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1.
J Microbiol Biotechnol ; 34(6): 1348-1355, 2024 Jun 28.
Artículo en Inglés | MEDLINE | ID: mdl-38755008

RESUMEN

The eukaryotic translation initiation factor eIF5B is a bacterial IF2 ortholog that plays an important role in ribosome joining and stabilization of the initiator tRNA on the AUG start codon during the initiation of translation. We identified the fluorophenyl oxazole derivative 2,2-dibromo-1-(2-(4-fluorophenyl)benzo[d]oxazol-5-yl)ethanone quinolinol as an inhibitor of fungal protein synthesis using an in vitro translation assay in a fungal system. Mutants resistant to this compound were isolated in Saccharomyces cerevisiae and were demonstrated to contain amino acid substitutions in eIF5B that conferred the resistance. These results suggest that eIF5B is a target of potential antifungal compound and that mutation of eIF5B can confer resistance. Subsequent identification of 16 other mutants revealed that primary mutations clustered mainly on domain 2 of eIF5B and secondarily mainly on domain 4. Domain 2 has been implicated in the interaction with the small ribosomal subunit during initiation of translation. The tested translation inhibitor could act by weakening the functional contact between eIF5B and the ribosome complex. This data provides the basis for the development of a new family of antifungals.


Asunto(s)
Antifúngicos , Factores Eucarióticos de Iniciación , Saccharomyces cerevisiae , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/efectos de los fármacos , Factores Eucarióticos de Iniciación/metabolismo , Factores Eucarióticos de Iniciación/genética , Antifúngicos/farmacología , Mutación , Biosíntesis de Proteínas/efectos de los fármacos , Farmacorresistencia Fúngica/genética , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/antagonistas & inhibidores , Inhibidores de la Síntesis de la Proteína/farmacología , Sustitución de Aminoácidos
2.
Biochem Biophys Res Commun ; 417(4): 1254-9, 2012 Jan 27.
Artículo en Inglés | MEDLINE | ID: mdl-22227193

RESUMEN

Casuarinin is a naturally occurring tannin that is isolated from the leaves of Hippophae rhamnoides. It has been shown to have anti-oxidant, anti-cancer, anti-viral, and anti-inflammatory activities. The aim of this study was to investigate the possible mechanism by which casuarinin inhibits TNF-α/IFN-γ-induced Th2 chemokines expression in the human keratinocytes cell line HaCaT. We found that casuarinin suppressed TNF-α/IFN-γ-induced expression of TARC and MDC mRNA and protein in HaCaT cells. Casuarinin significantly inhibited TNF-α/IFN-γ-induced activation of NF-κB, STAT1, and p38 MAPK. Furthermore, we observed that p38 MAPK contributes to inhibition of TNF-α/IFN-γ-induced TARC and MDC production by blocking NF-κB and STAT1 activation in HaCaT cells. Taken together, these results suggest that casuarinin may exert anti-inflammatory responses by suppressing TNF-α/IFN-γ-induced expression of TARC and MDC via blockage of p38 MAPK activation and subsequent activation of NF-κB and STAT1. We propose that it could therefore be used as a therapeutic agent against inflammatory skin diseases.


Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Quimiocina CCL17/antagonistas & inhibidores , Quimiocina CCL22/antagonistas & inhibidores , Taninos Hidrolizables/farmacología , FN-kappa B/antagonistas & inhibidores , Factor de Transcripción STAT1/antagonistas & inhibidores , Antiinflamatorios no Esteroideos/uso terapéutico , Línea Celular , Quimiocina CCL17/biosíntesis , Quimiocina CCL22/biosíntesis , Dermatitis/tratamiento farmacológico , Humanos , Taninos Hidrolizables/uso terapéutico , Interferón gamma/farmacología , Queratinocitos/efectos de los fármacos , Queratinocitos/metabolismo , Factor de Necrosis Tumoral alfa/farmacología , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo
3.
Biochem Biophys Res Commun ; 409(4): 780-5, 2011 Jun 17.
Artículo en Inglés | MEDLINE | ID: mdl-21621513

RESUMEN

Hippophae rhamnoides has been extensively used in oriental traditional medicines for treatment of asthma, skin diseases, gastric ulcers, and lung disorders. In this study, we isolated casuarinin from the leaves of H.rhamnoides and examined the effect of casuarinin on the TNF-α-induced ICAM-1 expression in a human keratinocytes cell line HaCaT. Pretreatment with casuarinin inhibited TNF-α-induced protein and mRNA expression of ICAM-1 and subsequent monocyte adhesiveness in HaCaT cells. Casuarinin significantly inhibited TNF-α-induced NF-κB activation. In addition, casuarinin inhibited activation of ERK and p38 MAPK in a dose-dependent manner. Furthermore, pretreatment with casuarinin decreased TNF-α-induced pro-inflammatory mediators, such as IL-1ß, IL-6, IL-8, and MCP-1. These results demonstrated that casuarinin exerts its anti-inflammatory activity by suppressing TNF-α-induced expression of ICAM-1 and pro-inflammatory cytokines/chemokines via blockage of activation of NF-κB and ERK/p38 MAPK and can be used as a therapeutic agent against inflammatory skin diseases.


Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Taninos Hidrolizables/farmacología , Molécula 1 de Adhesión Intercelular/biosíntesis , Queratinocitos/efectos de los fármacos , FN-kappa B/antagonistas & inhibidores , Antiinflamatorios no Esteroideos/química , Línea Celular , Quimiocina CCL2/antagonistas & inhibidores , Dermatitis/tratamiento farmacológico , Dermatitis/inmunología , Hippophae/química , Humanos , Taninos Hidrolizables/aislamiento & purificación , Interleucina-1beta/antagonistas & inhibidores , Interleucina-6/antagonistas & inhibidores , Interleucina-8/antagonistas & inhibidores , Queratinocitos/inmunología , Hojas de la Planta/química , Factor de Necrosis Tumoral alfa/farmacología
4.
Biochem Biophys Res Commun ; 407(3): 535-40, 2011 Apr 15.
Artículo en Inglés | MEDLINE | ID: mdl-21414301

RESUMEN

We previously demonstrated that celastrol, a quinone methide triterpenoid derived from the medicinal plant Tripterygium wilfordii, exerts its anti-inflammatory activity through up-regulation of heme oxygenase-1 (HO-1) expression in the keratinocytes. In this study, we examined the signaling pathways that lead to the up-regulation of HO-1 expression by celastrol. In HaCaT cells, celastrol-induced HO-1 expression was dependent on ROS generation. ERK and p38 MAPK were major MAPK pathways responsible for celastrol-induced HO-1 expression. Celastrol induced Nrf2 activation. Nrf2 knockdown using small interfering RNA (siRNA) inhibited celastrol-induced HO-1 expression. Treatment with celastrol resulted in a marked increase in antioxidant response element (ARE)-driven transcriptional activity, which was dependent on ROS generation and activation of ERK and p38 MAPK. Furthermore, Nrf2 siRNA significantly reversed the inhibitory effect of celastrol on IFN-γ-induced expression of ICAM-1 in the keratinocytes. Taken together, our results indicate that celastrol can activate the ROS-ERK/p38-Nrf2-ARE signaling cascades leading to the up-regulation of HO-1 which is partly responsible for its anti-inflammatory activity in the keratinocytes.


Asunto(s)
Aciltransferasas/metabolismo , Antiinflamatorios no Esteroideos/farmacología , Hemo-Oxigenasa 1/biosíntesis , Factor 2 Relacionado con NF-E2/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Triterpenos/farmacología , Línea Celular , Hemo-Oxigenasa 1/genética , Humanos , Molécula 1 de Adhesión Intercelular/biosíntesis , Interferón gamma/farmacología , Queratinocitos/efectos de los fármacos , Queratinocitos/metabolismo , Triterpenos Pentacíclicos , Transducción de Señal , Regulación hacia Arriba
5.
Biochem Biophys Res Commun ; 398(1): 140-5, 2010 Jul 16.
Artículo en Inglés | MEDLINE | ID: mdl-20599745

RESUMEN

Celastrol, a quinone methide triterpenoid derived from the medicinal plant Tripterygium wilfordii, possesses various biological activities such as anti-oxidant, anti-tumor, and anti-inflammatory activities. In this study, we examined the suppressive effect of celastrol on IFN-gamma-induced expression of ICAM-1 and the molecular mechanism responsible for these activities. We found that celastrol induced mRNA and protein expression of heme oxygenase-1 (HO-1) in the human keratinocyte cell line HaCaT. Treatment of HaCaT cells with tin protoporphyrin IX (SnPP), a specific inhibitor of HO-1, reversed the suppressive effect of celastrol on IFN-gamma-induced protein and mRNA expression of ICAM-1. HO-1 knockdown using small interfering RNA (siRNA) led to reverse inhibition of IFN-gamma-induced up-regulation of ICAM-1 by celastrol. In addition, SnPP reversed suppression of IFN-gamma-induced promoter activity of ICAM-1 by celastrol. Furthermore, blockage of HO-1 activity by SnPP and HO-1 siRNA reversed the inhibitory effect of celastrol on IFN-gamma-induced adhesion of monocytes to keratinocytes. These results suggest that celastrol may exert anti-inflammatory responses by suppressing IFN-gamma-induced expression of ICAM-1 and subsequent monocyte adhesion via expression of HO-1 in the keratinocytes.


Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Hemo-Oxigenasa 1/biosíntesis , Molécula 1 de Adhesión Intercelular/biosíntesis , Triterpenos/farmacología , Adhesión Celular/efectos de los fármacos , Línea Celular , Inhibidores Enzimáticos/farmacología , Hemo-Oxigenasa 1/antagonistas & inhibidores , Hemo-Oxigenasa 1/genética , Humanos , Interferón gamma/farmacología , Metaloporfirinas/farmacología , Monocitos/efectos de los fármacos , Monocitos/inmunología , Triterpenos Pentacíclicos , Protoporfirinas/farmacología
6.
Biochem Biophys Res Commun ; 387(1): 115-20, 2009 Sep 11.
Artículo en Inglés | MEDLINE | ID: mdl-19576177

RESUMEN

Keratinocytes, one of major cell types in the skin, can be induced by TNF-alpha and IFN-gamma to express thymus- and activation-regulated chemokine (TARC/CCL17), which is considered to be a pivotal mediator in the inflammatory responses during the development of inflammatory skin diseases, such as atopic dermatitis (AD). In this study, we examined the effect of 1,2,3,4,6-penta-O-galloyl-beta-d-glucose (PGG), isolated from the barks of Juglans mandshurica, on TNF-alpha/IFN-gamma induced CCL17 expression in the human keratinocyte cell line HaCaT. Pretreatment of HaCaT cells with PGG suppressed TNF-alpha/IFN-gamma-induced protein and mRNA expression of CCL17. PGG significantly inhibited TNF-alpha/IFN-gamma-induced NF-kappaB activation as well as STAT1 activation. Furthermore, pretreatment with PGG resulted in significant reduction in expression of CXCL9, 10, and 11 in the HaCaT cells treated with IFN-gamma. These results suggest that PGG may exert anti-inflammatory responses by suppressing TNF-alpha and/or IFN-gamma-induced activation of NF-kappaB and STAT1 in the keratinocytes and might be a useful tool in therapy of skin inflammatory diseases.


Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Quimiocina CCL17/biosíntesis , Taninos Hidrolizables/farmacología , Queratinocitos/efectos de los fármacos , FN-kappa B/metabolismo , Factor de Transcripción STAT1/metabolismo , Línea Celular , Humanos , Interferón gamma/farmacología , Juglans/química , Queratinocitos/metabolismo , Factor de Necrosis Tumoral alfa/farmacología
7.
J Med Food ; 19(7): 654-62, 2016 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-27331630

RESUMEN

Abnormal expression of pro-inflammatory mediators such as cell adhesion molecules and cytokines has been implicated in various inflammatory skin diseases, including atopic dermatitis. In this study, we investigated the anti-inflammatory activity of Aronia melanocarpa concentrate (AC) and its action mechanisms using in vivo and in vitro skin inflammation models. Topical application of AC on mouse ears significantly suppressed 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced ear edema formation, as judged by measuring ear thickness and weight, and histological analysis. Topical administration of AC also reduced the expression of pro-inflammatory cytokines such as TNF-α, IL-1ß, and IL-6 in TPA-stimulated mouse ears. Pretreatment with AC suppressed TNF-α-induced ICAM-I expression and subsequent monocyte adhesiveness in human keratinocyte cell line HaCaT. In addition, AC significantly decreased intracellular reactive oxygen species (ROS) generation as well as mitogen-activated protein kinase (MAPK) activation in TNF-α-stimulated HaCaT cells. AC and its constituent cyanidin 3-glucoside also attenuated TNF-α-induced IKK activation, IκB degradation, p65 phosphorylation/nuclear translocation, and p65 DNA binding activity in HaCaT cells. Overall, our results indicate that AC exerts anti-inflammatory activities by inhibiting expression of pro-inflammatory mediators in vitro and in vivo possibly through suppression of ROS-MAPK-NF-κB signaling pathways. Therefore, AC may be developed as a therapeutic agent to treat various inflammatory skin diseases.


Asunto(s)
Antiinflamatorios , Edema/tratamiento farmacológico , Frutas/química , Photinia , Extractos Vegetales/administración & dosificación , Acetato de Tetradecanoilforbol , Animales , Adhesión Celular/efectos de los fármacos , Línea Celular , Dermatitis/tratamiento farmacológico , Oído , Edema/inducido químicamente , Expresión Génica/efectos de los fármacos , Humanos , Molécula 1 de Adhesión Intercelular/genética , Queratinocitos , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Ratones , Monocitos , Factor de Necrosis Tumoral alfa/farmacología
8.
Cancer Lett ; 356(2 Pt B): 743-50, 2015 Jan 28.
Artículo en Inglés | MEDLINE | ID: mdl-25449436

RESUMEN

An adenovirus harboring the HSV thymidine kinase (HSVtk) gene under the regulation of a trans-splicing ribozyme that targets telomerase is cytotoxic to cancer cells because it inhibits DNA replication (Ad5mTR). Furthermore, it induces anti-tumor immunity by activating cytotoxic T cells. Because multiple chemotherapeutic agents also activate cytotoxic T-cell immunity during the direct killing process of tumor cells, we herein explored whether low-dose cisplatin could synergize with cytotoxic Ad5mTR to potentiate its therapeutic effect by boosting anti-tumor immunity in a murine HPV16-associated tonsillar carcinoma model. Tumor regression was enhanced when low-dose (1 mg/kg) cisplatin was added to suicide gene therapy using Ad5mTR. Meanwhile, 1 mg/kg cisplatin alone had no tumor-suppressive effects and did not result in any systemic toxicity. Thus, cisplatin along with Ad5mTR improved tumor clearance by increasing the number of E7-specific CD8+ T cells. Specifically, analysis of the tumors and lymph nodes supported improved immune clearance by increasing the number of E7-specific CD8+ T cells inside tumors (40%, P < 0.05) as a result of the combination of suicide gene and cisplatin therapy. These results suggest that a low dose of cisplatin potentiates CD8+ T-cell-mediated anti-tumor immunity, and its addition to the HSVtk-based adenovirus results in additional therapeutic benefits for HPV16-positive head and neck cancer patients.


Asunto(s)
Carcinoma de Células Escamosas/terapia , Cisplatino/administración & dosificación , Infecciones por Papillomavirus/terapia , Simplexvirus/enzimología , Timidina Quinasa/metabolismo , Neoplasias Tonsilares/terapia , Microambiente Tumoral/efectos de los fármacos , Adenoviridae/genética , Animales , Antineoplásicos/administración & dosificación , Antineoplásicos/farmacología , Western Blotting , Carcinoma de Células Escamosas/inmunología , Carcinoma de Células Escamosas/virología , Ciclo Celular , Proliferación Celular , Cisplatino/farmacología , Terapia Combinada , Relación Dosis-Respuesta a Droga , Papillomavirus Humano 16/patogenicidad , Humanos , Inmunidad Celular/efectos de los fármacos , Inmunidad Celular/inmunología , Técnicas para Inmunoenzimas , Masculino , Ratones , Ratones Endogámicos C57BL , Infecciones por Papillomavirus/inmunología , Infecciones por Papillomavirus/patología , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Linfocitos T Citotóxicos/efectos de los fármacos , Linfocitos T Citotóxicos/inmunología , Timidina Quinasa/genética , Neoplasias Tonsilares/inmunología , Neoplasias Tonsilares/virología , Células Tumorales Cultivadas
9.
Head Neck ; 37(3): 327-35, 2015 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-24700450

RESUMEN

BACKGROUND: The purpose of this study was to characterize the expression of CD200, a membrane protein that functions in immune evasion, to examine its correlations with cancer stem cell (CSC)-like features and analyze its response to chemotherapy and radiation in human papillomavirus (HPV)-positive (+) and negative (-) head and neck squamous cell carcinomas (HNSCCs). METHODS: CD200 expression was analyzed in several HNSCC cell lines. CD200 was overexpressed in HPV(+) murine tonsil epithelial cells, its effects on Shh and Bmi-1 were examined in vitro, and tumor growth and response to chemoradiation were analyzed in vitro and in vivo. RESULTS: CD200 was diversely expressed and consistently associated with expression of Bmi-1 and Shh. Overexpression of CD200 induced Bmi-1 and Shh. Tumors grew similarly between C57BL/6 and Rag1(-/-) C57BL/6 mice. CD200 expression enhanced the resistance to chemoradiation only in vivo. CONCLUSION: CD200 was related to CSC features and modulates response to chemoradiation in vivo. Attenuating this might be a potential therapeutic strategy.


Asunto(s)
Antígenos CD/genética , Carcinoma de Células Escamosas/terapia , Proliferación Celular/genética , Quimioradioterapia/métodos , Neoplasias de Cabeza y Cuello/terapia , Animales , Antígenos CD/inmunología , Carcinoma de Células Escamosas/patología , Línea Celular Tumoral , Citometría de Flujo , Regulación Neoplásica de la Expresión Génica , Neoplasias de Cabeza y Cuello/patología , Humanos , Immunoblotting , Ratones , Ratones Endogámicos C57BL , Neoplasias Experimentales , Células Madre Neoplásicas/efectos de los fármacos , Células Madre Neoplásicas/efectos de la radiación , Carcinoma de Células Escamosas de Cabeza y Cuello , Transfección , Resultado del Tratamiento
10.
Mol Cells ; 33(4): 335-41, 2012 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-22450687

RESUMEN

Defensins, a family of antimicrobial peptides, are one of the first lines of host defense. Human beta-defensins (hBD) such as hBD-2 and -3 have anti-HIV activity. Previous studies have shown that HIV-1 virion can induce the expression of hBD, although the exact components of HIV-1 virion that are responsible for hBD expression have not yet been elucidated. In this study, we examined the effect of HIV-1 Tat on the expression of hBD in B cells. Stimulation of B cells with HIV-1 Tat protein significantly increased the mRNA and protein levels of hBD-2. HIV-1 Tat also induced the activation of a reporter gene for hBD-2 in a dose-dependent manner in B cells. Pretreatment of B cells with a JNK inhibitor suppressed HIV-1 Tat-induced hBD-2 expression. Pretreatment of B cells with AP-1 inhibitors or NF-κB inhibitors led to a decrease in HIV-1 Tat-induced protein and mRNA expression of hBD-2. Taken together, our results indicate that HIV-1 Tat can up-regulate the expression of hBD-2 via JNK-NF-κB/AP-1-dependent pathways in human B cells.


Asunto(s)
Linfocitos B/metabolismo , FN-kappa B , Factor de Transcripción AP-1 , beta-Defensinas , Productos del Gen tat del Virus de la Inmunodeficiencia Humana/genética , Linfocitos B/inmunología , Línea Celular , Dactinomicina/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/inmunología , VIH-1/inmunología , VIH-1/metabolismo , Humanos , MAP Quinasa Quinasa 1/antagonistas & inhibidores , MAP Quinasa Quinasa 1/metabolismo , MAP Quinasa Quinasa 4/antagonistas & inhibidores , MAP Quinasa Quinasa 4/metabolismo , FN-kappa B/genética , FN-kappa B/metabolismo , Factor de Transcripción AP-1/genética , Factor de Transcripción AP-1/metabolismo , Transfección , beta-Defensinas/genética , beta-Defensinas/inmunología , beta-Defensinas/metabolismo , Productos del Gen tat del Virus de la Inmunodeficiencia Humana/metabolismo
11.
BMB Rep ; 44(7): 462-7, 2011 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-21777517

RESUMEN

Up-regulation of selected matrix metalloproteinases (MMPs) such as MMP-9 contributes to inflammatory processes during the development of various skin diseases, such as atopic dermatitis. In this study, we examined the effect of a cell-permeable superoxide dismutase (Tat-SOD) on TNF-α-induced MMP-9 expression in human keratinocyte cells (HaCaT). When Tat-SOD was added to the culture medium of HaCaT cells, it rapidly entered the cells in dose- and time-dependent manners. Tat-SOD decreased TNF-α-induced reactive oxygen species (ROS) generation. Tat-SOD also inhibited TNF-α-induced NF-κB DNA binding activity. Treatment of HaCaT cells with Tat-SOD significantly inhibited TNF-α-induced mRNA and protein expression of MMP-9, as measured by RT-PCR and Western blot analysis. In addition, Tat-SOD suppressed TNF-α-induced gelatinolytic activity of MMP-9. Taken together, our results indicate that Tat-SOD can suppress TNF-α-induced MMP-9 expression via ROS-NF-κB-dependent mechanisms in keratinocytes, and therefore can be used as an immunomodulatory agent against inflammatory skin diseases related to oxidative stress.


Asunto(s)
Permeabilidad de la Membrana Celular/efectos de los fármacos , Queratinocitos/citología , Queratinocitos/enzimología , Metaloproteinasa 9 de la Matriz/metabolismo , Superóxido Dismutasa/metabolismo , Factor de Necrosis Tumoral alfa/farmacología , Línea Celular , ADN/metabolismo , Humanos , Queratinocitos/efectos de los fármacos , FN-kappa B/metabolismo , Unión Proteica/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , Productos del Gen tat del Virus de la Inmunodeficiencia Humana/metabolismo
12.
Free Radic Biol Med ; 50(5): 576-84, 2011 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-21172429

RESUMEN

Up-regulation of adhesion molecules such as vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) by the HIV-1 transactivator of transcription (Tat) in activated microglia and astrocytes may play a pivotal role during the development of AIDS-related encephalitis and dementia. Previous studies demonstrated that HIV-1 Tat-induced up-regulation of adhesion molecules was mediated by reactive oxygen species (ROS), although the mechanisms underlying HIV-1 Tat-induced ROS generation are unknown. In this study, we examined the possible role of NADPH oxidase in HIV-1 Tat-induced up-regulation of adhesion molecules in astroglioma cell lines. HIV-1 Tat-induced up-regulation of VCAM-1/ICAM-1 and subsequent increased adhesion of monocytes to astrocytes were blocked by a general NADPH oxidase inhibitor, diphenylene iodonium, and a specific inhibitor of NADPH oxidase assembly, 9R3A-gp91ds. Nox2 knockdown using small interfering RNA (siRNA) inhibited HIV-1 Tat-induced up-regulation of adhesion molecules and subsequent increased adhesion of monocytes to astrocytes. Nox2 siRNA blocked HIV-1 Tat-induced ROS production, increase in NADPH oxidase activity, and Rac1 activation. Furthermore, Nox2 siRNA decreased HIV-1 Tat-induced NF-κB activation as well as activation of MAP kinases including ERK, JNK, and p38. These data indicate that Nox2-based NADPH oxidase is responsible for HIV-1 Tat-induced generation of ROS and plays an important role in the up-regulation of adhesion molecules such as VCAM-1/ICAM-1 and subsequent increased adhesion of monocytes to astrocytes and serves as a novel target for HIV-1 Tat-mediated neurological diseases.


Asunto(s)
Astrocitos/metabolismo , VIH-1/metabolismo , Molécula 1 de Adhesión Intercelular/genética , Glicoproteínas de Membrana/fisiología , NADPH Oxidasas/fisiología , Especies Reactivas de Oxígeno/metabolismo , Molécula 1 de Adhesión Celular Vascular/genética , Productos del Gen tat del Virus de la Inmunodeficiencia Humana/metabolismo , Complejo SIDA Demencia/metabolismo , Adhesión Celular/genética , Adhesión Celular/fisiología , Línea Celular Tumoral , Técnicas de Silenciamiento del Gen , VIH-1/genética , Humanos , Molécula 1 de Adhesión Intercelular/fisiología , Glicoproteínas de Membrana/antagonistas & inhibidores , Glicoproteínas de Membrana/genética , Monocitos/inmunología , NADPH Oxidasa 2 , NADPH Oxidasas/antagonistas & inhibidores , NADPH Oxidasas/genética , Compuestos Onio/farmacología , ARN Interferente Pequeño/genética , Regulación hacia Arriba/genética , Regulación hacia Arriba/fisiología , Molécula 1 de Adhesión Celular Vascular/fisiología , Productos del Gen tat del Virus de la Inmunodeficiencia Humana/genética
13.
Mol Cells ; 29(3): 245-50, 2010 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-20108167

RESUMEN

Oxidative stress plays a pivotal role in uncontrolled neuro-inflammation leading to many neurological diseases including Alzheimer's. One of the major antioxidant enzymes known to prevent deleterious effects due to oxidative stress is Cu,Zn-superoxide dismutase (SOD). In this study, we examined the regulatory function of SOD on the LPS-induced signaling pathways leading to NF-kappaB activation, expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), in BV-2 cells using cell-permeable SOD. Treatment of BV-2 cells with cell-permeable SOD led to a decrease in LPS-induced reactive oxygen species (ROS) generation and significantly inhibited protein and mRNA levels of iNOS and COX-2 upregulated by LPS. Production of NO and PGE2 in LPS stimulated BV-2 cells was significantly abrogated by pretreatment with a cell-permeable SOD fusion protein. Furthermore, cell-permeable SOD inhibited LPS-induced NF-kappaB DNA-binding activity and activation of MAP kinases including ERK, JNK, and p38 in BV-2 cells. These data indicate that SOD has a regulatory function for LPS-induced NF-kappaB activation leading to expression of iNOS and COX-2 in BV-2 cells and suggest that cell-permeable SOD is a feasible therapeutic agent for regulation of ROS-related neurological diseases.


Asunto(s)
Ciclooxigenasa 2/biosíntesis , Microglía/enzimología , Óxido Nítrico Sintasa de Tipo II/biosíntesis , Superóxido Dismutasa/fisiología , Animales , Línea Celular/efectos de los fármacos , Línea Celular/enzimología , Permeabilidad de la Membrana Celular , Ciclooxigenasa 2/genética , Dinoprostona/biosíntesis , Regulación hacia Abajo/efectos de los fármacos , Inducción Enzimática/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , VIH-1/genética , Humanos , Lipopolisacáridos/farmacología , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Ratones , Microglía/efectos de los fármacos , FN-kappa B/metabolismo , Óxido Nítrico/biosíntesis , Óxido Nítrico Sintasa de Tipo II/genética , Fragmentos de Péptidos/genética , Fragmentos de Péptidos/fisiología , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Recombinantes de Fusión/fisiología , Superóxido Dismutasa/genética , Productos del Gen tat del Virus de la Inmunodeficiencia Humana/genética , Productos del Gen tat del Virus de la Inmunodeficiencia Humana/fisiología
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