Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 1.658
Filtrar
1.
J Enzyme Inhib Med Chem ; 36(1): 1829-1838, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34353202

RESUMO

As the largest organ in human body, skin acts as a physicochemical barrier, offering protection against harmful environmental stressors, such as chemicals, pathogens, temperature and radiation. Nonetheless, skins prominence goes further, with a significant psychosocial role in an increasingly ageing population. Prompted by consumers' concern regarding skincare, cosmetic industry has been developing new formulas capable of lessening the most visible signs of ageing, including reduction in skin density and elasticity, wrinkling and hyperpigmentation. Allied to skincare is the rising importance set on natural products, sustainably obtained from less environmental impacting methods. Cyanobacteria and microalgae are adding importance in this field, given their ability to biosynthesize secondary metabolites with anti-ageing potential. In this review, we present an overview on the potential of cyanobacteria and microalgae compounds to overcome skin-ageing, essentially by exploring their effects on the metalloproteinases collagenase, elastase, gelatinase and hyaluronidase, and in other enzymes involved in the pigmentation process.


Assuntos
Antioxidantes/química , Misturas Complexas/química , Cianobactérias/química , Matriz Extracelular/metabolismo , Hiperpigmentação/tratamento farmacológico , Microalgas/química , Envelhecimento da Pele/efeitos dos fármacos , Antioxidantes/farmacologia , Produtos Biológicos/química , Colagenases/metabolismo , Misturas Complexas/farmacologia , Gelatinases/metabolismo , Humanos , Hialuronoglucosaminidase/metabolismo , Metaloproteases/metabolismo , Elastase Pancreática/metabolismo , Pele
3.
J Enzyme Inhib Med Chem ; 36(1): 1665-1678, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34309457

RESUMO

Oleanolic acid (OA) is a natural cosmeceutical compound with various skin beneficial activities including inhibitory effect on hyaluronidase but the anti-hyaluronidase activity and mechanisms of action of its synthetic analogues remain unclear. Herein, a series of OA derivatives were synthesised and evaluated for their inhibitory effects on hyaluronidase. Compared to OA, an induction of fluorinated (6c) and chlorinated (6g) indole moieties led to enhanced anti-hyaluronidase activity (IC50 = 80.3 vs. 9.97 and 9.57 µg/mL, respectively). Furthermore, spectroscopic and computational studies revealed that 6c and 6g can bind to hyaluronidase protein and alter its secondary structure leading to reduced enzyme activity. In addition, OA indole derivatives showed feasible skin permeability in a slightly acidic environment (pH = 6.5) and 6c exerted skin protective effect by reducing cellular reactive oxygen species in human skin keratinocytes. Findings from the current study support that OA indole derivatives are potential cosmeceuticals with anti-hyaluronidase activity.


Assuntos
Inibidores Enzimáticos/farmacologia , Hialuronoglucosaminidase/antagonistas & inibidores , Indóis/farmacologia , Ácido Oleanólico/farmacologia , Pele/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Humanos , Hialuronoglucosaminidase/metabolismo , Indóis/síntese química , Indóis/química , Simulação de Acoplamento Molecular , Estrutura Molecular , Ácido Oleanólico/síntese química , Ácido Oleanólico/química , Permeabilidade/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Pele/metabolismo , Relação Estrutura-Atividade
4.
JCI Insight ; 6(17)2021 09 08.
Artigo em Inglês | MEDLINE | ID: mdl-34314391

RESUMO

Vascular injury has emerged as a complication contributing to morbidity in coronavirus disease 2019 (COVID-19). The glycosaminoglycan hyaluronan (HA) is a major component of the glycocalyx, a protective layer of glycoconjugates that lines the vascular lumen and regulates key endothelial cell functions. During critical illness, as in the case of sepsis, enzymes degrade the glycocalyx, releasing fragments with pathologic activities into circulation and thereby exacerbating disease. Here, we analyzed levels of circulating glycosaminoglycans in 46 patients with COVID-19 ranging from moderate to severe clinical severity and measured activities of corresponding degradative enzymes. This report provides evidence that the glycocalyx becomes significantly damaged in patients with COVID-19 and corresponds with severity of disease. Circulating HA fragments and hyaluronidase, 2 signatures of glycocalyx injury, strongly associate with sequential organ failure assessment scores and with increased inflammatory cytokine levels in patients with COVID-19. Pulmonary microvascular endothelial cells exposed to COVID-19 milieu show dysregulated HA biosynthesis and degradation, leading to production of pathological HA fragments that are released into circulation. Finally, we show that HA fragments present at high levels in COVID-19 patient plasma can directly induce endothelial barrier dysfunction in a ROCK- and CD44-dependent manner, indicating a role for HA in the vascular pathology of COVID-19.


Assuntos
COVID-19/metabolismo , Endotélio Vascular/metabolismo , Ácido Hialurônico/metabolismo , Idoso , COVID-19/sangue , COVID-19/patologia , Citocinas/sangue , Endotélio Vascular/patologia , Feminino , Glicocálix/metabolismo , Glicocálix/patologia , Humanos , Receptores de Hialuronatos/metabolismo , Ácido Hialurônico/sangue , Hialuronoglucosaminidase/sangue , Hialuronoglucosaminidase/metabolismo , Masculino , Pessoa de Meia-Idade , Quinases Associadas a rho/metabolismo
5.
Cell Death Dis ; 12(7): 622, 2021 06 16.
Artigo em Inglês | MEDLINE | ID: mdl-34135314

RESUMO

Accumulated evidence shows that OGT-mediated O-GlcNAcylation plays an important role in response to DNA damage repair. However, it is unclear if the "eraser" O-GlcNAcase (OGA) participates in this cellular process. Here, we examined the molecular mechanisms and biological functions of OGA in DNA damage repair, and found that OGA was recruited to the sites of DNA damage and mediated deglycosylation following DNA damage. The recruitment of OGA to DNA lesions is mediated by O-GlcNAcylation events. Moreover, we have dissected OGA using deletion mutants and found that C-terminal truncated OGA including the pseudo HAT domain was required for the recruitment of OGA to DNA lesions. Using unbiased protein affinity purification, we found that the pseudo HAT domain was associated with DNA repair factors including NONO and the Ku70/80 complex. Following DNA damage, both NONO and the Ku70/80 complex were O-GlcNAcylated by OGT. The pseudo HAT domain was required to recognize NONO and the Ku70/80 complex for their deglycosylation. Suppression of the deglycosylation prolonged the retention of NONO at DNA lesions and delayed NONO degradation on the chromatin, which impaired non-homologus end joining (NHEJ). Collectively, our study reveals that OGA-mediated deglycosylation plays a key role in DNA damage repair.


Assuntos
Antígenos de Neoplasias/metabolismo , Núcleo Celular/enzimologia , Dano ao DNA , Reparo do DNA por Junção de Extremidades , Proteínas de Ligação a DNA/metabolismo , Histona Acetiltransferases/metabolismo , Hialuronoglucosaminidase/metabolismo , Autoantígeno Ku/metabolismo , Processamento de Proteína Pós-Traducional , Proteínas de Ligação a RNA/metabolismo , Antígenos de Neoplasias/genética , Linhagem Celular Tumoral , Núcleo Celular/genética , Núcleo Celular/efeitos da radiação , Proliferação de Células , Proteínas de Ligação a DNA/genética , Glicosilação , Células HEK293 , Histona Acetiltransferases/genética , Humanos , Hialuronoglucosaminidase/genética , Autoantígeno Ku/genética , Domínios Proteicos , Proteínas de Ligação a RNA/genética , Especificidade por Substrato
6.
Eur J Med Chem ; 223: 113649, 2021 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-34186233

RESUMO

O-GlcNAcylation is a dynamic post-translational modification mediated by O-linked ß-N-acetylglucosamine transferase (OGT) and O-GlcNAc hydrolase (OGA), that adds or removes a single ß-N-acetylglucosamine (GlcNAc) moiety to or from serine/threonine residues of nucleocytosolic and mitochondrial proteins, respectively. The perturbed homeostasis of O-GlcNAc cycling results in several pathological conditions. Human OGA is a promising therapeutic target in diseases where aberrantly low levels of O-GlcNAc are experienced, such as tauopathy in Alzheimer's disease. A new class of potent OGA inhibitors, 2-acetamido-2-deoxy-d-glucono-1,5-lactone (thio)semicarbazones, have been identified. Eight inhibitors were designed and synthesized in five steps starting from d-glucosamine and with 15-55% overall yields. A heterologous OGA expression protocol with strain selection and isolation has been optimized that resulted in stable, active and full length human OGA (hOGA) isomorph. Thermal denaturation kinetics of hOGA revealed environmental factors affecting hOGA stability. From kinetics experiments, the synthesized compounds proved to be efficient competitive inhibitors of hOGA with Ki-s in the range of ∼30-250 nM and moderate selectivity with respect to lysosomal ß-hexosaminidases. In silico studies consisting of Prime protein-ligand refinements, QM/MM optimizations and QM/MM-PBSA binding free energy calculations revealed the factors governing the observed potencies, and led to design of the most potent analogue 2-acetamido-2-deoxy-d-glucono-1,5-lactone 4-(2-naphthyl)-semicarbazone 6g (Ki = 36 nM). The protocol employed has applications in future structure based inhibitor design targeting OGA.


Assuntos
Antígenos de Neoplasias/metabolismo , Inibidores Enzimáticos/química , Histona Acetiltransferases/metabolismo , Hialuronoglucosaminidase/metabolismo , Lactonas/química , Semicarbazonas/química , Antígenos de Neoplasias/genética , Sítios de Ligação , Inibidores Enzimáticos/metabolismo , Histona Acetiltransferases/antagonistas & inibidores , Histona Acetiltransferases/genética , Humanos , Hialuronoglucosaminidase/antagonistas & inibidores , Hialuronoglucosaminidase/genética , Cinética , Ligantes , Simulação de Dinâmica Molecular , Ligação Proteica , Estabilidade Proteica , Teoria Quântica , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação , Semicarbazonas/metabolismo , Relação Estrutura-Atividade
7.
ACS Appl Mater Interfaces ; 13(23): 26790-26799, 2021 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-34061496

RESUMO

For melanoma with high lethality and metastasis rate, traditional therapy has limited effects; local photothermal therapy (PTT) synergetic with immune therapy for cancer treatment can perhaps improve the situation. However, because of the natural existence of the tumor matrix barrier, the penetration depth of drugs and immune cells often dampens the efficacy of cancer treatment. Herein, we report an innovative synergetic PTT and immune therapy through dissolving microneedles for the codelivery of the hyaluronidase-modified semiconductor polymer nanoparticles containing poly(cyclopentadithiophene-alt-benzothiadiazole) and immune adjuvant polyinosinic-polycytidylic acid (PIC). Benefiting from the dissolution of an extracellular matrix of hyaluronidase, the semiconductor polymer nanoparticles and PIC penetrate the tumor deeply, under synergetic therapy with PTT, activating the immune cells and enhancing the T-cell immune response for inhibition of tumor growth and metastasis. This study provides a promising platform for effective melanoma treatment and a novel strategy to overcome the stromal barrier.


Assuntos
Matriz Extracelular/metabolismo , Hialuronoglucosaminidase/metabolismo , Melanoma Experimental/terapia , Nanopartículas/administração & dosagem , Terapia Fototérmica , Polímeros/química , Linfócitos T/imunologia , Animais , Feminino , Melanoma Experimental/imunologia , Melanoma Experimental/patologia , Camundongos , Camundongos Endogâmicos C57BL , Nanopartículas/química , Agulhas
8.
J Plast Reconstr Aesthet Surg ; 74(7): 1610-1614, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34024740

RESUMO

Hyaluronidase is a family of enzymes that degrade hyaluronic acid (HA). It is found to increase vascular permeability and temporarily disrupt the extracellular matrix, promoting diffusion of substances through tissues. Alongside its applications in ophthalmology, obstetrics and gynaecology, musculoskeletal medicine, radiology and drug and fluid administration, hyaluronidase has a number of roles in the field of plastic surgery. The popularity of HA fillers in recent years has led to an increase in the usage of hyaluronidase in the treatment of filler-related complications. The purpose of this article is to review the current and future uses of hyaluronidase within the field of plastic surgery. Hyaluronidase is used as an adjunct to local anaesthetics in skin infiltration, skin graft harvesting, tumescent analgesia, managing complications of dermal fillers, treatment of extravasation injury, prevention and management of oedema, treatment of ganglion and management of scars. However, it has some limitations. Hyaluronidase is known to interact with a number of common medications. Several case reports also highlight the risk of allergic reaction to the substance. Although rare and usually mild, hyaluronidase has the potential to cause anaphylaxis. Other adverse effects include bruising and swelling. Overall, hyaluronidase appears to be a very safe, cheap and effective medication for a variety of uses in the field of plastic surgery and beyond.


Assuntos
Adjuvantes Anestésicos , Anestesia Local/métodos , Cicatriz/tratamento farmacológico , Preenchedores Dérmicos/efeitos adversos , Ácido Hialurônico/efeitos adversos , Hialuronoglucosaminidase/uso terapêutico , Procedimentos Cirúrgicos Reconstrutivos , Preenchedores Dérmicos/metabolismo , Hipersensibilidade a Drogas/etiologia , Humanos , Ácido Hialurônico/metabolismo , Hialuronoglucosaminidase/efeitos adversos , Hialuronoglucosaminidase/metabolismo
9.
Anal Methods ; 13(17): 2030-2036, 2021 05 07.
Artigo em Inglês | MEDLINE | ID: mdl-33955975

RESUMO

Aggregation induced emission (AIE) dots have gained broad attention in fluorescence bioimaging and biosensors in virtue of their distinctive optical properties of splendid biocompatibility, high brightness and good photostability. However, the application of AIE dots in sensing and imaging of enzymes in cells remains at an early stage and needs to be further explored. In this report, we proposed a novel AIE-dot-based nanoprobe for hyaluronidase (HAase) detection using a simple electrostatic self-assembly of AIE dots with gold nanoparticles functionalized using hyaluronic acid (HA-AuNPs), named HA-AuNPs@AIEDs. The fluorescence of AIE dots can be obviously quenched by HA-AuNPs via fluorescence resonance energy transfer (FRET). HAase could degrade HA into small pieces and thus induce disassembly of AuNPs and AIEDs, accompanied by fluorescence recovery of AIEDs. The as-prepared nanoprobe exhibited high sensitivity, excellent selectivity, wide response range and desirable anti-interference for quantitative sensing of HAase in vitro. The detection limit was down to 0.0072 U mL-1. Moreover, the nanoprobe displayed good biocompatibility and excellent photostability, and thus offered a practicable "turn-on" strategy for specific, high-contrast fluorescence imaging of HAase in live tumor cells. The AIE-based nanoprobe may provide a novel universal platform for recognition and imaging of HAase in tumors, and may be beneficial for related biological research.


Assuntos
Hialuronoglucosaminidase , Nanopartículas Metálicas , Transferência Ressonante de Energia de Fluorescência , Ouro , Ácido Hialurônico , Hialuronoglucosaminidase/metabolismo
10.
Enzyme Microb Technol ; 146: 109759, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33812558

RESUMO

Hyaluronidases are low expressed toxins of brown spider venoms, but, as highly active molecules, they present an important role as spreading factors. By degrading extracellular matrix components, these enzymes favor the diffusion of toxins in the affected tissue and at systemic level. Here, a novel isoform of hyaluronidase of Loxosceles intermedia Mello-Leitão (1934) venom was cloned, expressed in a baculovirus-insect cell expression system and fully active purified. This recombinant enzyme, named LiHyal2 (Loxosceles intermedia Hyaluronidase isoform 2), shares high identity with hyaluronidases of other spiders and scorpions. The catalytic and sugar binding amino acid residues are conserved in LiHyal2, human, and honeybee venom hyaluronidases and the molecular model of LiHyal2 shares major similarities with their crystal structures, including the active site. LiHyal2 was expressed as a 45 kDa protein and degraded hyaluronic acid (HA) and chondroitin sulphate as demonstrated by HA zymography and agarose gel electrophoresis. Lectin blot analysis revealed that LiHyal2 is post-translationally modified by the addition of high mannose N-linked carbohydrates. In vivo experiments showed that LiHyal2 potentialize dermonecrosis and edema induced by a recombinant phospholipase-D (PLD) of L. intermedia venom, as well as enhance the increase in capillary permeability triggered by this PLD, indicating that these toxins act synergistically during envenomation. Altogether, these results introduce a novel approach to express spider recombinant toxins, contribute to the elucidation of brown spider venom mechanisms and add to the development of a more specific treatment of envenomation victims.


Assuntos
Hialuronoglucosaminidase , Fosfolipase D , Animais , Baculoviridae/genética , Baculoviridae/metabolismo , Domínio Catalítico , Humanos , Hialuronoglucosaminidase/genética , Hialuronoglucosaminidase/metabolismo , Insetos/metabolismo , Diester Fosfórico Hidrolases
11.
Carbohydr Polym ; 264: 118015, 2021 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-33910717

RESUMO

Owing to its outstanding water-retention ability, viscoelasticity, biocompatibility and non-immunogenicity, Hyaluronic acid (HA), a natural linear polymer alternating linked by d-glucuronic acid and N-acetylglucosamine, has been widely employed in cosmetic, medical and clinical applications. With the development of synthetic biology and bioprocessing optimization, HA production via microbial fermentation is an economical and sustainable alternative over traditional animal extraction methods. Indeed, recently Streptococci and other recombinant systems for HA synthesis has received increasing interests due to its technical advantages. This review summarizes the production of HA by microorganisms and demonstrates its synthesis mechanism, focusing on the current status in various production systems, as well as common synthetic biology strategies include driving more carbon flux into HA biosynthesis and regulating the molecular weight (MW), and finally discusses the major challenges and prospects.


Assuntos
Ácido Hialurônico/biossíntese , Ácido Hialurônico/química , Animais , Fermentação , Humanos , Hialuronoglucosaminidase/metabolismo , Microbiologia Industrial/métodos , Peso Molecular , Polímeros/química , Streptococcus/crescimento & desenvolvimento , Streptococcus/metabolismo , Biologia Sintética/métodos , Viscosidade
12.
Biomolecules ; 11(3)2021 03 05.
Artigo em Inglês | MEDLINE | ID: mdl-33808040

RESUMO

Non-covalent interactions in supramolecular chemistry provide useful systems to understand biological processes, and self-assembly systems are suitable assets to build-up innovative products for biomedical applications. In this field, polyelectrolyte complexes are interesting, especially when polysaccharides are involved, due to their non-toxicity and bio-absorbability. In this work, we investigated a polyelectrolyte formed by hyaluronic acid (HA), a negatively charged linear polysaccharide, with Chitlac (Ch), a positively charged lactose-modified chitosan. The aim of the study was the investigation of a novel Ch-HA polyelectrolyte complex, to understand the interaction between the two polysaccharides and the stability towards enzymatic activity. By means of gel permeation chromatography-triple detector array (GPC-TDA), nuclear magnetic resonance (NMR), dynamic viscosity, Zeta Potential and scanning electron microscopy (SEM), the polyelectrolyte complex properties were identified and compared to individual polysaccharides. The complex showed monodisperse molecular weight distribution, high viscosity, negative charge, and could be degraded by specific enzymes, such as hyaluronidase and lysozyme. The results suggest a close interaction between the two polysaccharides in the complex, which could be considered a self-assembly system.


Assuntos
Biopolímeros/química , Quitosana/química , Ácido Hialurônico/química , Lactose/química , Animais , Humanos , Hialuronoglucosaminidase/química , Hialuronoglucosaminidase/metabolismo , Espectroscopia de Ressonância Magnética , Microscopia Eletrônica de Varredura , Reologia
13.
Int J Mol Sci ; 22(6)2021 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-33809827

RESUMO

With the burgeoning interest in hyaluronic acid (HA) in recent years, hyaluronidases (HYALs) have come to light for their role in regulating catabolism of HA and its molecular weight (MW) distribution in various tissues. Of the six hyaluronidase-like gene sequences in the human genome, HYALs 1 and 2 are of particular significance because they are the primary hyaluronidases active in human somatic tissue. Perhaps more importantly, for the sake of this review, they cleave anti-inflammatory and anti-fibrotic high-molecular-weight HA into pro-inflammatory and pro-fibrotic oligosaccharides. With this, HYALs regulate HA degradation and thus the development and progression of various diseases. Given the dearth of literature focusing specifically on HYALs in the past decade, this review seeks to expound their role in human diseases of the skin, heart, kidneys, and more. The review will delve into the molecular mechanisms and pathways of HYALs and discuss current and potential future therapeutic benefits of HYALs as a clinical treatment.


Assuntos
Suscetibilidade a Doenças , Hialuronoglucosaminidase/genética , Hialuronoglucosaminidase/metabolismo , Animais , Gerenciamento Clínico , Desenvolvimento de Medicamentos , Regulação da Expressão Gênica , Humanos , Ácido Hialurônico/metabolismo , Hialuronoglucosaminidase/química , Hialuronoglucosaminidase/uso terapêutico , Família Multigênica , Especificidade de Órgãos/genética
14.
Nat Chem Biol ; 17(5): 593-600, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33686291

RESUMO

O-linked N-acetylglucosamine (O-GlcNAc) is an essential and dynamic post-translational modification that is presented on thousands of nucleocytoplasmic proteins. Interrogating the role of O-GlcNAc on a single target protein is crucial, yet challenging to perform in cells. Herein, we developed a nanobody-fused split O-GlcNAcase (OGA) as an O-GlcNAc eraser for selective deglycosylation of a target protein in cells. After systematic cellular optimization, we identified a split OGA with reduced inherent deglycosidase activity that selectively removed O-GlcNAc from the desired target protein when directed by a nanobody. We demonstrate the generality of the nanobody-fused split OGA using four nanobodies against five target proteins and use the system to study the impact of O-GlcNAc on the transcription factors c-Jun and c-Fos. The nanobody-directed O-GlcNAc eraser provides a new strategy for the functional evaluation and engineering of O-GlcNAc via the selective removal of O-GlcNAc from individual proteins directly in cells.


Assuntos
Antígenos de Neoplasias/metabolismo , Histona Acetiltransferases/metabolismo , Hialuronoglucosaminidase/metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Glicoproteínas de Membrana/metabolismo , Complexo de Proteínas Formadoras de Poros Nucleares/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Anticorpos de Domínio Único/química , Fator de Transcrição Sp1/metabolismo , Fatores de Transcrição/metabolismo , Antígenos de Neoplasias/química , Antígenos de Neoplasias/genética , Bioensaio , Domínio Catalítico , Sistemas de Liberação de Medicamentos/métodos , Expressão Gênica , Glicosilação , Células HEK293 , Histona Acetiltransferases/química , Histona Acetiltransferases/genética , Humanos , Hialuronoglucosaminidase/química , Hialuronoglucosaminidase/genética , Hidrólise , Proteínas Quinases JNK Ativadas por Mitógeno/genética , Glicoproteínas de Membrana/genética , Complexo de Proteínas Formadoras de Poros Nucleares/genética , Plasmídeos/química , Plasmídeos/metabolismo , Ligação Proteica , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Anticorpos de Domínio Único/metabolismo , Fator de Transcrição Sp1/genética , Fatores de Transcrição/genética , Transfecção/métodos
15.
J Biol Chem ; 296: 100481, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33647313

RESUMO

The extracellular matrix (ECM) plays an important role in maintaining tissue homeostasis and poses a significant physical barrier to in vivo cell migration. Accordingly, as a means of enhancing tissue invasion, tumor cells use matrix metalloproteinases to degrade ECM proteins. However, the in vivo ECM is comprised not only of proteins but also of a variety of nonprotein components. Hyaluronan (HA), one of the most abundant nonprotein components of the interstitial ECM, forms a gel-like antiadhesive barrier that is impenetrable to particulate matter and cells. Mechanisms by which tumor cells penetrate the HA barrier have not been addressed. Here, we demonstrate that transmembrane protein 2 (TMEM2), the only known transmembrane hyaluronidase, is the predominant mediator of contact-dependent HA degradation and subsequent integrin-mediated cell-substrate adhesion. We show that a variety of tumor cells are able to eliminate substrate-bound HA in a tightly localized pattern corresponding to the distribution of focal adhesions (FAs) and stress fibers. This FA-targeted HA degradation is mediated by TMEM2, which itself is localized at site of FAs. TMEM2 depletion inhibits the ability of tumor cells to attach and migrate in an HA-rich environment. Importantly, TMEM2 directly binds at least two integrins via interaction between extracellular domains. Our findings demonstrate a critical role for TMEM2-mediated HA degradation in the adhesion and migration of cells on HA-rich ECM substrates and provide novel insight into the early phase of FA formation.


Assuntos
Ácido Hialurônico/metabolismo , Proteínas de Membrana/metabolismo , Animais , Adesão Celular/fisiologia , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Movimento Celular/fisiologia , Matriz Extracelular/metabolismo , Adesões Focais/metabolismo , Adesões Focais/fisiologia , Humanos , Receptores de Hialuronatos/metabolismo , Ácido Hialurônico/fisiologia , Hialuronoglucosaminidase/metabolismo , Integrinas/metabolismo , Proteínas de Membrana/fisiologia , Camundongos
16.
Int J Mol Sci ; 22(4)2021 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-33669634

RESUMO

Little is known about the effects on hyaluronan (HA) metabolism of UVA radiation. This study demonstrates that the secretion of HA by human dermal fibroblasts (HDFs) is downregulated by UVA, accompanied by the down- and upregulation of mRNA and protein levels of the HA-synthesizing enzyme (HAS2) and the HA-degrading protein, HYaluronan Binding protein Involved in HA Depolymerization(HYBID), respectively. Signaling analysis revealed that the exposure distinctly elicits activation of the p38/MSK1/CREB/c-Fos/AP-1 axis, the JNK/c-Jun axis, and the p38/ATF-2 axis, but downregulates the phosphorylation of NF-kB and JAK/STAT3. A signal inhibition study demonstrated that the inhibition of p38 significantly abrogates the UVA-accentuated mRNA level of HYBID. Furthermore, the inhibition of STAT3 significantly downregulates the level of HAS2 mRNA in non-UVA exposed HDFs. Analysis using siRNAs demonstrated that transfection of ATF-2 siRNA but not c-Fos siRNA abrogates the increased protein level of HYBID in UVA-exposed HDFs. An inhibitor of protein tyrosine phosphatase but not of protein serine/threonine phosphatase restored the diminished phosphorylation level of STAT3 at Tyr 705, accompanied by a significant abolishing effect on the decreased mRNA expression level of HAS2. Silencing with a protein tyrosine phosphatase PTP-Meg2 siRNA revealed that it abrogates the decreased phosphorylation of STAT3 at Tyr 705 in UVA-exposed HDFs. These findings suggest that the UVA-induced decrease in HA secretion by HDFs is attributable to the down- and upregulation of HAS2 and HYBID expression, respectively, changes that are mainly ascribed to the inactivated signaling of the STAT3 axis due to the activated tyrosine protein phosphatase PTP-Meg2 and the activated signaling of the p38/ATF2 axis, respectively.


Assuntos
Regulação para Baixo/efeitos da radiação , Fibroblastos/efeitos da radiação , Hialuronan Sintases/metabolismo , Ácido Hialurônico/metabolismo , Hialuronoglucosaminidase/metabolismo , Transdução de Sinais/efeitos da radiação , Raios Ultravioleta , Regulação para Cima/efeitos da radiação , Fator 2 Ativador da Transcrição/metabolismo , Morte Celular/efeitos dos fármacos , Morte Celular/efeitos da radiação , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Derme/citologia , Regulação para Baixo/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Humanos , Janus Quinase 2/metabolismo , Masculino , Modelos Biológicos , Peso Molecular , Fosforilação/efeitos dos fármacos , Fosforilação/efeitos da radiação , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fator de Transcrição STAT3/metabolismo , Regulação para Cima/efeitos dos fármacos , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
17.
J Enzyme Inhib Med Chem ; 36(1): 785-789, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33733989

RESUMO

Acute Coronary Syndromes (ACS) with plaque erosion display dysregulated hyaluronan metabolism, with increased hyaluronidase-2 (HYAL2) expression. However, the expression and the role of this enzyme on platelets has never been explored. We evaluated the platelet's HYAL2 (pltHYAL2) levels on I) stable angina (SA) and II) ACS patients, furtherly sub-grouped in Intact-Fibrous-Cap (IFC) and Ruptured-Fibrous-Cap (RFC), according to Optical Coherence Tomography. We assessed the HYAL2 role through an in vitro model setting of co-cultured monocytes and platelets, before and after treatment with low-molecular-weight hyaluronic acid (HA) as pro-inflammatory stimulus and with or without HYAL2-antibody to inhibit HYAL2 activity. ACS patients exhibit higher pltHYAL2 levels comparing to SA, with the higher expression for IFC group. The addition of HYAL2-antibody significantly reduced the percentage of monocyte-platelet binding, suggesting that pltHYAL2 enrichment at the site of the culprit lesion is a key mediator in the systemic thrombo-inflammatory status of ACS presenting with plaque erosion.


Assuntos
Síndrome Coronariana Aguda/metabolismo , Moléculas de Adesão Celular/metabolismo , Hialuronoglucosaminidase/metabolismo , Monócitos/metabolismo , Síndrome Coronariana Aguda/tratamento farmacológico , Síndrome Coronariana Aguda/patologia , Moléculas de Adesão Celular/antagonistas & inibidores , Moléculas de Adesão Celular/sangue , Técnicas de Cocultura , Proteínas Ligadas por GPI/antagonistas & inibidores , Proteínas Ligadas por GPI/sangue , Proteínas Ligadas por GPI/metabolismo , Humanos , Ácido Hialurônico/metabolismo , Ácido Hialurônico/farmacologia , Hialuronoglucosaminidase/antagonistas & inibidores , Hialuronoglucosaminidase/sangue , Monócitos/efeitos dos fármacos , Agregação Plaquetária/efeitos dos fármacos
18.
Am J Physiol Heart Circ Physiol ; 320(4): H1724-H1737, 2021 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-33710913

RESUMO

The surface of vascular endothelial cells (ECs) is covered by a protective negatively charged layer known as the endothelial glycocalyx. Herein, we hypothesized its transport barrier and mechanosensory role in transmural water flux and low-density lipoprotein (LDL) transport in an isolated rat abdominal aorta perfused under 85 mmHg and 20 dyn/cm2 ex vivo. The endothelial glycocalyx was digested by hyaluronidase (HAase) from bovine tests. Water infiltration velocity (Vw) was measured by a graduated pipette. LDL coverage and mean maximum infiltration distance (MMID) in the vessel wall were quantified by confocal laser scanning microscopy. EC apoptosis was determined by the terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) technique, and leaky junction rates were evaluated by electron microscopy. The results showed that a 42% degradation of the endothelial glycocalyx by HAase treatment increased Vw, LDL coverage, and MMID. Shear stress increased Vw, which cannot be inhibited by HAase treatment. Four hour-shear application increased about fourfolds of LDL coverage, whereas exerted no significant effects on its MMID, EC apoptosis, and the leaky junctions. On the contrary, 24-h shear exposure has no significant effects on LDL coverage, whereas increased 2.74-folds of MMID and about 53% of EC apoptotic rates that could be inhibited by HAase treatment. These results suggest endothelial glycocalyx acts as a transport barrier by decreasing water and LDL transport, as well as a mechanosensor of shear to regulate EC apoptosis, thus affecting leaky junctions and regulating LDL transport into the vessel wall.NEW & NOTEWORTHY A 42% degradation of the endothelial glycocalyx by hyaluronidase of the isolated rat abdominal aorta facilitated water and LDL transport across the vessel wall, suggesting endothelial glycocalyx as a transport barrier. A 24-h shear exposure increased LDL mean maximum infiltration distance, and enhanced EC apoptosis, which could be both inhibited by hyaluronidase treatment, suggesting endothelial glycocalyx may also act as a mechanosensor of shear to regulate EC apoptosis, thus affecting leaky junctions and regulating LDL transport.


Assuntos
Aorta Abdominal/metabolismo , Células Endoteliais/metabolismo , Glicocálix/metabolismo , Lipoproteínas LDL/metabolismo , Água/metabolismo , Animais , Aorta Abdominal/ultraestrutura , Apoptose , Transporte Biológico , Células Endoteliais/ultraestrutura , Glicocálix/ultraestrutura , Hialuronoglucosaminidase/metabolismo , Técnicas In Vitro , Masculino , Mecanotransdução Celular , Permeabilidade , Ratos Sprague-Dawley , Fluxo Sanguíneo Regional , Estresse Mecânico
19.
Nat Rev Cancer ; 21(4): 217-238, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33589810

RESUMO

The extracellular matrix is a fundamental, core component of all tissues and organs, and is essential for the existence of multicellular organisms. From the earliest stages of organism development until death, it regulates and fine-tunes every cellular process in the body. In cancer, the extracellular matrix is altered at the biochemical, biomechanical, architectural and topographical levels, and recent years have seen an exponential increase in the study and recognition of the importance of the matrix in solid tumours. Coupled with the advancement of new technologies to study various elements of the matrix and cell-matrix interactions, we are also beginning to see the deployment of matrix-centric, stromal targeting cancer therapies. This Review touches on many of the facets of matrix biology in solid cancers, including breast, pancreatic and lung cancer, with the aim of highlighting some of the emerging interactions of the matrix and influences that the matrix has on tumour onset, progression and metastatic dissemination, before summarizing the ongoing work in the field aimed at developing therapies to co-target the matrix in cancer and cancer metastasis.


Assuntos
Proteínas da Matriz Extracelular/metabolismo , Matriz Extracelular/metabolismo , Metaloproteinases da Matriz/metabolismo , Neoplasias/metabolismo , Inibidores Teciduais de Metaloproteinases/metabolismo , Proteínas ADAM/metabolismo , Proteínas ADAMTS/metabolismo , Proteína Morfogenética Óssea 1/metabolismo , Catepsinas/metabolismo , Movimento Celular , Colágeno/metabolismo , Cistatinas/metabolismo , Elastina/metabolismo , Matriz Extracelular/enzimologia , Matriz Extracelular/patologia , Fibrilinas/metabolismo , Glucuronidase/metabolismo , Glicoproteínas/metabolismo , Humanos , Hialuronoglucosaminidase/metabolismo , Invasividade Neoplásica , Metástase Neoplásica , Neoplasias/enzimologia , Neoplasias/patologia , Processamento de Proteína Pós-Traducional , Proteoglicanas/metabolismo , Serpinas/metabolismo , Metaloproteases Semelhantes a Toloide/metabolismo , Microambiente Tumoral
20.
J Ethnopharmacol ; 270: 113779, 2021 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-33418032

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Caralluma species are traditional edible herbs used in folkloric medicine as antidiabetic, antioxidant, antipyretic, antirheumatic, anti-inflammatory and anthelmintic agents. C. quadrangula was selected in this study to document the traditional use of the genus as anti-rheumatic treatment and the possible mechanisms of action. AIM OF THE STUDY: The higher mortality rates and shorter survival among the patients suffering from rheumatoid arthritis (RA) led to the increased interest on searching for new treatments for RA. Russelioside B (RB), a major pregnane glycoside found in C. quadrangula, was evaluated as a new anti-rheumatic agent. MATERIALS AND METHODS: The n-butanol fraction of C. quadrangula was chromatographed on a silica gel column to isolate RB. The adjuvant-induced arthritis (AIA) model was established in rats by intradermal injection of complete Freund's adjuvant (CFA) to evaluate its anti-arthritic effect. Ibuprofen was used as a reference drug. Forty rats were randomly divided into 5 groups (n = 8): normal (NOR); CFA model (CFA); ibuprofen, 5 mg/kg; RB, 25 mg/kg and RB, 50 mg/kg. The treatments were initiated from day 16 when AIA model was established and continued up to day 40. Serum diagnostic rheumatoid markers, inflammatory cytokines, oxidative stress biomarkers, cartilage and bone degeneration enzymes were assessed. RESULTS: RB at 50 mg/kg b. wt., showed significant decreases in the activities of hyaluronidase and ß-glucouronidase enzymes as well significant decreases in the levels of proinflammatory cytokines as nuclear factor-kappa-B (NF-κB), tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1ß (IL-1ß) compared to the CFA group; 11.04 ± 0.61 pg/mg protein, 4.35 ± 0.25 pg/mg protein, 3.32 ± 0.13 pg/mg protein & 2.75 ± 0.14 pg/mg protein for RB, 50 mg/kg b. wt. group vs. 25.33 ± 2.13 pg/mg protein, 25.65 ± 2.1 pg/mg protein, 22.20 ± 1.34 pg/mg protein & 13.27 ± 1.40 pg/mg protein for the arthritic group, respectively. The total antioxidant capacity (TAC) was significantly restored to normal values in RB, 50 mg/kg treated rats (4.01 ± 0.09 nmol/mL vs. 3.71 ± 0.27 nmol/mL) and the levels of myeloperoxidase (MPO) reduced by 10-folds of the CFA arthritic group. Bone histomorphometry revealed that RB treatment significantly attenuated the CFA-induced bone loss in a dose-dependent manner. CONCLUSIONS: These findings suggested that the anti-arthritic effect of RB was mediated through the reduction of the rheumatoid markers, anti-inflammatory and antioxidant action, inhibition of cartilage and bone degenerative enzymes as well as attenuation of bone loss and osteoclastogenesis.


Assuntos
Anti-Inflamatórios/uso terapêutico , Antirreumáticos/uso terapêutico , Artrite Experimental/tratamento farmacológico , Glicosídeos/uso terapêutico , Pregnanos/uso terapêutico , 1-Butanol/química , Animais , Articulação do Tornozelo/efeitos dos fármacos , Articulação do Tornozelo/patologia , Anticorpos Anti-Proteína Citrulinada/sangue , Anticorpos Anti-Proteína Citrulinada/efeitos dos fármacos , Anti-Inflamatórios/isolamento & purificação , Antirreumáticos/isolamento & purificação , Apocynaceae/química , Artrite Experimental/metabolismo , Contagem de Células Sanguíneas , Peso Corporal/efeitos dos fármacos , Osso Esponjoso/efeitos dos fármacos , Osso Esponjoso/metabolismo , Proteínas de Transporte/sangue , Proteínas de Transporte/efeitos dos fármacos , Citocinas/sangue , Citocinas/efeitos dos fármacos , Edema/tratamento farmacológico , Adjuvante de Freund/toxicidade , Glucuronidase/efeitos dos fármacos , Glucuronidase/metabolismo , Glicosídeos/isolamento & purificação , Hialuronoglucosaminidase/efeitos dos fármacos , Hialuronoglucosaminidase/metabolismo , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Masculino , Medicina Tradicional , Estresse Oxidativo/efeitos dos fármacos , Peroxidase/metabolismo , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/uso terapêutico , Pregnanos/isolamento & purificação , Ratos Wistar , Fator Reumatoide/sangue , Fator Reumatoide/efeitos dos fármacos
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...