RESUMEN
After spinal cord injury (SCI), blood-spinal cord barrier (BSCB) disruption and progressive hemorrhage lead to secondary injury, subsequent apoptosis and/or necrosis of neurons and glia, causing permanent neurological deficits. Growing evidence indicates that mithramycin A (MA), an anti-cancer drug, has neuroprotective effects in ischemic brain injury and Huntington's disease (HD). However, the precise mechanism underlying its protective effects is largely unknown. Here, we examined the effect of MA on BSCB breakdown and hemorrhage as well as subsequent inflammation after SCI. After moderate spinal cord contusion injury at T9, MA (150 µg/kg) was immediately injected intraperitoneally (i.p.) and further injected once a day for 5 days. Our data show that MA attenuated BSCB disruption and hemorrhage, and inhibited the infiltration of neutrophils and macrophages after SCI. Consistent with these findings, the expression of inflammatory mediators was significantly alleviated by MA. MA also inhibited the expression and activation of matrix metalloprotease-9 (MMP-9) after injury, which is known to disrupt BSCB and the degradation of tight junction (TJ) proteins. In addition, the expression of sulfonylurea receptor 1 (SUR1) and transient receptor potential melastatin 4 (TRPM4), which are known to mediate hemorrhage at an early stage after SCI, was significantly blocked by MA treatment. Finally, MA inhibited apoptotic cell death and improved functional recovery after injury. Thus, our results demonstrated that MA improves functional recovery by attenuating BSCB disruption and hemorrhage through the downregulation of SUR1/TRPM4 and MMP-9 after SCI.
Asunto(s)
Permeabilidad Capilar/efectos de los fármacos , Fármacos Neuroprotectores/farmacología , Plicamicina/análogos & derivados , Recuperación de la Función/efectos de los fármacos , Traumatismos de la Médula Espinal/patología , Animales , Hemorragia/patología , Masculino , Metaloproteinasa 9 de la Matriz/biosíntesis , Metaloproteinasa 9 de la Matriz/efectos de los fármacos , Plicamicina/farmacología , Ratas , Ratas Sprague-Dawley , Receptores de Sulfonilureas/biosíntesis , Receptores de Sulfonilureas/efectos de los fármacos , Canales Catiónicos TRPM/biosíntesis , Canales Catiónicos TRPM/efectos de los fármacosRESUMEN
The disruption of the blood-spinal cord barrier (BSCB) by matrix metalloprotease (MMP) activation is a detrimental event that leads to blood cell infiltration, inflammation, and apoptosis, thereby contributing to permanent neurological disability after spinal cord injury (SCI). However, the molecular mechanisms underlying Mmp gene regulation have not been fully elucidated. Here, we demonstrated the critical role of histone H3K27 demethylase Jmjd3 in the regulation of Mmp gene expression and BSCB disruption using in vitro cellular and in vivo animal models. We found that Jmjd3 up-regulation, in cooperation with NF-κB, after SCI is required for Mmp-3 and Mmp-9 gene expressions in injured vascular endothelial cells. In addition, Jmjd3 mRNA depletion inhibited Mmp-3 and Mmp-9 gene expressions and significantly attenuated BSCB permeability and the loss of tight junction proteins. These events further led to improved functional recovery, along with decreased hemorrhage, blood cell infiltration, inflammation, and cell death of neurons and oligodendrocytes after SCI. Thus, our findings suggest that Jmjd3 regulation may serve as a potential therapeutic intervention for preserving BSCB integrity following SCI.
Asunto(s)
Regulación de la Expresión Génica/genética , Histona Demetilasas con Dominio de Jumonji/metabolismo , Metaloproteinasa 3 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Traumatismos de la Médula Espinal/metabolismo , Traumatismos de la Médula Espinal/fisiopatología , Animales , Barrera Hematoencefálica , Permeabilidad Capilar/genética , Células Endoteliales/metabolismo , Masculino , FN-kappa B/metabolismo , Ratas Sprague-Dawley , Recuperación de la Función/fisiología , Médula Espinal/metabolismo , Regulación hacia ArribaRESUMEN
Recently we reported that fluoxetine (10 mg/kg) improves functional recovery by attenuating blood spinal cord barrier (BSCB) disruption after spinal cord injury (SCI). Here we investigated whether a low-dose of fluoxetine (1 mg/kg) and vitamin C (100 mg/kg), separately not possessing any protective effect, prevents BSCB disruption and improves functional recovery when combined. After a moderate contusion injury at T9 in rat, a low-dose of fluoxetine and vitamin C, or the combination of both was administered intraperitoneally immediately after SCI and further treated once a day for 14 d. Co-treatment with fluoxetine and vitamin C significantly attenuated BSCB permeability at 1 d after SCI. When only fluoxetine or vitamin C was treated after injury, however, there was no effect on BSCB disruption. Co-treatment with fluoxetine and vitamin C also significantly inhibited the expression and activation of MMP-9 at 8 h and 1 d after injury, respectively, and the infiltration of neutrophils (at 1 d) and macrophages (at 5 d) and the expression of inflammatory mediators (at 2 h, 6 h, 8 h or 24 h after injury) were significantly inhibited by co-treatment with fluoxetine and vitamin C. Furthermore, the combination of fluoxetine and vitamin C attenuated apoptotic cell death at 1 d and 5 d and improved locomotor function at 5 weeks after SCI. These results demonstrate the synergistic effect combination of low-dose fluoxetine and vitamin C on BSCB disruption after SCI and furthermore support the effectiveness of the combination treatment regimen for the management of acute SCI.
Asunto(s)
Ácido Ascórbico/administración & dosificación , Barrera Hematoencefálica/efectos de los fármacos , Fluoxetina/administración & dosificación , Recuperación de la Función/efectos de los fármacos , Traumatismos de la Médula Espinal/tratamiento farmacológico , Médula Espinal/efectos de los fármacos , Animales , Antioxidantes/administración & dosificación , Barrera Hematoencefálica/metabolismo , Relación Dosis-Respuesta a Droga , Sinergismo Farmacológico , Quimioterapia Combinada , Masculino , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Recuperación de la Función/fisiología , Inhibidores Selectivos de la Recaptación de Serotonina/administración & dosificación , Médula Espinal/metabolismo , Traumatismos de la Médula Espinal/metabolismo , Traumatismos de la Médula Espinal/fisiopatologíaRESUMEN
The activation of microglia after spinal cord injury (SCI) contributes to secondary damage by producing pro-inflammatory cytokines and mediators, leading to cell death of oligodendrocytes and neurons. Here, we show that matrix metalloprotease-3 (MMP-3) produced and secreted in the endothelial cells of blood vessels after SCI mediates microglial activation. MMP-3 was produced and secreted in bEnd.3 cells, a mouse brain-derived endothelial cell line, by oxygen-glucose deprivation/reoxygenation (OGD/RO). OGD/RO-induced MMP-3 expression and activity was also significantly inhibited by ghrelin, which was dependent on the ghrelin receptor GHS-R1a. Furthermore, the secreted MMP-3 from OGD/RO-induced bEnd.3 cells activated BV-2 cells, a murine microglial cell line. We also found that microglial activation after SCI was attenuated in MMP-3 knockout (KO) mice compared with wild type (WT) mice. Both p38 mitogen-activated protein kinase (MAPK) activation and pro-nerve growth factor (proNGF) production were more inhibited in MMP-3 KO than WT mice at 5d after injury. When WT mice were treated with Mmp-3 siRNA after injury, MMP-3 activity, microglial activation, p38MAPK activation and proNGF expression were significantly inhibited. Ghrelin treatment also significantly inhibited MMP-3 expression and activation after SCI, which was dependent on GHS-R1a. Finally, RhoA activation and oligodendrocyte cell death after injury were attenuated by Mmp-3 siRNA or ghrelin treatment compared with vehicle control. Thus, our study indicates that MMP-3 produced in blood vessel endothelial cells after SCI serves as an endogenous molecule for microglial activation followed by p38MAPK activation and proNGF production, and further indicates that the protective effect of ghrelin on oligodendrocytes cell death may be at least partly mediated by the inhibition of MMP-3-induced microglial activation after SCI.
Asunto(s)
Muerte Celular/fisiología , Células Endoteliales/metabolismo , Endotelio Vascular/metabolismo , Metaloproteinasa 3 de la Matriz/metabolismo , Microglía/metabolismo , Oligodendroglía/metabolismo , Traumatismos de la Médula Espinal/metabolismo , Animales , Muerte Celular/efectos de los fármacos , Línea Celular , Células Endoteliales/efectos de los fármacos , Células Endoteliales/patología , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/patología , Ghrelina/farmacología , Ratones , Microglía/efectos de los fármacos , Microglía/patología , Factores de Crecimiento Nervioso/metabolismo , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Neuronas/patología , Oligodendroglía/efectos de los fármacos , Oligodendroglía/patología , Traumatismos de la Médula Espinal/patología , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
Blood-spinal cord barrier (BSCB) disruption and progressive hemorrhage after spinal cord injury (SCI) lead to secondary injury and the subsequent apoptosis and/or necrosis of neuron and glia, causing permanent neurological deficits. In this study, we examined the effect of 17ß-estradiol (E2) on BSCB breakdown and hemorrhage as well as subsequent inflammation after SCI. After a moderate contusion injury at the 9th thoracic segment of spinal cord, E2 (300 µg/kg) was administered by iv injection immediately after SCI, and the same dose of E2 was then administered 6 and 24 hours after injury. Our data show that E2 attenuated BSCB permeability and hemorrhage and reduced the infiltration of neutrophils and macorphages after SCI. Consistent with this finding, the expression of inflammatory mediators was significantly reduced by E2. Furthermore, E2 treatment significantly inhibited the expression of sulfonylurea receptor 1 and transient receptor potential melastatin 4 after injury, which are known to mediate hemorrhage at an early stage after SCI. Moreover, the expression and activation of matrix metalloprotease-9 after injury, which is known to disrupt BSCB, and the degradation of tight junction proteins, such as zona occludens-1 and occludin, were significantly inhibited by E2 treatment. Furthermore, the protective effects of E2 on BSCB disruption and functional improvement were abolished by an estrogen receptor antagonist, ICI 182780 (3 mg/kg). Thus, our study provides evidence that the neuroprotective effect of E2 after SCI is, in part, mediated by inhibiting BSCB disruption and hemorrhage through the down-regulation of sulfonylurea receptor 1/transient receptor potential melastatin 4 and matrix metalloprotease-9, which is dependent on estrogen receptor.
Asunto(s)
Barrera Hematoencefálica/efectos de los fármacos , Permeabilidad Capilar/efectos de los fármacos , Estradiol/farmacología , Hemorragia/metabolismo , Metaloproteinasa 9 de la Matriz/efectos de los fármacos , Traumatismos de la Médula Espinal/metabolismo , Receptores de Sulfonilureas/efectos de los fármacos , Canales Catiónicos TRPM/efectos de los fármacos , Animales , Apoptosis , Barrera Hematoencefálica/metabolismo , Regulación hacia Abajo/efectos de los fármacos , Estradiol/análogos & derivados , Antagonistas del Receptor de Estrógeno/farmacología , Fulvestrant , Masculino , Metaloproteinasa 9 de la Matriz/metabolismo , Ratas , Ratas Sprague-Dawley , Receptores de Sulfonilureas/metabolismo , Canales Catiónicos TRPM/metabolismoRESUMEN
Blood spinal cord barrier (BSCB) disruption after spinal cord injury (SCI) leads to secondary injury and results in apoptosis of neurons and glia, leading to permanent neurological deficits. Here, we examined the effect of ghrelin on BSCB breakdown and hemorrhage after SCI. After moderate weight-drop contusion injury at T9 spinal cord, ghrelin (80µg/kg) was administered via intraperitoneal injection immediately after SCI and then the same dose of ghrelin was treated every 6h for 1d. Our data showed that ghrelin treatment significantly inhibited the expression and activation of matrix metalloprotease-9 (MMP-9) at 1d after SCI. The increases of sulfonylurea receptor 1 (SUR1) and transient receptor potential melastatin 4 (TrpM4) expressions at 1h and 8h after SCI respectively were also alleviated by ghrelin treatment. In addition, both BSCB breakdown and hemorrhage at 1d after injury were significantly attenuated by ghrelin. In parallel, the infiltration of blood cells such as neutrophils and macrophages was inhibited by ghrelin treatment at 1d and 5d after SCI respectively. We also found that ghrelin receptor, growth hormone secretagogue receptor-1a (GHS-R1a), was expressed in the blood vessel of normal spinal tissue. Furthermore, the inhibitory effects of ghrelin on hemorrhage and BSCB disruption at 1d after SCI were blocked by GHS-R1a antagonist, [D-Lys-3]-GHRP-6 (3mg/kg). Thus, these results indicate that the neuroprotective effect by ghrelin after SCI is mediated in part by blocking BSCB disruption and hemorrhage through the down-regulation of SUR1/TrpM4 and MMP-9, which is dependent on GHS-R1a.
Asunto(s)
Ghrelina/farmacología , Metaloproteinasa 9 de la Matriz/genética , Traumatismos de la Médula Espinal/genética , Médula Espinal/efectos de los fármacos , Receptores de Sulfonilureas/genética , Canales Catiónicos TRPM/genética , Animales , Western Blotting , Permeabilidad Capilar/efectos de los fármacos , Expresión Génica/efectos de los fármacos , Ghrelina/administración & dosificación , Hemorragia/prevención & control , Inmunohistoquímica , Mediadores de Inflamación/metabolismo , Inyecciones Intraperitoneales , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Masculino , Metaloproteinasa 9 de la Matriz/metabolismo , Neutrófilos/efectos de los fármacos , Neutrófilos/metabolismo , Oligopéptidos/farmacología , Ratas Sprague-Dawley , Receptores de Ghrelina/antagonistas & inhibidores , Receptores de Ghrelina/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Médula Espinal/irrigación sanguínea , Traumatismos de la Médula Espinal/metabolismo , Traumatismos de la Médula Espinal/fisiopatología , Receptores de Sulfonilureas/metabolismo , Canales Catiónicos TRPM/metabolismo , Factores de TiempoRESUMEN
Occupational hand eczema is frequent in hospital workers, especially in nurses. A comprehensive understanding regarding hand eczema is essential for establishing proper prevention and treatment strategies. The purpose of this study was to identify the risk factors for hand eczema in hospital nursing staffs. A self-administered questionnaire study was performed on hospital nursing staffs at a single general hospital in Korea. In addition, 70 patients with hand eczema underwent patch testing. Five hundred and twenty-five of 700 invited nurses completed the study (response rate, 75.0%). The overall frequency of symptom-based hand eczema was 75.6%, and self-reported hand eczema was 31.0%. Risk factors for hand eczema were young age, history of atopic dermatitis, frequent hand washing (>20 times/day) and long duration of glove wearing (>5 min). Hand eczema was less frequent among frequent hand moisturizer users (>3-4 times/day). Positive patch test reactions were observed in 61.4%. Frequent allergens were nickel sulfate (35.7%), cobalt chloride (28.6%) and thiomersal (21.4%). Among various antibiotics, ciprofloxacin (11.4%), trimethoprim/sulfamethoxazole (11.4%) and gentamicin (7.1%) were revealed as common allergens, in order of frequency. Hand eczema is quite common among hospital nursing staffs. Proper preventive programs and educations are demanded.
Asunto(s)
Dermatitis por Contacto/epidemiología , Eccema/epidemiología , Dermatosis de la Mano/epidemiología , Enfermería/estadística & datos numéricos , Enfermedades Profesionales/epidemiología , Adulto , Alérgenos/efectos adversos , Alérgenos/análisis , Dermatitis por Contacto/inmunología , Eccema/inmunología , Femenino , Dermatosis de la Mano/inmunología , Humanos , Masculino , Persona de Mediana Edad , Enfermedades Profesionales/inmunología , Pruebas del Parche , Prevalencia , República de Corea/epidemiología , Factores de Riesgo , Adulto JovenRESUMEN
As many new cosmetic products are introduced into the market, attention must be given to contact dermatitis, which is commonly caused by cosmetics. We investigate the prevalence of preservative allergy in 584 patients with suspected cosmetic contact dermatitis at 11 different hospitals. From January 2010 to March 2011, 584 patients at 11 hospital dermatology departments presented with cosmetic contact dermatitis symptoms. These patients were patch-tested for preservative allergens. An irritancy patch test performed on 30 control subjects using allergens of various concentrations showed high irritancy rates. Preservative hypersensitivity was detected in 41.1% of patients. Allergens with the highest positive test rates were benzalkonium chloride (12.1%), thimerosal (9.9%) and methylchloroisothiazolinone/methylisothiazolinone (MCI/MI) (5.5%). Benzalkonium chloride and chlorphenesin had the highest irritancy rate based on an irritancy patch test performed using various concentrations. Seven of 30 normal subjects had a positive irritant patch reading with 0.1% benzalkonium chloride and eight of 30 normal subjects had a positive irritant patch reading at 4 days with 0.5% chlorphenesin in petrolatum. Although benzalkonium chloride was highly positive for skin reactions in our study, most reactions were probably irritation. MCI/MI and thimerosal showed highly positive allergy reactions in our study. The optimum concentration of chlorphenesin to avoid skin reactions is less than 0.5%.