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1.
Nat Commun ; 10(1): 2951, 2019 07 04.
Artigo em Inglês | MEDLINE | ID: mdl-31273212

RESUMO

Epithelial-mesenchymal transition (EMT) is an essential process both in physiological and pathological contexts. Intriguingly, EMT is often associated with tissue invagination during development; however, the impact of EMT on tissue remodeling remain unexplored. Here, we show that at the initiation of the EMT process, cells produce an apico-basal force, orthogonal to the surface of the epithelium, that constitutes an important driving force for tissue invagination in Drosophila. When EMT is ectopically induced, cells starting their delamination generate an orthogonal force and induce ectopic folding. Similarly, during mesoderm invagination, cells undergoing EMT generate an apico-basal force through the formation of apico-basal structures of myosin II. Using both laser microdissection and in silico physical modelling, we show that mesoderm invagination does not proceed if apico-basal forces are impaired, indicating that they constitute driving forces in the folding process. Altogether, these data reveal the mechanical impact of EMT on morphogenesis.


Assuntos
Drosophila melanogaster/embriologia , Transição Epitelial-Mesenquimal , Epitélio/embriologia , Morfogênese , Animais , Polaridade Celular , Simulação por Computador , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/citologia , Drosophila melanogaster/metabolismo , Embrião não Mamífero/citologia , Embrião não Mamífero/metabolismo , Epitélio/metabolismo , Mesoderma/citologia , Mesoderma/embriologia , Mesoderma/metabolismo , Modelos Moleculares , Miosina Tipo II/metabolismo
2.
Cancer Sci ; 110(8): 2658-2666, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31199029

RESUMO

Although direct adhesion of cancer cells to the mesothelial cell layer is considered to be a key step for peritoneal invasion of ovarian cancer cell masses (OCM), we recently identified a different strategy for the peritoneal invasion of OCM. In 6 out of 20 cases of ovarian carcinoma, extraperitoneal growth of the OCM was observed along with the neovascularization of feeding vessels, which connect the intraperitoneal host stroma and extraperitoneal lesions through the intact mesothelial cell layer. As an early step, the OCMs anchor in the extraperitoneal fibrin networks and then induce the migration of CD34-positive and vascular endothelial growth factor A (VEGF-A)-positive endothelial cells, constructing extraperitoneal vascular networks around the OCM. During the extraperitoneal growth of OCM, podoplanin-positive and α smooth muscle actin (αSMA)-positive cancer-associated fibroblasts (CAF) appears. In more advanced lesions, the boundary line of mesothelial cells disappears around the insertion areas of feeding vessels and then extraperitoneal and intraperitoneal stroma are integrated, enabling the OCM to invade the host stroma, being associated with CAF. In addition, tissue factors (TF) are strongly detected around these peritoneal implantation sites and their levels in ascites were higher than that in blood. These findings demonstrate the presence of neovascularization around fibrin net-anchored OCMs on the outer side of the intact peritoneal surface, suggesting a novel strategy for peritoneal invasion of ovarian cancer and TF-targeted intraperitoneal anti-cancer treatment. We observed and propose a novel strategy for peritoneal implantation of ovarian cancer. The strategy includes the preinvasive growth of fibrin-anchored cancer cells along with neovascularization on the outer side of the intact peritoneal surface.


Assuntos
Fibrina/metabolismo , Invasividade Neoplásica/patologia , Neovascularização Patológica/patologia , Neoplasias Ovarianas/patologia , Adulto , Idoso , Ascite/metabolismo , Ascite/patologia , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Epitélio/metabolismo , Epitélio/patologia , Feminino , Humanos , Pessoa de Meia-Idade , Neovascularização Patológica/metabolismo , Neoplasias Ovarianas/metabolismo , Neoplasias Peritoneais/metabolismo , Neoplasias Peritoneais/patologia , Peritônio/metabolismo , Peritônio/patologia , Fator A de Crescimento do Endotélio Vascular/metabolismo
3.
Nat Commun ; 10(1): 2406, 2019 06 03.
Artigo em Inglês | MEDLINE | ID: mdl-31160622

RESUMO

Organ-specific colonization suggests that specific cell-cell recognition is essential. Yet, very little is known about this particular interaction. Moreover, tumor cell lodgement requires binding under shear stress, but not static, conditions. Here, we successfully isolate the metastatic populations of cancer stem/tumor-initiating cells (M-CSCs). We show that the M-CSCs tether more and roll slower than the non-metastatic (NM)-CSCs, thus resulting in the preferential binding to the peritoneal mesothelium under ascitic fluid shear stress. Mechanistically, this interaction is mediated by P-selectin expressed by the peritoneal mesothelium. Insulin-like growth factor receptor-1 carrying an uncommon non-sulfated sialyl-Lewisx (sLex) epitope serves as a distinct P-selectin binding determinant. Several glycosyltransferases, particularly α1,3-fucosyltransferase with rate-limiting activity for sLex synthesis, are highly expressed in M-CSCs. Tumor xenografts and clinical samples corroborate the relevance of these findings. These data advance our understanding on the molecular regulation of peritoneal metastasis and support the therapeutic potential of targeting the sLex-P-selectin cascade.


Assuntos
Líquido Ascítico , Carcinoma/secundário , Adesão Celular , Hidrodinâmica , Células-Tronco Neoplásicas/metabolismo , Oligossacarídeos/metabolismo , Neoplasias Ovarianas/patologia , Selectina-P/metabolismo , Neoplasias Peritoneais/secundário , Animais , Carcinoma/metabolismo , Linhagem Celular Tumoral , Epitélio/metabolismo , Feminino , Fucosiltransferases/metabolismo , Células HEK293 , Humanos , Camundongos , Metástase Neoplásica , Transplante de Neoplasias , Neoplasias Ovarianas/metabolismo , Neoplasias Peritoneais/metabolismo , Peritônio/metabolismo , Receptor IGF Tipo 1/metabolismo , Estresse Mecânico
4.
Mol Med Rep ; 19(6): 5464-5472, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31059062

RESUMO

We previously reported that the collapse of ATP production via mitochondrial damage causes ATPase dysfunction, resulting in the onset or progression of lens opacification in cataracts in model rats. In the present study, it was investigated whether the mRNA expression levels of the three subtypes of mitochondrial cytochrome c oxidase (MTCO)1, 2 and 3 and ATP content change with the type and severity of cataracts in human lens. Samples of lens epithelium were collected from Japanese patients during cataract surgery, and the type and severity of the cataracts (grade) were determined according to the WHO classification [cortical (COR), nuclear (NUC), posterior subcapsular (PSC) opacification]. The MTCO1­3 mRNA expression levels in patients with grade­1 COR, NUC and PSC opacification were significantly enhanced compared with those of normal patients. The enhanced MTCO1­3 mRNA levels subsequently decreased in patients with COR, and the MTCO1­3 mRNA levels and ATP levels in patients with grade­3 COR were similar to those in normal patients. However, the mRNA expression levels of MTCO3 in patients with grade 3­NUC opacification and MTCO1­3 in patients with grade­3 PSC opacification, along with the ATP content, were significantly lower than in patients without cataracts. In conclusion, it was revealed that ATP production in lens epithelium is enhanced in early­stage cataracts (grade­1) in Japanese patients with COR, NUC and PSC opacification. In addition, in severe cataracts (grade­3), ATP production and content are strongly decreased in Japanese patients with PSC opacification. ATP depletion in human lens epithelium with PSC opacification may promote lens opacification by ATPase dysfunction.


Assuntos
Catarata/patologia , Complexo IV da Cadeia de Transporte de Elétrons/genética , Cristalino/metabolismo , Mitocôndrias/metabolismo , RNA Mensageiro/metabolismo , Trifosfato de Adenosina/metabolismo , Idoso , Catarata/metabolismo , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Epitélio/metabolismo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Índice de Gravidade de Doença
5.
Methods Mol Biol ; 1966: 27-38, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31041737

RESUMO

Immunohistochemistry using formalin-fixed, paraffin-embedded tissue, chromogen label, and light microscopy has traditionally been used to semiquantify estrogen receptor (ER) to guide diagnosis and management of breast cancer. Quantitation of ER for this purpose currently only assesses levels of the ER-alpha subtype. Considerable variability in results reported has been due to protocol and fixation variability, intraobserver and interobserver variability, and different scoring systems and thresholds for scoring ER positivity. Results can also vary with low expression levels of ER. ER-beta expression is reduced in breast and ovarian cancers and requires quantitation.Herein we describe a novel approach to quantifying ERß using older mouse ovarian surface epithelium, where ERß is expressed at lower levels than ERα and is therefore harder to detect. We use an antibody highly specific to the ERß1 isoform, together with immunofluorescence, confocal microscopy, and imaging and statistical software to achieve clear, reproducible, and unbiased quantitation of ERß.


Assuntos
Receptor beta de Estrogênio/análise , Regulação da Expressão Gênica , Imuno-Histoquímica/métodos , Microscopia Confocal/métodos , Ovário/metabolismo , Epitélio/metabolismo , Receptor beta de Estrogênio/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Microscopia Intravital/métodos , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/metabolismo
6.
Eur J Histochem ; 63(2)2019 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-31113192

RESUMO

Biomarkers may hold the key towards development and improvement of personalized cancer treatment. For instance, tumour expression of immune system-related proteins may reveal the tumour immune status and, accordingly, determine choice for type of immunotherapy. Therefore, objective evaluation of tumour biomarker expression is needed but often challenging. For instance, human leukocyte antigen (HLA) class I tumour epithelium expression is cumbersome to quantify by eye due to its presence on both tumour epithelial cells and tumour stromal cells, as well as tumour-infiltrating immune cells. In this study, we solved this problem by setting up an immunohistochemical (IHC) double staining using a tissue microarray (TMA) of rectal tumours wherein HLA class I expression was coloured with a blue chromogen, whereas non-epithelial tissue was visualized with a brown chromogen. We subsequently developed a semi-automated image analysis method that identified tumour epithelium as well as the percentage of HLA class I-positive tumour epithelium. Using this technique, we compared HCA2/HC10 and EMR8-5 antibodies for the assessment of HLA class I tumour expression and concluded that EMR8-5 is the superior antibody for this purpose. This IHC double staining can in principle be used for scoring of any biomarker expressed by tumour epithelium.


Assuntos
Antígenos de Histocompatibilidade Classe I/metabolismo , Neoplasias Retais/metabolismo , Animais , Anticorpos Monoclonais Murinos/imunologia , Biomarcadores Tumorais/imunologia , Biomarcadores Tumorais/metabolismo , Epitélio/metabolismo , Antígenos de Histocompatibilidade Classe I/imunologia , Humanos , Imuno-Histoquímica/métodos , Camundongos , Coelhos , Neoplasias Retais/patologia , Análise Serial de Tecidos
7.
Med Sci Monit ; 25: 3366-3373, 2019 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-31062766

RESUMO

BACKGROUND The tripartite motif-containing protein 59 (TRIM59) is an important member of the TRIM family, which regulates biological processes. However, the relationship between TRIM59 and epithelial ovarian cancer (EOC) is not clear. MATERIAL AND METHODS The TRIM59 expression level was detected in EOC tissues and cell lines. CCK-8 assay, Transwell assay, and wound healing assay were performed to determine the effects of TRIM59 on EOC cell proliferation, invasion, and migration. Silencing of the expression of TRIM59 in EOC cells and expression of FAK/AKT/MMP pathway-related protein were detected by Western blot analysis. RESULTS Through bioinformatics analysis, TRIM59 was found to be highly expressed in EOC and was correlated with prognosis of patients. TRIM59 was upregulated in EOC tissues and cells. Silencing TRIM59 significantly suppressed EOC cell proliferation, migration, and invasion. In terms of molecular mechanism, silencing TRIM59 inhibited the FAK/AKT/MMP pathway. CONCLUSIONS TRIM59 is a biomarker for the prognosis of EOC. It is also oncogenic and a potential target for EOC therapy.


Assuntos
Carcinoma Epitelial do Ovário/metabolismo , Quinase 1 de Adesão Focal/metabolismo , Proteínas de Membrana/metabolismo , Metaloproteínas/metabolismo , Neoplasias Ovarianas/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Carcinoma Epitelial do Ovário/genética , Carcinoma Epitelial do Ovário/patologia , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Progressão da Doença , Epitélio/metabolismo , Feminino , Humanos , Metaloproteinases da Matriz/metabolismo , Proteínas de Membrana/genética , Metaloproteínas/genética , Invasividade Neoplásica , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/patologia , Prognóstico , Transdução de Sinais
8.
Int J Mol Sci ; 20(8)2019 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-31014010

RESUMO

Bile acids are a family of amphipathic compounds predominantly known for their role in solubilizing and absorbing hydrophobic compounds (including liposoluble vitamins) in the intestine. Bile acids also are key signaling molecules and inflammatory agents that activate transcriptional factors and cell signaling pathways that regulate lipid, glucose, and energy metabolism in various human disorders, including chronic liver diseases. However, in the last decade increased awareness has been founded on the physiological and chemical heterogeneity of this category of compounds and their possible beneficial or injurious effects on the biliary tree. In this review, we provide an update on the current understanding of the molecular mechanism involving bile acid and biliary epithelium. The last achievements of the research in this field are summarized, focusing on the molecular aspects and the elements with relevance regarding human liver diseases.


Assuntos
Ácidos e Sais Biliares/farmacologia , Epitélio/efeitos dos fármacos , Animais , Sistema Biliar/metabolismo , Epitélio/metabolismo , Humanos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Receptores Acoplados a Proteínas-G/metabolismo , Receptores de Lisoesfingolipídeo/metabolismo , Transdução de Sinais
9.
Int J Biol Macromol ; 132: 801-810, 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-30953722

RESUMO

Iron is essential in fundamental bioactivities, so it makes sense to improve the efficiency of iron on epithelial transport. In this work, a novel polysaccharide­iron(III) complex (FVP-Fe(III)) was prepared from Flammulina velutipes with a specific structure. The FVP-Fe(III) had a molecular weight of 15.13 kDa with a monosaccharide composition of mannose, galacturonic acid, glucose, galactose, xylose, fucose in a molar ratio 3.6:2.1:60.8:18.7:3.9:10.9. In the vitro digestion model, the complex could maintain better solubility and steady of iron than FeSO4. In the cell assay, FVP-Fe(III) showed lower cytotoxicity and better absorption. The transport amount of FVP-Fe(III) was 1.5-fold of FeSO4 at same concentration and 1.8-fold of FeSO4 at same time. The transport was mediated by the peptide transporter (pepT1) active transport pathway and the efflux of the sample was mainly mediated by multidrug-resistance proteins (MRP) transporter. The results of this study suggested that the polysaccharide obtained from F. velutipes could be developed a new kind of iron delivery for further study.


Assuntos
Absorção Fisico-Química , Complexos de Coordenação/química , Complexos de Coordenação/metabolismo , Flammulina/química , Ferro/química , Polissacarídeos/química , Transporte Biológico , Células CACO-2 , Sobrevivência Celular/efeitos dos fármacos , Complexos de Coordenação/toxicidade , Digestão , Epitélio/metabolismo , Humanos , Intestinos/fisiologia , Estômago/fisiologia
10.
Life Sci ; 227: 129-136, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-31002922

RESUMO

Oral epithelial barrier consists of closely controlled structure of the stratified squamous epithelium, which is the gateway to human bodies and encounters a huge burden of microbial, airborne and dietary antigens, as well as masticatory damage. Once this barrier is destroyed, it will trigger bone loss, tissue damage and microbial dysbiosis and lead to diseases, such as periodontitis, oral mucosal diseases and oral cancer. Recently, increasing evidences showed that different factors including microorganism, saliva, proteins and immune components have been considered to play a critical role in the disruption of oral epithelial barrier. Herein, we discussed mechanisms governing the maintenance of oral epithelial barrier. Besides, the role of oral epithelial barrier failure in oral carcinogenesis will also be talked about.


Assuntos
Epitélio/fisiologia , Mucosa Bucal/fisiologia , Boca/fisiologia , Animais , Disbiose , Células Epiteliais/fisiologia , Epitélio/metabolismo , Humanos , Boca/imunologia , Mucosa Bucal/imunologia , Mucosa Bucal/metabolismo , Neoplasias Bucais , Periodontite , Saliva
11.
Int J Mol Sci ; 20(6)2019 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-30897851

RESUMO

BACKGROUND: Many food components influence intestinal epithelial barrier properties and might therefore also affect susceptibility to the development of food allergies. Such allergies are triggered by increased antibody production initiated in Peyer's patches (PP). Usually, the presentation of antigens in the lumen of the gut to the immune cells of the PP is strongly regulated by the follicle-associated epithelium (FAE) that covers the PP. As the food component caprate has been shown to impede barrier properties in villous epithelium, we hypothesized that caprate also affects the barrier function of the PP FAE, thereby possibly contributing a risk factor for the development of food allergies. METHODS: In this study, we have focused on the effects of caprate on the barrier function of PP, employing in vitro and ex vivo experimental setups to investigate functional and molecular barrier properties. Incubation with caprate induced an increase of transepithelial resistance, and a marked increase of permeability for the paracellular marker fluorescein in porcine PP to 180% of control values. These effects are in accordance with changes in the expression levels of the barrier-forming tight junction proteins tricellulin and claudin-5. CONCLUSIONS: This barrier-affecting mechanism could be involved in the initial steps of a food allergy, since it might trigger unregulated contact of the gut lumen with antigens.


Assuntos
Epitélio/metabolismo , Nódulos Linfáticos Agregados/metabolismo , Animais , Linhagem Celular , Claudinas/metabolismo , Immunoblotting , Proteína 2 com Domínio MARVEL/metabolismo , Suínos , Proteínas de Junções Íntimas/metabolismo , Junções Íntimas/metabolismo
12.
Proc Natl Acad Sci U S A ; 116(12): 5362-5369, 2019 03 19.
Artigo em Inglês | MEDLINE | ID: mdl-30837316

RESUMO

Lipid nanovesicles are widely present as transport vehicles in living organisms and can serve as efficient drug delivery vectors. It is known that the size and surface charge of nanovesicles can affect their diffusion behaviors in biological hydrogels such as mucus. However, how temperature effects, including those of both ambient temperature and phase transition temperature (T m), influence vehicle transport across various biological barriers outside and inside the cell remains unclear. Here, we utilize a series of liposomes with different T m as typical models of nanovesicles to examine their diffusion behavior in vitro in biological hydrogels. We observe that the liposomes gain optimal diffusivity when their T m is around the ambient temperature, which signals a drastic change in the nanovesicle rigidity, and that liposomes with T m around body temperature (i.e., ∼37 °C) exhibit enhanced cellular uptake in mucus-secreting epithelium and show significant improvement in oral insulin delivery efficacy in diabetic rats compared with those with higher or lower T m Molecular-dynamics (MD) simulations and superresolution microscopy reveal a temperature- and rigidity-mediated rapid transport mechanism in which the liposomes frequently deform into an ellipsoidal shape near the phase transition temperature during diffusion in biological hydrogels. These findings enhance our understanding of the effect of temperature and rigidity on extracellular and intracellular functions of nanovesicles such as endosomes, exosomes, and argosomes, and suggest that matching T m to ambient temperature could be a feasible way to design highly efficient nanovesicle-based drug delivery vectors.


Assuntos
Hidrogéis/administração & dosagem , Hidrogéis/química , Lipídeos/química , Nanopartículas/química , Animais , Transporte Biológico/efeitos dos fármacos , Diabetes Mellitus Experimental/tratamento farmacológico , Difusão/efeitos dos fármacos , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos/métodos , Epitélio/metabolismo , Insulina/administração & dosagem , Insulina/química , Lipossomos/química , Masculino , Transição de Fase/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Temperatura Ambiente
13.
Nature ; 567(7748): 405-408, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30867598

RESUMO

Loss-of-function mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) compromise epithelial HCO3- and Cl- secretion, reduce airway surface liquid pH, and impair respiratory host defences in people with cystic fibrosis1-3. Here we report that apical addition of amphotericin B, a small molecule that forms unselective ion channels, restored HCO3- secretion and increased airway surface liquid pH in cultured airway epithelia from people with cystic fibrosis. These effects required the basolateral Na+, K+-ATPase, indicating that apical amphotericin B channels functionally interfaced with this driver of anion secretion. Amphotericin B also restored airway surface liquid pH, viscosity, and antibacterial activity in primary cultures of airway epithelia from people with cystic fibrosis caused by different mutations, including ones that do not yield CFTR, and increased airway surface liquid pH in CFTR-null pigs in vivo. Thus, unselective small-molecule ion channels can restore host defences in cystic fibrosis airway epithelia via a mechanism that is independent of CFTR and is therefore independent of genotype.


Assuntos
Fibrose Cística/metabolismo , Epitélio/metabolismo , Canais Iônicos/metabolismo , Mucosa Respiratória/metabolismo , Sistema Respiratório/metabolismo , Anfotericina B/farmacologia , Animais , Bicarbonatos/metabolismo , Células Cultivadas , Fibrose Cística/genética , Regulador de Condutância Transmembrana em Fibrose Cística/deficiência , Regulador de Condutância Transmembrana em Fibrose Cística/genética , Regulador de Condutância Transmembrana em Fibrose Cística/metabolismo , Células Epiteliais/citologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Epitélio/efeitos dos fármacos , Feminino , Humanos , Concentração de Íons de Hidrogênio , Masculino , Mucosa Respiratória/efeitos dos fármacos , Sistema Respiratório/efeitos dos fármacos , ATPase Trocadora de Sódio-Potássio/metabolismo , Suínos
14.
Microb Pathog ; 131: 81-86, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30910720

RESUMO

This study aimed to investigate the mechanism of lipopolysaccharide (LPS) released in the rumen on epithelium barrier function of goats fed a HC diet. Twelve Boer goats were randomly divided into two groups: low-concentrate(LC) diet and high-concentrate(HC) diet treatment. We found that the pH of rumen fluid in the HC group was lower than in the LC group (P < 0.05). The mRNA and protein expression levels of p38 mitogen-activated protein kinase (MAPK), extracellular regulated protein kinases (ERK), and c-Jun N-terminal kinase (JNK) in the rumen epithelium were lower in the LC group than the HC group (P < 0.05). Gene expression and protein levels of the tight junction proteins claudin-1, claudin-4, occludin, and Zona occludin-1 were all greater in the LC group than the HC group (P < 0.05). Staining of claudin-1, occludin and ZO-1 was became irregular. In conclusion, high concentrate diet feeding can impair rumen epithelium function and decrease tight junction protein expression through MAPK signaling pathway.


Assuntos
Dieta/veterinária , Epitélio/metabolismo , Lipopolissacarídeos/metabolismo , Rúmen/metabolismo , Ração Animal/análise , Animais , Claudina-1/genética , Claudina-1/metabolismo , Claudina-4/genética , Claudina-4/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Regulação da Expressão Gênica , Cabras , Concentração de Íons de Hidrogênio , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Ocludina/genética , Ocludina/metabolismo , Distribuição Aleatória , Transdução de Sinais , Proteínas de Junções Íntimas/genética , Proteínas de Junções Íntimas/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
16.
Int J Mol Sci ; 20(6)2019 Mar 16.
Artigo em Inglês | MEDLINE | ID: mdl-30884865

RESUMO

The Hedgehog (Hh) pathway has regulatory roles in maintaining and restoring lingual taste organs, the papillae and taste buds, and taste sensation. Taste buds and taste nerve responses are eliminated if Hh signaling is genetically suppressed or pharmacologically inhibited, but regeneration can occur if signaling is reactivated within the lingual epithelium. Whereas Hh pathway disruption alters taste sensation, tactile and cold responses remain intact, indicating that Hh signaling is modality-specific in regulation of tongue sensation. However, although Hh regulation is essential in taste, the basic biology of pathway controls is not fully understood. With recent demonstrations that sonic hedgehog (Shh) is within both taste buds and the innervating ganglion neurons/nerve fibers, it is compelling to consider Hh signaling throughout the tongue and taste organ cell and tissue compartments. Distinctive signaling centers and niches are reviewed in taste papilla epithelium, taste buds, basal lamina, fibroblasts and lamellipodia, lingual nerves, and sensory ganglia. Several new roles for the innervation in lingual Hh signaling are proposed. Hh signaling within the lingual epithelium and an intact innervation each is necessary, but only together are sufficient to sustain and restore taste buds. Importantly, patients who use Hh pathway inhibiting drugs confront an altered chemosensory world with loss of taste buds and taste responses, intact lingual touch and cold sensation, and taste recovery after drug discontinuation.


Assuntos
Epitélio/metabolismo , Proteínas Hedgehog/genética , Percepção Gustatória/genética , Paladar/genética , Proteínas Hedgehog/metabolismo , Humanos , Sensação/genética , Sensação/fisiologia , Transdução de Sinais/genética , Células Estromais/metabolismo , Paladar/fisiologia , Papilas Gustativas/metabolismo , Papilas Gustativas/fisiologia , Percepção Gustatória/fisiologia , Língua/inervação , Língua/fisiologia
17.
Nat Commun ; 10(1): 1194, 2019 03 18.
Artigo em Inglês | MEDLINE | ID: mdl-30886143

RESUMO

Chronic infections of the fallopian tubes with Chlamydia trachomatis (Ctr) cause scarring and can lead to infertility. Here we use human fallopian tube organoids and genital Ctr serovars D, K and E for long-term in vitro analysis. The epithelial monolayer responds with active expulsion of the bacteria into the lumen and with compensatory cellular proliferation-demonstrating a role of epithelial homeostasis in the defense against this pathogen. In addition, Ctr infection activates LIF signaling, which we find to be an essential regulator of stemness in the organoids. Infected organoids exhibit a less differentiated phenotype with higher stemness potential, as confirmed by increased organoid forming efficiency. Moreover, Ctr increases hypermethylation of DNA, which is an indicator of accelerated molecular aging. Thus, the chronic organoid infection model suggests that Ctr has a long-term impact on the epithelium. These heritable changes might be a contributing factor in the development of tubal pathologies, including the initiation of high grade serous ovarian cancer.


Assuntos
Infecções por Chlamydia/genética , Chlamydia trachomatis/imunologia , Ilhas de CpG/genética , Metilação de DNA/imunologia , Interações entre Hospedeiro e Microrganismos/genética , Células-Tronco/metabolismo , Fatores Etários , Infecções por Chlamydia/imunologia , Infecções por Chlamydia/microbiologia , Chlamydia trachomatis/genética , Doença Crônica , Ilhas de CpG/imunologia , Cistadenocarcinoma Seroso/genética , Cistadenocarcinoma Seroso/imunologia , Cistadenocarcinoma Seroso/microbiologia , Epigênese Genética/genética , Epigênese Genética/imunologia , Epitélio/imunologia , Epitélio/metabolismo , Epitélio/microbiologia , Tubas Uterinas/imunologia , Tubas Uterinas/metabolismo , Tubas Uterinas/microbiologia , Feminino , Interações entre Hospedeiro e Microrganismos/imunologia , Humanos , Microscopia Intravital , Microscopia Confocal , Organoides/imunologia , Organoides/metabolismo , Organoides/microbiologia , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/imunologia , Neoplasias Ovarianas/microbiologia , Sorogrupo , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Análise de Célula Única , Células-Tronco/imunologia , Células-Tronco/microbiologia , Técnicas de Cultura de Tecidos
18.
Cornea ; 38(6): 698-705, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30844839

RESUMO

PURPOSE: To investigate the expression levels of inflammatory cytokines in the conjunctival epithelium and correlations with clinical parameters in dry eye disease (DED). METHODS: This study evaluated 28 patients with Sjögren syndrome (SS) DED, 28 patients with non-SS DED, and 10 controls. The messenger ribonucleic acid (mRNA) expression of inflammatory cytokines such as tumor necrosis factor-α, interleukin (IL)-1ß, IL-6, interferon (IFN)-γ, IL-17, and matrix metalloproteinase 9 (MMP9) from conjunctival epithelium was investigated by real-time polymerase chain reaction. Protein expression was confirmed by immunofluorescence staining. Correlations were evaluated between the mRNA expression of inflammatory cytokines and clinical DED parameters such as ocular surface disease index score, Schirmer I value, tear film breakup time, and corneal and conjunctival staining scores. RESULTS: Patients with non-SS DED expressed significantly more IFN-γ, IL-6, and MMP9 genes in the conjunctival epithelium than the controls (P < 0.05), and all cytokine gene expression was significantly higher in patients with SS DED than in the controls (P < 0.01). Tumor necrosis factor-α, IL-1ß, IL-6, and IL-17 gene expression was higher in patients with SS DED than in the non-SS DED group (P < 0.05). Immunofluorescence staining of conjunctival epithelium demonstrated that positive cells with IL-6 or MMP9 were significantly higher in non-SS DED than in controls (P < 0.01) and much higher in SS DED than in non-SS DED (P < 0.05). Conjunctival staining scores significantly correlated with the expression of IFN-γ, IL-6, IL-17, and MMP9 in both DED groups (P < 0.05 in non-SS DED and P < 0.01 in SS-DED). Interestingly, correlation coefficients of all cytokines were much higher in SS DED compared to non-SS DED. Corneal staining scores showed positive correlations with IFN-γ, IL-17, and MMP9 (P < 0.05), and correlation coefficients were lower than those of conjunctival staining scores. CONCLUSIONS: Conjunctival staining scores may be useful to measure ocular surface inflammation in SS and non-SS DED.


Assuntos
Túnica Conjuntiva/metabolismo , Edema da Córnea/metabolismo , Citocinas/metabolismo , Síndromes do Olho Seco/metabolismo , Síndrome de Sjogren/metabolismo , Adulto , Idoso , Estudos de Casos e Controles , Estudos Transversais , Epitélio/metabolismo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem
19.
Cell Commun Signal ; 17(1): 18, 2019 02 27.
Artigo em Inglês | MEDLINE | ID: mdl-30813930

RESUMO

BACKGROUND: Oral lichen planus (OLP) is known as a chronic inflammatory disease. Our recent studies have suggested that vitamin D/vitamin D receptor (VDR) signaling exerts its protective effects on oral keratinocyte apoptosis by regulating microRNA-802 and p53-upregulated modulator of apoptosis (PUMA), but its roles in oral epithelial inflammatory responses in OLP are still unknown. Herein, we identify lipopolysaccharide (LPS) is able to enhance interferon gamma (IFNγ) and interleukin-1 beta (IL-1ß) productions in human oral keratinocytes (HOKs) dependent on hypoxia-inducible factor-1α (HIF-1α). METHODS: HIF-1α and cytokines levels in HOKs were investigated by real-time PCR and western blotting after LPS challenge. The effects of 1,25(OH)2D3 on LPS-induced HIF-1α and cytokines were tested by real-time PCR, western blotting, siRNA-interference and plasmids transfection techniques. The roles of 1,25(OH)2D3 in regulating HIF-1α levels were investigated using western blotting, siRNA-interference, plasmids transfection and Chromatin Immunoprecipitation (ChIP) assays. Finally, HIF-1α, IFNγ and IL-1ß expressions in oral epithelia derived from mice and individuals were measured by real-time PCR, western blotting and immunohistochemical staining. RESULTS: As a critical regulator, vitamin D suppresses LPS-induced HIF-1α to block IFNγ and IL-1ß productions. Mechanistically, vitamin D inactivates nuclear factor-κB (NF-κB) pathway and up-regulates von Hippel-Lindau (VHL) levels, leading to HIF-1α reduction. Moreover, HIF-1α status of oral epithelia is elevated in VDR-/- mie as well as in VDR-deficient human biopsies, accompanied with increased IFNγ and IL-1ß. CONCLUSION: Collectively, this study uncovers an unrecognized roles of vitamin D/VDR signaling in regulating cytokines in oral keratinocytes and reveals the molecular basis of it.


Assuntos
Epitélio/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Interferon gama/metabolismo , Interleucina-1beta/metabolismo , Lipopolissacarídeos/farmacologia , Receptores de Calcitriol/metabolismo , Transdução de Sinais , Vitamina D/metabolismo , Animais , Sequência de Bases , Calcitriol/farmacologia , Linhagem Celular , Epitélio/efeitos dos fármacos , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Queratinócitos/efeitos dos fármacos , Queratinócitos/metabolismo , Líquen Plano Bucal/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Boca/metabolismo , NF-kappa B/metabolismo , Regulação para Cima/efeitos dos fármacos , Proteína Supressora de Tumor Von Hippel-Lindau/metabolismo
20.
Biomater Sci ; 7(5): 1949-1961, 2019 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-30793722

RESUMO

The application of conventional approaches to diabetic wound regeneration has some limitations. Thus, skin substitutes could be a new therapeutic possibility. In this regard, fibrin scaffolds are promising materials due to their desirable characteristics. Since defective fibroblasts caused by diabetes can disrupt regeneration, it seems that the use of living cells can improve the healing process. Thus, based on this fact, a cellular fibrin membrane was used to evaluate the diabetic wound healing in rats. The fibrin membrane was fabricated using fresh frozen plasma on which isolated fibroblasts were cultured. The wound model was created on 36 diabetic rats that were randomly divided into three groups: control, membrane, and cellular fibrin membrane (CM). Wound photogramography and immuno-histopathological staining were performed during consecutive days after treatment. Macroscopic evaluation of the wounds indicated a noteworthy enhancement of wound closure in the CM group. In the CM group, the re-epithelialization rate on day 7, 10 (p < 0.001), and 14 (p < 0.05), the fibroblast percentage on day 3 (p < 0.01) and 7 (p < 0.05) and the collagenization in all days were significantly higher than those of other groups (p < 0.001). The fibroblast number in the CM group on day 10 was significantly (p < 0.01) lower than that in the other groups. Contrary to the neutrophil and angiogenesis percentages that had no significant difference among the groups at different points of time (p > 0.05), the macrophage percentage on day 7 (P < 0.01), 10, and 14 (p < 0.05) was significantly lower in the CM group as compared to that in other groups. Overall, it seems that the use of a fibroblast-loaded fibrin membrane is an attractive strategy to promote diabetic wound healing.


Assuntos
Células Alógenas/citologia , Materiais Biocompatíveis/farmacologia , Diabetes Mellitus Experimental/fisiopatologia , Fibroblastos/citologia , Membranas Artificiais , Cicatrização/efeitos dos fármacos , Animais , Materiais Biocompatíveis/metabolismo , Contagem de Células , Colágeno/metabolismo , Epitélio/efeitos dos fármacos , Epitélio/metabolismo , Fibrina/metabolismo , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Masculino , Neovascularização Fisiológica/efeitos dos fármacos , Neutrófilos/citologia , Neutrófilos/efeitos dos fármacos , Ratos , Ratos Wistar , Pele/irrigação sanguínea , Pele/efeitos dos fármacos , Pele/imunologia , Pele/patologia
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