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1.
Mol Neurobiol ; 58(11): 5703-5721, 2021 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-34390469

RESUMEN

Dolutegravir (DTG) is a first-line antiretroviral drug (ARV) used in combination therapy for the treatment of human immunodeficiency virus type-1 (HIV-1) infection. The drug is effective, safe, and well tolerated. Nonetheless, concerns have recently emerged for its usage in pregnant women or those of child-bearing age. Notably, DTG-based ARV regimens have been linked to birth defects seen as a consequence of periconceptional usages. To this end, uncovering an underlying mechanism for DTG-associated adverse fetal development outcomes has gained clinical and basic research interest. We now report that DTG inhibits matrix metalloproteinases (MMPs) activities that could affect fetal neurodevelopment. DTG is a broad-spectrum MMPs inhibitor and binds to Zn++ at the enzyme's catalytic domain. Studies performed in pregnant mice show that DTG readily reaches the fetal central nervous system during gestation and inhibits MMP activity. Postnatal screenings of brain health in mice pups identified neuroinflammation and neuronal impairment. These abnormalities persist as a consequence of in utero DTG exposure. We conclude that DTG inhibition of MMPs activities during gestation has the potential to affect prenatal and postnatal neurodevelopment.


Asunto(s)
Antirretrovirales/toxicidad , Compuestos Heterocíclicos con 3 Anillos/toxicidad , Inhibidores de la Metaloproteinasa de la Matriz/toxicidad , Defectos del Tubo Neural/inducido químicamente , Trastornos del Neurodesarrollo/inducido químicamente , Enfermedades Neuroinflamatorias/inducido químicamente , Oxazinas/toxicidad , Piperazinas/toxicidad , Piridonas/toxicidad , Animales , Antirretrovirales/farmacocinética , Antirretrovirales/farmacología , Encéfalo/embriología , Encéfalo/enzimología , Dominio Catalítico/efectos de los fármacos , Femenino , Perfilación de la Expresión Génica , Compuestos Heterocíclicos con 3 Anillos/farmacocinética , Compuestos Heterocíclicos con 3 Anillos/farmacología , Masculino , Inhibidores de la Metaloproteinasa de la Matriz/farmacocinética , Inhibidores de la Metaloproteinasa de la Matriz/farmacología , Ratones , Ratones Endogámicos C3H , Simulación del Acoplamiento Molecular , Defectos del Tubo Neural/embriología , Neuroimagen , Enfermedades Neuroinflamatorias/embriología , Oxazinas/farmacocinética , Oxazinas/farmacología , Piperazinas/farmacocinética , Piperazinas/farmacología , Placenta/química , Embarazo , Piridonas/farmacocinética , Piridonas/farmacología , Distribución Tisular , Zinc/metabolismo
2.
Reprod Sci ; 28(11): 3085-3093, 2021 11.
Artículo en Inglés | MEDLINE | ID: mdl-34050522

RESUMEN

Benzo(a)pyrene (BaP) is a ubiquitous environmental endocrine-disrupting chemical that is known to have toxic effects on reproduction. However, the underlying mechanisms describing how BaP and its metabolite benzo[a]pyrene-7, 8-diol-9, 10-epoxide (BPDE) induce recurrent pregnancy loss (RPL) are still largely unclear. In this study, we identified a novel long non-coding RNA (lnc-HZ07, NCBI MT936329) that was upregulated in trophoblast cells after exposure to BPDE, and lnc-HZ07 expression was significantly higher in RPL villous tissues than that in control villous tissues. Knockdown of lnc-HZ07 promoted trophoblast cell migration, whereas overexpression of lnc-HZ07 inhibited trophoblast cell migration. Further study showed that lnc-HZ07 inhibited trophoblast migration by downregulating matrix metalloproteinase 2 (MMP2) expression via dephosphorylation of AKT. These results demonstrated a novel regulatory pathway in which BaP downregulated AKT phosphorylation and inhibited MMP2 expression by upregulating lnc-HZ07, suggesting that lnc-HZ07 could be considered as a potential pathological marker of BaP-induced RPL and therapeutic target for this disease.


Asunto(s)
Aborto Habitual/metabolismo , Benzo(a)pireno/toxicidad , Metaloproteinasa 2 de la Matriz/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Trofoblastos/efectos de los fármacos , Aborto Habitual/inducido químicamente , Adulto , Animales , Movimiento Celular/efectos de los fármacos , Movimiento Celular/fisiología , Células Cultivadas , Contaminación Ambiental/efectos adversos , Femenino , Humanos , Inhibidores de la Metaloproteinasa de la Matriz/toxicidad , Ratones , Ratones Endogámicos C57BL , Inhibidores de las Quinasa Fosfoinosítidos-3/toxicidad , Embarazo , Proteínas Proto-Oncogénicas c-akt/antagonistas & inhibidores , ARN Largo no Codificante/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Trofoblastos/metabolismo , Trofoblastos/patología , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/fisiología , Adulto Joven
3.
ChemMedChem ; 16(8): 1257-1267, 2021 04 20.
Artículo en Inglés | MEDLINE | ID: mdl-33506625

RESUMEN

Microbial infections are a significant threat to public health, and resistance is on the rise, so new antibiotics with novel modes of action are urgently needed. The extracellular zinc metalloprotease collagenase H (ColH) from Clostridium histolyticum is a virulence factor that catalyses tissue damage, leading to improved host invasion and colonisation. Besides the major role of ColH in pathogenicity, its extracellular localisation makes it a highly attractive target for the development of new antivirulence agents. Previously, we had found that a highly selective and potent thiol prodrug (with a hydrolytically cleavable thiocarbamate unit) provided efficient ColH inhibition. We now report the synthesis and biological evaluation of a range of zinc-binding group (ZBG) variants of this thiol-derived inhibitor, with the mercapto unit being replaced by other zinc ligands. Among these, an analogue with a phosphonate motif as ZBG showed promising activity against ColH, an improved selectivity profile, and significantly higher stability than the thiol reference compound, thus making it an attractive candidate for future drug development.


Asunto(s)
Acetanilidas/farmacología , Proteínas Bacterianas/antagonistas & inhibidores , Quelantes/farmacología , Inhibidores de la Metaloproteinasa de la Matriz/farmacología , Organofosfonatos/farmacología , Acetanilidas/síntesis química , Acetanilidas/toxicidad , Animales , Bacillus cereus/enzimología , Proteínas Bacterianas/metabolismo , Línea Celular Tumoral , Quelantes/síntesis química , Quelantes/toxicidad , Clostridium histolyticum/enzimología , Colágeno/metabolismo , Colagenasas/metabolismo , Células HEK293 , Humanos , Inhibidores de la Metaloproteinasa de la Matriz/síntesis química , Inhibidores de la Metaloproteinasa de la Matriz/toxicidad , Organofosfonatos/síntesis química , Organofosfonatos/toxicidad , Porcinos , Pez Cebra , Zinc/química
4.
Eur J Pharmacol ; 887: 173431, 2020 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-32758568

RESUMEN

Intraocular pressure (IOP) lowering in glaucomatous eyes is currently achieved mainly by improved aqueous outflow via alternate drainage pathways. However, the focus is now shifting to trabecular meshwork (TM), the site or major pathological changes including increased extracellular matrix (ECM) deposition and reduced matrix metalloproteinases (MMPs) secretion by TM cells. Trans-resveratrol was previously shown to lower IOP and reduce ECM deposition; however, the mechanisms of action remain unclear. Therefore, we determined the effect of trans-resveratrol on MMP-2 and -9 expression by human TM cells (HTMCs) in the presence of dexamethasone and whether it also affects adenosine A1 receptors (A1AR) expression and nuclear factor kappa B (NFkB) activation. We observed that trans-resveratrol, 12.5 µM, increased MMP-2 and -9 protein expression by HTMCs despite exposure to dexamethasone (1.89- and 1.53-fold, respectively; P < 0.001). Further it was observed that trans-resveratrol increases A1AR expression in HTMC in the presence of dexamethasone (1.55-fold; P < 0.01). Trans-resveratrol also increased NFkB activation in the presence of dexamethasone and A1AR antagonist (P < 0.01 versus dexamethasone group). These effects of trans-resveratrol were associated with increased MMP -2 and -9 expression. It could be concluded that trans-resveratrol prevents dexamethasone-induced reduction in MMP-2 and -9 secretion by NFkB activation in HTMCs. This effect of trans-resveratrol is likely to involve increased A1AR expression.


Asunto(s)
Dexametasona/toxicidad , Metaloproteinasas de la Matriz/biosíntesis , FN-kappa B/biosíntesis , Receptor de Adenosina A1/biosíntesis , Resveratrol/farmacología , Malla Trabecular/metabolismo , Antioxidantes/farmacología , Células Cultivadas , Regulación Enzimológica de la Expresión Génica , Humanos , Inhibidores de la Metaloproteinasa de la Matriz/toxicidad , FN-kappa B/antagonistas & inhibidores , Malla Trabecular/efectos de los fármacos
5.
J Korean Med Sci ; 32(4): 666-671, 2017 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-28244295

RESUMEN

We evaluated the safety of matrix metalloproteinase (MMP) inhibitor in experimental glaucoma filtration surgery in an animal model. Fifteen New Zealand white rabbits underwent an experimental trabeculectomy and were randomly allocated into 3 groups according to the adjuvant agent: no treatment group (n = 5), 0.02% mitomycin C (MMC) soaking group (n = 5), and MMP inhibitor (ilomastat) subconjunctival injection group (n = 5). Slit lamp examination with Seidel testing, pachymetry, and specular microscopy was performed preoperatively and postoperatively. The conjunctiva and ciliary body toxicity were evaluated with scores according to the pathologic grading systems. Electron microscopy was used to examine the structural changes in cornea, conjunctiva, and ciliary body. In the ilomastat-treated group, there was no statistically significant change in central corneal thickness preoperatively and at 28 days postoperatively (P = 0.655). There were also no significant changes in specular microscopy findings over the duration of the study in the ilomastat-treated group. The conjunctival toxicity score was 1 in the control group, 1.5 in the ilomastat-treated group, and 2 in the MMC-treated group. When assessing ciliary body toxicity scores, the ilomastat-treated group score was 0.5 and the MMC-treated group score was 1.5. Transmission electron microscopy did not show structural changes in the cornea and ciliary body whereas the structural changes were noticed in MMC group. A single subconjunctival injection of MMP inhibitor during the experimental trabeculectomy showed a less toxic affect in the rabbit cornea, conjunctiva, and ciliary body compared to MMC.


Asunto(s)
Conjuntiva/efectos de los fármacos , Glaucoma/cirugía , Indoles/toxicidad , Inhibidores de la Metaloproteinasa de la Matriz/toxicidad , Animales , Cuerpo Ciliar/efectos de los fármacos , Cuerpo Ciliar/fisiología , Conjuntiva/fisiología , Córnea/efectos de los fármacos , Córnea/fisiología , Ácidos Hidroxámicos , Microscopía Electrónica de Transmisión , Mitomicina/toxicidad , Conejos , Trabeculectomía
6.
J Dent Res ; 94(8): 1085-91, 2015 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-26040283

RESUMEN

Nanoparticles (NPs) are currently the focus of considerable attention for dental applications; however, their biological effects have not been fully elucidated. The long-term, slow release of matrix metalloproteases (MMPs) digests collagen fibrils within resin-dentin bonds. Therefore, MMP inhibitors can prolong the durability of resin-dentin bonds. However, there have been few reports evaluating the combined effect of MMP inhibition and the cytotoxic effects of NPs for dentin bonding. The aim of this study was to evaluate MMP inhibition and cytotoxic responses to gold (AuNPs) and platinum nanoparticles (PtNPs) stabilized by polyvinylpyrrolidone (PVP) in cultured murine macrophages (RAW264) by using MMP inhibition assays, measuring cell viability and inflammatory responses (quantitative reverse transcription polymerase chain reaction [RT-qPCR]), and conducting a micromorphological analysis by fluorescence and transmission electron microscopy. Cultured RAW264 cells were exposed to metal NPs at various concentrations (1, 10, 100, and 400 µg/mL). AuNPs and PtNPs markedly inhibited MMP-8 and MMP-9 activity. Although PtNPs were cytotoxic at high concentrations (100 and 400 µg/mL), no cytotoxic effects were observed for AuNPs at any concentration. Transmission electron microscopy images showed a significant nonrandom intercellular distribution for AuNPs and PtNPs, which were mostly observed to be localized in lysosomes but not in the nucleus. RT-qPCR analysis demonstrated inflammatory responses were not induced in RAW264 cells by AuNPs or PtNPs. The cytotoxicity of nanoparticles might depend on the core metal composition and arise from a "Trojan horse" effect; thus, MMP inhibition could be attributed to the surface charge of PVP, which forms the outer coating of NPs. The negative charge of the surface coating of PVP binds to Zn(2+) from the active center of MMPs by chelate binding and results in MMP inhibition. In summary, AuNPs are attractive NPs that effectively inhibit MMP activity without cytotoxicity or inflammatory responses.


Asunto(s)
Oro/química , Oro/toxicidad , Macrófagos/efectos de los fármacos , Inhibidores de la Metaloproteinasa de la Matriz/química , Inhibidores de la Metaloproteinasa de la Matriz/toxicidad , Nanopartículas/química , Nanopartículas/toxicidad , Animales , Supervivencia Celular/efectos de los fármacos , Humanos , Inmunohistoquímica , Ratones , Microscopía Electrónica de Transmisión , Microscopía Fluorescente , Platino (Metal)/química , Platino (Metal)/toxicidad , Povidona/química , Povidona/toxicidad , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
7.
Biochem Biophys Res Commun ; 457(4): 538-41, 2015 Feb 20.
Artículo en Inglés | MEDLINE | ID: mdl-25600809

RESUMEN

Matrix metalloproteinases (MMPs) are zinc (Zn(2+)) and calcium (Ca(2+)) dependant endopeptidases, capable of degradation of numerous components of the extracellular matrix. Cadmium (Cd(2+)) is a well known environmental contaminant which could impair the activity of MMPs. In this sense, this study was conducted to evaluate if Cd(2+) intake inhibits these endopeptidases activities at the rat prostate and testicles and if it directly inhibits the activity of MMP2 and MMP9 at gelatinolytic assays when present in the incubation buffer. To investigate this hypothesis, Wistar rats (5 weeks old), were given tap water (untreated, n = 9), or 15 ppm CdCl2 diluted in drinking water, during 10 weeks (n = 9) and 20 weeks (n = 9). The animals were euthanized and their ventral prostate, dorsal prostate, and testicles were removed. These tissue samples were processed for protein extraction and subjected to gelatin zymography evaluation. Additionally, we performed an experiment of gelatin zymography in which 5 µM or 2 mM cadmium chloride (CdCl2) was directly dissolved at the incubation buffer, using the prostatic tissue samples from untreated animals that exhibited the highest MMP2 and MMP9 activities in the previous experiment. We have found that CdCl2 intake in the drinking water led to the inhibition of 35% and 30% of MMP2 and MMP9 (p < 0.05) at the ventral prostate and testis, respectively, in Cd(2+) treated animals when compared to controls. Moreover, the activities of the referred enzymes were 80% and 100% inhibited by 5 µM and 2 mM of CdCl2, respectively, even in the presence of 10 mM of CaCl2 within the incubation buffer solution. These important findings demonstrate that environmental cadmium contamination may deregulate the natural balance in the extracellular matrix turnover, through MMPs downregulation, which could contribute to the toxic effects observed in prostatic and testicular tissue after its exposure.


Asunto(s)
Cadmio/toxicidad , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Inhibidores de la Metaloproteinasa de la Matriz/toxicidad , Próstata/enzimología , Testículo/enzimología , Contaminantes Químicos del Agua/toxicidad , Animales , Masculino , Próstata/efectos de los fármacos , Ratas Wistar , Testículo/efectos de los fármacos
8.
J Periodontol ; 85(6): 868-75, 2014 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-24215203

RESUMEN

BACKGROUND: Recent studies have shown that epigallocatechin-3-gallate (EGCG), a major constituent of green tea extract, exhibits effects of anti-inflammation and antioxidation on periodontal inflammation. The present in vitro study examines the effect of EGCG on Porphyromonas gingivalis (Pg) lipopolysaccharide (LPS)-enhanced expression of interleukin (IL)-6 and matrix metalloproteinase (MMP)-1, as well as the activation of nuclear factor-kappa B (NF-κB). Furthermore, the role of IL-6 on LPS-enhanced MMP-1 production is evaluated using human gingival fibroblasts (HGFs). METHODS: HGFs were primary cultured from human gingiva specimens. The cytotoxicities of EGCG and LPS were tested by cell viability tests. The cellular mRNA expression of IL-6 was determined by reverse-transcription polymerase chain reaction, and the protein expression of MMP-1 and IL-6 was examined by enzyme-linked immunosorbent assay. The cytosol expression and nuclear translocation of NF-κB was evaluated by immunocytochemistry followed by confocal laser scanning microscopy. RESULTS: Pg LPS significantly increased MMP-1 production in HGFs, whereas adding EGCG significantly attenuated this enhanced production of MMP-1. LPS treatment also increased the mRNA and protein expression of IL-6 and stimulated NF-κB activation in HGFs. However, the addition of EGCG significantly attenuated the IL-6 expression and NF-κB activation. Supplemental addition of IL-6 significantly enhanced cellular MMP-1 production, whereas anti-IL-6 antibody inhibited LPS-enhanced MMP-1 production. CONCLUSION: EGCG could attenuate Pg LPS-enhanced production of MMP-1 in HGFs, whereas this attenuation might be due to the inhibition of IL-6 by EGCG.


Asunto(s)
Antioxidantes/farmacología , Catequina/análogos & derivados , Fibroblastos/efectos de los fármacos , Encía/efectos de los fármacos , Interleucina-6/antagonistas & inhibidores , Lipopolisacáridos/farmacología , Metaloproteinasa 1 de la Matriz/efectos de los fármacos , Inhibidores de la Metaloproteinasa de la Matriz/farmacología , Porphyromonas gingivalis/efectos de los fármacos , Adulto , Antioxidantes/toxicidad , Catequina/farmacología , Catequina/toxicidad , Técnicas de Cultivo de Célula , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Femenino , Encía/citología , Humanos , Interleucina-6/análisis , Masculino , Metaloproteinasa 1 de la Matriz/análisis , Inhibidores de la Metaloproteinasa de la Matriz/toxicidad , Microscopía Confocal , FN-kappa B/efectos de los fármacos , Adulto Joven
9.
Bioorg Med Chem ; 21(21): 6456-65, 2013 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-24071448

RESUMEN

The complexity of matrix metalloproteinase inhibitors (MMPIs) design derives from the difficulty in carefully addressing their inhibitory activity towards the MMP isoforms involved in many pathological conditions. In particular, specific metalloproteinases, such as MMP-2 and MMP-9, are key regulators of the 'vicious cycle' occurring between tumor metastases growth and bone remodeling. In an attempt to devise new approaches to selective inhibitor derivatives, we describe novel bisphosphonate bone seeking MMP inhibitors (BP-MMPIs), capable to be selectively targeted and to overcome undesired side effects of broad spectrum MMPIs. In vitro activity (IC50 values) for each inhibitor was determined against MMP-2, -8, -9 and -14, because of their relevant role in skeletal development and renewal. The results show that BP-MMPIs reached IC50 values of enzymatic inhibition in the low micromolar range. Computational studies, used to rationalize some trends in the observed inhibitory profiles, suggest a possible differential binding mode in MMP-2 that explains the selective inhibition of this isoform. In addition, survival assay was conducted on J774 cell line, a well known model system used to evaluate the structure-activity relationship of BPs for inhibiting bone resorption. The resulting data, confirming the specific activity of BP-MMPIs, and their additional proved propensity to bind hydroxyapatite powder in vitro, suggest a potential use of BP-MMPIs in skeletal malignancies.


Asunto(s)
Difosfonatos/química , Inhibidores de la Metaloproteinasa de la Matriz/química , Metaloproteinasas de la Matriz/química , Animales , Sitios de Unión , Línea Celular , Supervivencia Celular/efectos de los fármacos , Difosfonatos/metabolismo , Difosfonatos/toxicidad , Durapatita/química , Durapatita/metabolismo , Células Hep G2 , Humanos , Metaloproteinasa 14 de la Matriz/química , Metaloproteinasa 14 de la Matriz/metabolismo , Metaloproteinasa 2 de la Matriz/química , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 8 de la Matriz/química , Metaloproteinasa 8 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/química , Metaloproteinasa 9 de la Matriz/metabolismo , Inhibidores de la Metaloproteinasa de la Matriz/metabolismo , Inhibidores de la Metaloproteinasa de la Matriz/toxicidad , Metaloproteinasas de la Matriz/metabolismo , Ratones , Simulación del Acoplamiento Molecular , Unión Proteica , Estructura Terciaria de Proteína , Relación Estructura-Actividad
10.
J Med Chem ; 56(11): 4357-73, 2013 Jun 13.
Artículo en Inglés | MEDLINE | ID: mdl-23631440

RESUMEN

New series of pyrrolidine mercaptosulfide, 2-mercaptocyclopentane arylsulfonamide, and 3-mercapto-4-arylsulfonamidopyrrolidine matrix metalloproteinase inhibitors (MMPIs) were designed, synthesized, and evaluated. Exhibiting unique properties over other MMPIs (e.g., hydroxamates), these newly reported compounds are capable of modulating activities of several MMPs in the low nanomolar range, including MMP-2 (~2 to 50 nM), MMP-13 (~2 to 50 nM), and MMP-14 (~4 to 60 nM). Additionally these compounds are selective to intermediate- and deep-pocket MMPs but not shallow-pocketed MMPs (e.g., MMP-1, ~850 to >50,000 nM; MMP-7, ~4000 to >25,000 nM). Our previous work with the mercaptosulfide functionality attached to both cyclopentane and pyrrolidine frameworks demonstrated that the cis-(3S,4R)-stereochemistry was optimal for all of the MMPs tested. However, in our newest compounds an interesting shift of preference to the trans form of the mercaptosulfonamides was observed with increased oxidative stability and biological compatibility. We also report several kinetic and biological characteristics showing that these compounds may be used to probe the mechanistic activities of MMPs in disease.


Asunto(s)
Ciclopentanos/síntesis química , Inhibidores de la Metaloproteinasa de la Matriz/síntesis química , Pirrolidinas/síntesis química , Compuestos de Sulfhidrilo/síntesis química , Sulfonamidas/síntesis química , Células Cultivadas , Ciclopentanos/química , Ciclopentanos/toxicidad , Estabilidad de Medicamentos , Humanos , Cinética , Inhibidores de la Metaloproteinasa de la Matriz/química , Inhibidores de la Metaloproteinasa de la Matriz/toxicidad , Metaloproteinasas de la Matriz/química , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/efectos de los fármacos , Pirrolidinas/química , Pirrolidinas/toxicidad , Estereoisomerismo , Relación Estructura-Actividad , Compuestos de Sulfhidrilo/química , Compuestos de Sulfhidrilo/toxicidad , Sulfonamidas/química , Sulfonamidas/toxicidad
11.
Biochem Pharmacol ; 85(12): 1770-82, 2013 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-23603294

RESUMEN

Infliximab, a monoclonal antibody directed against human tumor necrosis factor-alpha (TNF-α), effectively treats anterior uveitis, which can accompany Behçet's disease. Here, we investigated the underlying mechanism of this action. We examined human, non-pigmented ciliary epithelial cells (HNPCECs), which make up the blood-aqueous barrier (BAB) in the uvea. We measured the expression levels of matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs in the presence or absence of TNF-α using quantitative, real-time polymerase chain reaction and enzyme-linked immunosorbent assays. The expression of MMP-1, MMP-3, and MMP-9 increased in the presence of TNF-α, and the addition of infliximab reversed the increase. The TNF-α effects were more attenuated when infliximab was added before than when it was added after TNF-α exposure. Gelatin zymography demonstrated that the protease activity of these MMPs was also increased in the presence of TNF-α and attenuated with infliximab. Immunostaining showed that MMP-1, MMP-3, and MMP-9 degraded claudin-1 and occludin in HNPCECs and in non-pigmented ciliary epithelial cells of the swine ciliary body. In a monolayer of HNPCECs, we found that permeability was significantly increased with MMP treatment. Thus, TNF-α increased levels of MMPs in cells that form the BAB, and MMPs degraded components of the tight junctions in the BAB, which increased permeability through the cellular barrier. Furthermore, infliximab effectively attenuated the TNF-α-induced increases in MMP expression in cells that make up the BAB. These findings might suggest a basis for the clinical prevention of anterior uveitis.


Asunto(s)
Anticuerpos Monoclonales/farmacología , Cuerpo Ciliar/metabolismo , Claudina-1/antagonistas & inhibidores , Metaloproteinasas de la Matriz/metabolismo , Ocludina/antagonistas & inhibidores , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Factor de Necrosis Tumoral alfa/toxicidad , Animales , Células Cultivadas , Cuerpo Ciliar/efectos de los fármacos , Claudina-1/metabolismo , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/inmunología , Inducción Enzimática/efectos de los fármacos , Inducción Enzimática/fisiología , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Humanos , Infliximab , Inhibidores de la Metaloproteinasa de la Matriz/metabolismo , Inhibidores de la Metaloproteinasa de la Matriz/toxicidad , Metaloproteinasas de la Matriz/biosíntesis , Ocludina/metabolismo , Porcinos
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