root canal-treated tooth) of thirty-nine patients. The patients were divided into three groups (n = 13), as follows: placebo (mock laser application), intracanal laser application, and LLLT. GCF sampling was repeated at the same sites (experimental and control teeth) one week after root canal treatment. The data were analyzed using the Pearson's correlation analysis and the independent-samples t-tests (p=0.05). In the placebo group, the total CGRP level changes in the GCF before and after treatment was significantly higher for experimental teeth than for control teeth (p<0.05). However, there was no significant difference between experimental and control teeth in the intracanal laser application and LLLT groups (p > 0.05). Intracanal laser application and low-level laser therapy have immunomodulation effects linked to the modulation of the total amount of CGRP in the GCF.
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Péptido Relacionado con Gen de Calcitonina/efectos de la radiación , Terapia por Luz de Baja Intensidad/métodos , Tratamiento del Conducto Radicular/métodos , Adulto , Péptido Relacionado con Gen de Calcitonina/metabolismo , Estudios de Casos y Controles , Femenino , Líquido del Surco Gingival/efectos de la radiación , Humanos , Láseres de Semiconductores , Masculino , Resultado del TratamientoRESUMEN
This study aimed to evaluate neuropeptide expression after bleaching treatment using histopathological and immunohistochemical analyses and the effects of hydrocortisone and acetaminophen on pulp inflammation, sine dental bleaching and inflammation first occur, and only then, the treatmentt. Sixty-three rats were divided into three groups (n = 21) according to the pain-relieving therapy used: I-control; II-topical application of Otosporin for 10 min after the bleaching treatment; III-oral administration of paracetamol 30 min before whitening and then every 12h. In all the study groups, placebo gel was applied to the left upper jaw (control) and a 35% H2O2-based whitening gel was applied to the right upper jaw for 45 min. Seven animals from each group were euthanized at different time points: 0h after treatment, 24h, and 48h. After euthanasia, the first molar on each side was analyzed by histology and immunohistochemistry to assess the degree of inflammation and verify the presence of the neuropeptides, substance P (SP) and calcitonin gene-related peptide (CGRP). The data were analyzed using the statistical nonparametric Kruskal-Wallis test followed by Dunn's test for individual comparisons. Extensive areas of necrosis were observed in the groups that received bleaching treatment only, whereas reduced damage were obtained in the group treated with Otosporin. The immunohistochemical analysis showed positive immunolabeling in all groups, including the control, but this was stronger in the groups that received bleaching treatment. The best results were obtained in the group that received treatment with Otosporin. The use of Otosporin after dental bleaching minimized the side effects of this treatment.
Asunto(s)
Antiinflamatorios/uso terapéutico , Pulpa Dental/efectos de los fármacos , Pulpa Dental/patología , Pulpitis/tratamiento farmacológico , Pulpitis/etiología , Blanqueamiento de Dientes/efectos adversos , Acetaminofén/uso terapéutico , Animales , Péptido Relacionado con Gen de Calcitonina/metabolismo , Pulpa Dental/metabolismo , Modelos Animales de Enfermedad , Combinación de Medicamentos , Humanos , Hidrocortisona/uso terapéutico , Inmunohistoquímica , Masculino , Neomicina/uso terapéutico , Polimixina B/uso terapéutico , Pulpitis/patología , Ratas , Ratas Wistar , Sustancia P/metabolismo , Blanqueadores Dentales/efectos adversosRESUMEN
Transcription factors are proteins that modulate the transcriptional rate of target genes in the nucleus in response to extracellular or cytoplasmic signals. Activating transcription factors 2 (ATF2) and 3 (ATF3) respond to environmental signals and maintain cellular homeostasis. There is evidence that inflammation and nerve injury modulate ATF2 and ATF3 expression. However, the function of these transcription factors in pain is unknown. The purpose of this study was to investigate the contribution of ATF2 and ATF3 to nerve injury-induced neuropathic pain. L5/6 spinal nerve ligation induced tactile allodynia and thermal hyperalgesia. Moreover, nerve damage enhanced ATF2 and ATF3 protein expression in injured L5/6 dorsal root ganglia and spinal cord but not in uninjured L4 dorsal root ganglia. Nerve damage also enhanced ATF2 immunoreactivity in dorsal root ganglia and spinal cord 7 to 21 days post-injury. Repeated intrathecal post-treatment with a small-interfering RNA targeted against ATF2 (ATF2 siRNA) or anti-ATF2 antibody partially reversed tactile allodynia and thermal hyperalgesia. In contrast, ATF3 siRNA or anti-ATF3 antibody did not modify nociceptive behaviors. ATF2 immunoreactivity was found in dorsal root ganglia and spinal cord co-labeling with NeuN mainly in non-peptidergic (IB4+) but also in peptidergic (CGRP+) neurons. ATF2 was found mainly in small- and medium-sized neurons. These results suggest that ATF2, but not ATF3, is found in strategic sites related to spinal nociceptive processing and participates in the maintenance of neuropathic pain in rats.