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1.
Gene ; 710: 218-232, 2019 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-31158448

RESUMO

Alterations in the global gene expression profile are considered to contribute to the various physiological and pathological changes during the course of ageing. Genes that code for the molecular components of the innate system are alter markedly as ageing occurs; and this may define the susceptibility of very young and very old individuals to reproductive tract infections. The expression pattern of genes that code for beta-defensins (effectors of innate immune response) in male reproductive tract tissues of different stages of ageing is not yet reported. Further, the induction of beta-defensins during endotoxin challenge and whether epigenetic modulators can influence the expression of these genes in different stages of ageing are not reported. We analyzed the basal mRNA levels of beta-defensins and defensin-like proteins (Sperm Associated Antigen 11 (SPAG11) family members), their induction during endotoxin challenge and modulation by epigenetic modifiers (Trichostatin A and Azacytidine) in the caput, cauda, testis, prostate and seminal vesicle of rats that represent early stage to late stages of life (20 day to 730 day old). We observed differential basal gene expression pattern in the male reproductive tract tissues and the induction by LPS was not consistent neither among the age groups not the tissues analyzed. Trichostatin A and Azacytidine also influenced antimicrobial gene expression and the pattern was not consistent in different tissues obtained from different age groups. Results of this study demonstrate that antimicrobial gene expression varies to a great extent during ageing and is strongly influenced by endotoxins and epigenetic modulators.


Assuntos
Envelhecimento/genética , Genitália Masculina/química , Glicopeptídeos/genética , beta-Defensinas/genética , Animais , Azacitidina/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Genitália Masculina/efeitos dos fármacos , Ácidos Hidroxâmicos/farmacologia , Lipopolissacarídeos/farmacologia , Masculino , Ratos , Ratos Wistar
2.
J Dairy Sci ; 102(6): 5706-5712, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30954263

RESUMO

Antimicrobial peptides are a common defense against bacterial infections in many species and a significant part of the innate immune response of the bovine mammary gland. The objective of this study was to investigate the influence of epigenetic factors on vitamin D and toll-like receptor-mediated induction of ß-defensins in mammary epithelial cells. Primary bovine mammary epithelial cells were treated with lipopolysaccharide (LPS, 0 or 100 ng/mL), 1,25-dihydroxyvitamin D3 [1,25(OH)2D3, 0 or 10 nM], and 5-aza-2'-deoxycytidine (5-Aza, inhibitor of DNA methyltransferase, 0 or 5 µM) or trichostatin A (TSA, inhibitor of histone deacetylase, 0 or 80 nM) in a factorial arrangement. Effects of treatments on ß-defensin gene expression along with genes for cytokines and enzymes known to be induced by LPS or 1,25(OH)2D3 were evaluated by quantitative PCR. The LPS treatment induced expression of ß-defensin (DEFB)3, DEFB5, DEFB7, DEFB10, enteric ß-defensin (EBD), lingual antimicrobial peptide (LAP), and tracheal antimicrobial peptide (TAP); whereas, the 1,25(OH)2D3 treatment increased DEFB5 and DEFB7 expression and decreased LAP. The 5-Aza treatment increased expression of DEFB3, DEFB5, DEFB10, EBD, LAP, and TAP in the presence and absence of LPS. The TSA treatment increased expression of DEFB3, DEFB4, DEFB5, DEFB7, and DEFB10 in the absence of LPS but decreased LPS-induced expression of and LAP and TAP. Together these results indicate that ß-defensin expression in bovine mammary epithelial cells is likely influenced by DNA methylation and histone acetylation. Investigation of environmental and nutritional factors that influence epigenetic control of ß-defensins in the mammary gland may be beneficial for improving resistance to intramammary infections.


Assuntos
Bovinos/metabolismo , Células Epiteliais/metabolismo , Histona Desacetilases/metabolismo , Lipopolissacarídeos/metabolismo , Glândulas Mamárias Animais/metabolismo , Metiltransferases/metabolismo , Vitamina D/análogos & derivados , beta-Defensinas/genética , Animais , Bovinos/genética , Metilação de DNA , Feminino , Histona Desacetilases/genética , Glândulas Mamárias Animais/citologia , Metiltransferases/genética , Vitamina D/metabolismo , beta-Defensinas/metabolismo
3.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 36(4): 371-375, 2019 Apr 10.
Artigo em Chinês | MEDLINE | ID: mdl-30950029

RESUMO

OBJECTIVE: To investigate the association between the polymorphisms of 5'-UTR -52G/A (rs1799946), -44C/G (rs1800972), -20G/A (rs11362) in DEFB1 gene with chronic periodontitis in Henan Han population. METHODS: Peripheral blood genomic DNA of 436 patients with chronic periodontitis and 440 healthy controls were extracted and subjected to PCR-Sanger sequencing to determine the genotypes of DEFB1 5'-UTR -52G/A (rs1799946), -44C/G (rs1800972) and -20G/A (rs11362). The distribution of genotypes, allele frequencies and risk factors were analyzed by chi-square test and Logistic regression. RESULTS: There was no significant difference between healthy controls and chronic periodontitis in the genotype of -52G/A PCR- (rs1799946) and -20G/A (rs11362) (P> 0.05). While a significant difference was found between healthy controls and chronic periodontitis in -44C/G (rs1800972), the CC and CG genotype rate in the two groups were 64.5%, 82.1% and 28.2%, 14.4% respectively. One-way logistic analysis showed that the CG, GG genotype and allele G might be a protective factor. CONCLUSION: The DEFB1 -44C/G (rs1800972) is associated with chronic periodontitis in Henan Han population, and the -44CG, GG genotype and G allele may be the protective factors of chronic periodontitis in Henan Han population.


Assuntos
Periodontite Crônica , Polimorfismo Genético , beta-Defensinas/genética , Alelos , Estudos de Casos e Controles , Periodontite Crônica/genética , Frequência do Gene , Predisposição Genética para Doença , Genótipo , Humanos
4.
Nat Commun ; 10(1): 1305, 2019 03 21.
Artigo em Inglês | MEDLINE | ID: mdl-30899005

RESUMO

Failure to predict and understand the causes of preterm birth, the leading cause of neonatal morbidity and mortality, have limited effective interventions and therapeutics. From a cohort of 2000 pregnant women, we performed a nested case control study on 107 well-phenotyped cases of spontaneous preterm birth (sPTB) and 432 women delivering at term. Using innovative Bayesian modeling of cervicovaginal microbiota, seven bacterial taxa were significantly associated with increased risk of sPTB, with a stronger effect in African American women. However, higher vaginal levels of ß-defensin-2 lowered the risk of sPTB associated with cervicovaginal microbiota in an ethnicity-dependent manner. Surprisingly, even in Lactobacillus spp. dominated cervicovaginal microbiota, low ß-defensin-2 was associated with increased risk of sPTB. These findings hold promise for diagnostics to accurately identify women at risk for sPTB early in pregnancy. Therapeutic strategies could include immune modulators and microbiome-based therapeutics to reduce this significant health burden.


Assuntos
Colo do Útero/microbiologia , Imunidade Inata , Microbiota/imunologia , Nascimento Prematuro/diagnóstico , Vagina/microbiologia , beta-Defensinas/genética , Adulto , Grupo com Ancestrais do Continente Africano , Teorema de Bayes , Biomarcadores/metabolismo , Estudos de Casos e Controles , Grupo com Ancestrais do Continente Europeu , Feminino , Expressão Gênica , Humanos , Recém-Nascido , Lactobacillus/classificação , Lactobacillus/imunologia , Lactobacillus/isolamento & purificação , Mobiluncus/classificação , Mobiluncus/imunologia , Mobiluncus/isolamento & purificação , Gravidez , Nascimento Prematuro/etnologia , Nascimento Prematuro/imunologia , Nascimento Prematuro/fisiopatologia , Prognóstico , Risco , beta-Defensinas/imunologia
5.
Poult Sci ; 98(7): 3022-3028, 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-30915472

RESUMO

Sustained production of good quality eggs for longer production cycles is a challenge for poultry farms. The impact of aging on the mucosal immune defense in the isthmus and uterus of hens, where the eggshell membrane and eggshell are formed, remains obscure. Thus, the aim of this study was to determine whether aging affects the mucosal tight junction (TJ) proteins, the synthesis of antimicrobial peptides including avian ß-defensins (AvBDs) and cathelicidins (CATHs), and Toll-like receptors (TLRs) in the isthmus and uterus of laying hens. Young and aged White Leghorn laying hens (35 and 130 wk old, respectively) were used. Total RNA and protein contents were isolated from the isthmic and uterine mucosae of these hens to examine the expression of TJ proteins, AvBD, and CATH genes and AvBD proteins by the real-time polymerase chain reaction and western blotting. The results showed that the mRNA expression of TJ proteins, namely zonula occludin 2 in the isthmus and occludin in the uterus, was higher in aged hens than in young hens. Expression of 2 AvBD genes in the isthmus and 4 AvBD genes in the uterus was higher in aged hens than in young hens. However, the expression of AvBD proteins 1 and 11 was not altered by aging. Expressions of CATH genes were not affected by aging in the isthmus or uterus. Expression of TLR genes was higher in aged hens than in young hens in the isthmus, while their expression in the uterus was not affected by aging. It can be concluded that aged hens have a higher potential ability to express TJ proteins and AvBDs for mucosal defense in the isthmic and uterine mucosae than in young hens.


Assuntos
Envelhecimento/metabolismo , Galinhas/fisiologia , Imunidade Inata/imunologia , Útero/metabolismo , Animais , Catelicidinas/genética , Catelicidinas/metabolismo , Feminino , Expressão Gênica , Imunidade Inata/genética , Membrana Mucosa , Proteínas de Junções Íntimas/genética , Proteínas de Junções Íntimas/metabolismo , Receptores Toll-Like/genética , Receptores Toll-Like/metabolismo , beta-Defensinas/genética , beta-Defensinas/metabolismo
6.
Mol Genet Genomics ; 294(3): 679-692, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30834967

RESUMO

Cathartidae is a small family of large-bodied carrion-feeding birds, of which the turkey vulture (Cathartes aura, Cathartidae) is the most widespread distributed. To investigate the chemoreception system, detoxification system, and immune system in the turkey vulture, we compared its genome to 14 other avian genomes. Comparative genomics demonstrated the expansion in the chemoreception system, especially the olfactory receptors, while the genes in the detoxification system of the turkey vulture did not show apparent expansion. We identified five positively selected genes associated with the immune system in the turkey vulture, which was likely to strengthen the immune defense against pathogenic invasion. Functional enrichment analysis indicated that many positively selected genes were involved in the regulation of immune system processes, implying important reorganization of the immune system in the turkey vulture. The turkey vulture-specific missense mutations were found in one positively selected gene (BCL6), and all the missense mutations were classified as deleterious by PolyPhen-2, possibly contributing to immune adaptation to the carrion feeding. Furthermore, we identified four turkey vulture-specific missense mutations in three ß-defensin genes of the turkey vulture, which was an indispensable part in the innate immunity (a natural barrier against invasive microbes including bacteria, fungi, and viruses). Our genomic analyses in the turkey vulture provided insights into the genetic signatures of the adaptation to the carrion feeding.


Assuntos
Proteínas Aviárias/genética , Aves/genética , Genoma/genética , Genômica/métodos , Sequência de Aminoácidos , Animais , Bactérias/patogenicidade , Aves/classificação , Aves/microbiologia , Comportamento Alimentar , Fungos/patogenicidade , Sistema Imunitário/metabolismo , Sistema Imunitário/microbiologia , Filogenia , Homologia de Sequência de Aminoácidos , Virulência , Vírus/patogenicidade , beta-Defensinas/genética
7.
Vet Res Commun ; 43(2): 77-89, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30863917

RESUMO

Ovine ruminal epithelial cells (ORECs) not only have a physical barrier function but also can secrete host defence peptides (HDPs), such as sheep ß-defensin-1 (SBD-1). As a feed additive, Saccharomyces cerevisiae can enhance the host's innate immunity. ß-glucan, a cell wall component of Saccharomyces cerevisiae, can stimulate innate immune responses and trigger the up-regulation of SBD-1 in ORECs. The signaling mechanisms involved in ß-glucan-induced SBD-1 expression are not completely understood. The aim of this study was to identify the receptors and intracellular pathways involved in the up-regulation of SBD-1 induced by ß-glucan. ORECs were cultured, and the regulatory mechanisms of ß-glucan-induced up-regulation of SBD-1 were detected using quantitative real-time PCR (qPCR), enzyme-linked immunosorbent assay (ELISA), and western blotting. TLR-2 and MyD88 knockdown or inhibition attenuated ß-glucan-induced SBD-1 expression. We also showed that inhibition of MAPK and NF-κB pathways significantly reduced ß-glucan-induced SBD-1 expression. These results demonstrate that ß-glucan-induced SBD-1 expression is TLR-2-MyD88-dependent and may be regulated by both MAPK and NF-κB pathways. Since NF-κB inhibition had a greater effect on the down-regulation of ß-glucan-induced SBD-1 expression, the NF-κB pathway may be the dominant signaling pathway involved in the regulation of defensin expression. Our studies demonstrate that ß-glucan-induced SBD-1 expression is mediated through the TLR-2-MyD88-NF-κB/MAPK pathway. Our results would contribute to the understanding of immunological modulations in the gastrointestinal tract triggered by probiotic yeast cell wall components.


Assuntos
Células Epiteliais/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno , Fator 88 de Diferenciação Mieloide , NF-kappa B , Receptores Toll-Like , beta-Defensinas/genética , beta-Glucanas/farmacologia , Animais , Células Cultivadas , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/imunologia , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Fator 88 de Diferenciação Mieloide/metabolismo , NF-kappa B/metabolismo , Probióticos/farmacologia , Rúmen/efeitos dos fármacos , Saccharomyces cerevisiae/química , Ovinos , Receptores Toll-Like/metabolismo
8.
Medicine (Baltimore) ; 98(5): e14131, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30702566

RESUMO

OBJECTIVE: The 1st exon 5' noncoding region rs1799946 (-52A/G), rs1800972 (-44C/G), rs11362 (-20A/G) 3 single-nucleotide polymorphisms (SNPs) on human ß-defensin-1 (HBD-1) gene affect its transcription and posttranscriptional mRNA stability then affect the activity of HBD-1. This study was to investigate the effects of HBD-1 gene rs1799946, rs1800972, and rs11362 locus SNPs on genetic susceptibility and prognosis of acute respiratory distress syndrome (ARDS). METHODS: A total of 300 patients with ARDS (ARDS group) and 240 patients who were admitted to the intensive care unit and had a high risk of ARDS but did not progress to ARDS (control group) were included in this study. The genotypes of HBD-1 gene rs1799946, rs1800972, and rs11362 locus and serum HBD-1 were detected. Patients were followed for 60 days with development of ARDS as a primary outcome, ARDS-related mortality and organ dysfunction were secondary outcomes. RESULTS: HBD-1 gene rs1799946 and rs11362 gene mutations were not risk factors for ARDS (P > .05). Mutation allele G of rs1800972 locus in HBD-1 gene was a risk factor for ARDS. There was no significant difference in serum HBD-1 levels between patients with different genotypes of rs1799946 and rs11362 locus in the HBD-1 gene (P > .05). HBD-1 gene rs1800972 locus wild type, heterozygous, and mutant homozygous serum levels of HBD-1 gradually decreased, the difference was statistically significant (P < .001). The 60-day survival rate of subjects with wild type, heterozygous, and mutant homozygote at the rs1800972 locus of HBD-1 gene decreased sequentially (81.7%, 48.9%, and 39.7%), and the difference was statistically significant (P < .05). CONCLUSION: The SNP of rs1800972 (-44C/G) in HBD-1 gene is associated with the risk of ARDS. The rs1800972 locus G allele carriers are more likely to develop ARDS and have a poor prognosis.


Assuntos
Síndrome do Desconforto Respiratório do Adulto/genética , beta-Defensinas/genética , Idoso , Biomarcadores , Estudos de Casos e Controles , Feminino , Genótipo , Humanos , Unidades de Terapia Intensiva , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , Prognóstico
9.
Vet Res ; 50(1): 8, 2019 Feb 04.
Artigo em Inglês | MEDLINE | ID: mdl-30717800

RESUMO

The rumen epithelium of sheep serves as an immune interface with the environment and secretes antimicrobial peptides with bactericidal function against various pathogens. Sheep beta-defensin-1 (SBD-1), an antimicrobial peptide, is secreted from ovine ruminal epithelial cells (OREC) in response to microbial infections. Mannan, the main component of the Saccharomyces cerevisiae cell wall can stimulate innate and regulatory immune responses that could improve the gastrointestinal environment. We aimed at investigating the effects of mannan on SBD-1 expression and the downstream signaling pathways stimulated in OREC. We cultured OREC; assessed the effects of mannan on SBD-1 expression by qPCR and ELISA; and then investigated the underlying signaling pathways using qPCR, ELISA, Western blotting, immunohistochemistry, and immunohistofluorescence. Interestingly, mannan markedly upregulated SBD-1 expression in a concentration- and time-dependent manner. Dectin-2 Mouse mAb, Syk specific inhibitor R406, and specific inhibitors of the p38, ERK1/2, JNK, and NF-κB pathways attenuated mannan-induced SBD-1 expression to varying degrees. These results demonstrate that SBD-1 is upregulated by mannan via the Dectin-2-Syk axis, and this is regulated to a large extent through the mitogen-activated protein kinase (MAPK) p38 and less so through the ERK1/2 and JNK or the NF-κB pathway. Our findings highlight the immunomodulatory effects of mannan on OREC in terms of mannan-induced SBD-1 expression.


Assuntos
Peptídeos Catiônicos Antimicrobianos/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Mananas/farmacologia , Carneiro Doméstico/genética , beta-Defensinas/genética , Animais , Células Epiteliais/metabolismo , Lectinas Tipo C/metabolismo , Rúmen/metabolismo , Saccharomyces cerevisiae/química , Carneiro Doméstico/metabolismo , Transdução de Sinais , Quinase Syk/metabolismo , beta-Defensinas/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
10.
J Assist Reprod Genet ; 36(4): 787-797, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30712073

RESUMO

PURPOSE: ß-defensins are antimicrobial peptides expressed at mucosal level of male and female genito-urinary tract, where they exert protective functions against infections, possibly preserving human health and fertility. In our study, we investigated the possible involvement of ß-defensins in female and male infertility in Italian infertile couples (i) evaluating the presence of human ß-defensin 1 (hBD-1) in follicular fluid (FF) and its correlation with in vitro fertilization (IVF) outcomes; (ii) investigating the relationship between hBD-1 levels in semen and IVF outcomes (comprising correlation with sperm parameters); and (iii) exploring the effect of hBD-1 peptide on spermatozoa motility in vitro. METHODS: A perspective observational analytic pilot study was conducted. hBD-1 concentration was measured with ELISA assay in FF and semen from 50 couples that underwent assisted procreation technique procedures due to infertility status. Moreover, hBD-1 exogenous peptide was administered to 29 normozoospermic semen and their motility was recorded. RESULTS: hBD-1 was detected in FF and its levels were significantly higher in women with good fertilization rate (≥ 75%), respect to those with a poor fertilization rate (< 75%). The hBD-1 semen concentrations in oligo-asthenozoospermic subjects were significantly lower than that in normozoospermic men. Instead, hBD-1 level in sperm and FF not correlated with pregnancy rate. Finally, incubation of sperm with exogenous hBD-1 significantly increased progressive motility after 1 h and 24 h. CONCLUSIONS: Being aware of the relatively small sample size and medium power, our results possibly suggest that hBD-1 could influence oocyte and sperm quality, and could improve, when exogenously added, sperm motility.


Assuntos
Fertilidade/genética , Infertilidade Masculina/genética , Motilidade Espermática/genética , beta-Defensinas/genética , Adulto , Feminino , Fertilização In Vitro/métodos , Líquido Folicular/metabolismo , Humanos , Infertilidade Masculina/patologia , Infertilidade Masculina/terapia , Masculino , Oócitos/metabolismo , Projetos Piloto , Gravidez , Taxa de Gravidez , Sêmen/metabolismo , Contagem de Espermatozoides , Espermatozoides/metabolismo , Espermatozoides/patologia , beta-Defensinas/farmacologia
11.
Fish Shellfish Immunol ; 88: 207-216, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30807859

RESUMO

Antimicrobial peptides (AMPs) play essential roles in the innate immune system to protect against a wide variety of pathogens in aquatic environments. In this study, three very important AMPs, cathelicidin, hepcidin and defensin, were identified in the critically endangered Acipenser dabryanus. The full-length cDNA sequences of these three AMPs were identified from transcriptome sequencing and the rapid amplification of cDNA ends (RACE) technique. Phylogenetic analysis showed that cathelicidin formed a clade with the other members of the cathelicidin family, and similar results were obtained for hepcidin. The A. dabryanus ß-defensin belonged to the fish class 2 ß-defensins. A tissue distribution study showed that the three AMP transcripts could be detected constitutively in various tissues. The highest expression levels of cathelicidin and hepcidin were found in the liver, while defensin was primarily expressed in the skin. Bacterial challenge in vivo revealed significant changes in the gene expression of the three AMPs at both mucosal sites and systemic sites. Striking upregulation of cathelicidin and hepcidin was observed in the skin at 12 h post-challenge, with increases of more than 7000-fold and 1000-fold, respectively, compared to the control, and the expression of defensin mRNA was remarkably elevated in the hindgut (by 230-fold at 6 h post-challenge). Moreover, according to the expression profiles of the AMPs post-challenge, we found that the mucosal immune response occurred earlier than the systemic immune response following bacterial infection. Our results suggest that these three novel AMPs may play important roles in the innate immune system of A. dabryanus to protect against invading pathogens, especially during the mucosal immune response.


Assuntos
Peptídeos Catiônicos Antimicrobianos/imunologia , Peptídeos Catiônicos Antimicrobianos/isolamento & purificação , Peixes/imunologia , Animais , Peptídeos Catiônicos Antimicrobianos/genética , DNA Complementar , Edwardsiella tarda , Espécies em Perigo de Extinção , Infecções por Enterobacteriaceae/imunologia , Infecções por Enterobacteriaceae/veterinária , Doenças dos Peixes/imunologia , Doenças dos Peixes/microbiologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Proteínas de Peixes/isolamento & purificação , Peixes/genética , Peixes/microbiologia , Hepcidinas/genética , Hepcidinas/isolamento & purificação , Imunidade Inata , Filogenia , Análise de Sequência de DNA , beta-Defensinas/genética , beta-Defensinas/isolamento & purificação
12.
Dev Comp Immunol ; 95: 89-95, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30753854

RESUMO

The chicken yolk sac (YS) plays an important role in nutrient absorption and immune function for the developing embryo. The avian ß-defensins (AvBD) are cationic peptides that are important members of the innate immune system. The objective of this study was to profile AvBD mRNA expression patterns and distribution of cells expressing AvBD mRNA in the chicken YS. Expression of AvBD1, 2, 7, and 10 mRNA was low at embryonic day 7 (e7), increased to e9 through e13 and then declined to e19. Using in situ hybridization, AvBD10 mRNA was found to be expressed in endodermal epithelial cells, while AvBD1, 2, and 7 mRNA were expressed in heterophils. The developmental expression pattern and distribution of AvBD mRNA in the YS reveals the importance of these genes to protection of the developing chick embryo.


Assuntos
Proteínas Aviárias/genética , Desenvolvimento Embrionário/imunologia , Regulação da Expressão Gênica no Desenvolvimento/imunologia , Saco Vitelino/imunologia , beta-Defensinas/genética , Animais , Proteínas Aviárias/imunologia , Embrião de Galinha , Galinhas , Endoderma/citologia , Endoderma/imunologia , Endoderma/metabolismo , Células Epiteliais/imunologia , Células Epiteliais/metabolismo , RNA Mensageiro/metabolismo , Saco Vitelino/crescimento & desenvolvimento , Saco Vitelino/metabolismo , beta-Defensinas/imunologia
13.
J Prosthodont Res ; 63(2): 162-166, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30529003

RESUMO

PURPOSE: This study aimed to clarify the influence of chewing on human ß-defensin 2 (hBD-2) and secretory immunoglobulin A (SIgA) expression levels. METHODS: We included 15 healthy males with no missing teeth (mean age, 25.5±2.5years). Subjects were instructed to chew a piece of gum for 30min. Saliva and skin-extraction samples were collected before and after chewing for 15 and 30min. hBD-2 and SIgA concentrations in the samples were determined using enzyme-linked immunosorbent assay (ELISA). hBD-2 and SIgA expression levels before and after chewing were analyzed using the Mann-Whitney U test, following the Friedman test. The significance level was 0.05. RESULTS: The hBD-2 level in skin-extraction samples was significantly different before (99.4±17.3pg/mL) and after chewing for 30min (142±23.0pg/mL). The SIgA level in skin-extraction samples was also significantly different before (2.39±0.25µg/mL) and after chewing for 30min (3.61±0.33µg/mL). No significant difference was noted in either hBD-2 or SIgA secretion rate in saliva between before and after chewing. CONCLUSIONS: Chewing gum for 30min increased hBD-2 and SIgA expression levels in skin. Moreover, chewing gum could influence the secretion pattern of these two biomolecules on skin, but not in saliva.


Assuntos
Expressão Gênica , Imunoglobulina A Secretora/genética , Imunoglobulina A Secretora/metabolismo , Mastigação/imunologia , Pele/imunologia , Pele/metabolismo , beta-Defensinas/genética , beta-Defensinas/metabolismo , Adulto , Epitélio/imunologia , Epitélio/metabolismo , Humanos , Masculino , Fatores de Tempo , Adulto Jovem
14.
Mol Genet Genomic Med ; 7(1): e00509, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30549243

RESUMO

BACKGROUND: The aim of this study was to establish the association of two polymorphisms of the ß-defensin 1 gene (DEFB1, OMIM#602056) with the risk of developing type 2 diabetes mellitus (T2DM) in a group of Mexican patients. METHODS: The 5'UTR -20 G/A, and -44 C/G polymorphisms of DEFB1 gene were genotyped by 5' exonuclease TaqMan assays in a group of 252 patients with T2DM and 522 healthy control. RESULTS: Under dominant and additive models adjusted for the risk factors, the C allele of the -44 C/G polymorphism was associated with increased risk of T2DM (OR = 1.63, 95% CI = 1.07-2.48, pCdom  = 0.021 and OR = 1.42, 95% CI = 1.05-1.91, pCadd  = 0.023, respectively). In addition, the linkage disequilibrium analysis showed that AC haplotype was associated with an increased risk of developing T2DM (OR = 4.39, p = 0.04). The in-silico analysis showed that the -44 C allele produces a binding site for the transcription factor Ikaros (IK). CONCLUSION: This study demonstrates that the C allele of -44 C/G polymorphism, as well as haplotype AC are associated with the presence of T2DM in the Mexican population. The variation in this polymorphism of the DEFB1 gene could increase the migration of the macrophages to pancreatic islets accelerate the ß-cell dysfunction in T2DM.


Assuntos
Diabetes Mellitus Tipo 2/genética , Polimorfismo de Nucleotídeo Único , beta-Defensinas/genética , Idoso , Sítios de Ligação , Feminino , Haplótipos , Humanos , Fator de Transcrição Ikaros/metabolismo , Masculino , Pessoa de Meia-Idade , Ligação Proteica , beta-Defensinas/química , beta-Defensinas/metabolismo
15.
Eur J Dermatol ; 28(6): 790-794, 2018 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-30530405

RESUMO

Progranulin (PGRN) is a multi-functional protein known to be involved in diverse biological processes, including tumourigenesis, anti-inflammation, and anti-infection. PGRN expression in sera or tissues is elevated in a variety of malignancies and is associated with poor prognosis. However, it remains to be determined whether PGRN is involved in Mycosis fungoides (MF). To investigate the roles of PGRN in MF. Serum PGRN levels were measured in patients with MF and normal controls by enzyme-linked immunosorbent assay. PGRN expression in MF and normal skin was examined by quantitative reverse transcription polymerase chain reaction and immunohistochemistry. Moreover, we analysed correlations between the expression levels of PGRN and antimicrobial peptides in lesional skin. PGRN levels were significantly lower in sera of MF patients than those of normal controls. PGRN mRNA levels in lesional skin of MF were also significantly decreased. Immunohistochemical staining revealed that PGRN was expressed in epidermal keratinocytes of normal controls, however, PGRN expression in epidermal keratinocytes was also weaker in MF skin. Furthermore, significant inverse correlations were identified between PGRN and antimicrobial peptide mRNA expression. These results suggest that low PGRN expression may contribute to the frequent occurrence of skin infections in patients with MF.


Assuntos
Micose Fungoide/genética , Micose Fungoide/metabolismo , Progranulinas/metabolismo , RNA Mensageiro/metabolismo , Neoplasias Cutâneas/genética , Neoplasias Cutâneas/metabolismo , Adulto , Idoso , Calgranulina A/genética , Estudos de Casos e Controles , Feminino , Humanos , Queratinócitos/metabolismo , Masculino , Pessoa de Meia-Idade , Micose Fungoide/sangue , Micose Fungoide/complicações , Progranulinas/sangue , Progranulinas/genética , Pele/metabolismo , Dermatopatias Infecciosas/etiologia , Neoplasias Cutâneas/sangue , Adulto Jovem , beta-Defensinas/genética
16.
Int J Mol Sci ; 20(1)2018 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-30577574

RESUMO

l-Tryptophan (Trp) is known to play an important role in the health of the large intestine. However, a role of dietary Trp in the small-intestinal mucosal barrier and microbiota remains poorly understood. The present study was conducted with weaned piglets to address this issue. Postweaning piglets were fed for 4 weeks a corn- and soybean meal-based diet supplemented with 0 (Control), 0.1, 0.2, or 0.4% Trp. The small-intestinal microbiota and serum amino acids were analyzed by bacterial 16S rRNA gene-based high-throughput sequencing methods and high-performance liquid chromatography, respectively. The mRNA levels for genes involved in host defense and the abundances of tight-junction proteins in jejunum and duodenum were measured by real time-PCR and Western blot techniques, respectively. The concentrations of Trp in the serum of Trp-supplemented piglets increased in a dose-dependent manner. Compared with the control group, dietary supplementation with 0.2⁻0.4% Trp reduced the abundances of Clostridium sensu stricto and Streptococcus in the jejunum, increased the abundances of Lactobacillus and Clostridium XI (two species of bacteria that can metabolize Trp) in the jejunum, and augmented the concentrations of secretory immunoglobulin A (sIgA) as well as mRNA levels for porcine ß-defensins 2 and 3 in jejunal tissues. Moreover, dietary Trp supplementation activated the mammalian target of rapamycin signaling and increased the abundances of tight-junction proteins (zonula occludens (ZO)-1, ZO-3, and claudin-1) in jejunum and duodenum. We suggested that Trp-metabolizing bacteria in the small intestine of weaned pigs primarily mediated the beneficial effects of dietary Trp on its mucosal integrity, health, and function.


Assuntos
Suplementos Nutricionais , Mucosa Intestinal/metabolismo , Triptofano/metabolismo , Aminoácidos/sangue , Animais , Animais Recém-Nascidos , Biodiversidade , Microbioma Gastrointestinal , Expressão Gênica , Imunoglobulina A Secretora/biossíntese , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/microbiologia , Permeabilidade , Transdução de Sinais , Suínos , Proteínas de Junções Íntimas/genética , Proteínas de Junções Íntimas/metabolismo , Triptofano/farmacologia , Desmame , beta-Defensinas/genética , beta-Defensinas/metabolismo
17.
J Immunol Res ; 2018: 5492941, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30581875

RESUMO

Novel alternatives to antibiotics are needed for the swine industry, given increasing restrictions on subtherapeutic use of antibiotics. Augmenting the synthesis of endogenous host defense peptides (HDPs) has emerged as a promising antibiotic-alternative approach to disease control and prevention. To facilitate the identification of HDP inducers for swine use, we developed a stable luciferase reporter cell line, IPEC-J2/PBD3-luc, through permanent integration of a luciferase reporter gene driven by a 1.1 kb porcine ß-defensin 3 (PBD3) gene promoter in porcine IPEC-J2 intestinal epithelial cells. Such a stable reporter cell line was employed in a high-throughput screening of 148 epigenetic compounds and 584 natural products, resulting in the identification of 41 unique hits with a minimum strictly standardized mean difference (SSMD) value of 3.0. Among them, 13 compounds were further confirmed to give at least a 5-fold increase in the luciferase activity in the stable reporter cell line, with 12 being histone deacetylase (HDAC) inhibitors. Eight compounds were subsequently observed to be comparable to sodium butyrate in inducing PBD3 mRNA expression in parental IPEC-J2 cells in the low micromolar range. Six HDAC inhibitors including suberoylanilide hydroxamine (SAHA), HC toxin, apicidin, panobinostat, SB939, and LAQ824 were additionally found to be highly effective HDP inducers in a porcine 3D4/31 macrophage cell line. Besides PBD3, other HDP genes such as PBD2 and cathelicidins (PG1-5) were concentration-dependently induced by those compounds in both IPEC-J2 and 3D4/31 cells. Furthermore, the antibacterial activities of 3D4/31 cells were augmented following 24 h exposure to HDAC inhibitors. In conclusion, a cell-based high-throughput screening assay was developed for the discovery of porcine HDP inducers, and newly identified HDP-inducing compounds may have potential to be developed as alternatives to antibiotics for applications in swine and possibly other animal species.


Assuntos
Peptídeos Catiônicos Antimicrobianos/farmacologia , Enterócitos/citologia , Ensaios de Triagem em Larga Escala/métodos , Infecção/veterinária , Doenças dos Suínos/tratamento farmacológico , Suínos/imunologia , beta-Defensinas/genética , Animais , Linhagem Celular , Epigênese Genética , Regulação da Expressão Gênica , Histona Desacetilases/genética , Histona Desacetilases/metabolismo , Infecção/tratamento farmacológico
18.
Int J Biol Macromol ; 118(Pt A): 610-616, 2018 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-29959006

RESUMO

BACKGROUND: It is assumed that genetic factors play crucial role in nephrolithiasis. The present study was conducted to explore the role of Human Transcription Factor-7 like-2 (TCF7L2) ß-defensin (DEFB1) and CD14 gene polymorphism in development and progression of nephrolithiasis. METHODS: The genotypes of TCF7L2, DEFB1 and CD14 polymorphism were determined in 240 nephrolithiasis patients and 240 healthy controls by restriction digestion method of PCR. The levels of serum TCF7L2, DEFB1, CD14, uric acid and other biochemical parameters were measured both in nephrolithiasis patients and healthy control. RESULTS: The patients and control groups showed 30% and 50% 1654 AA DEFB1 genotype respectively. The Allele frequency in case of patient's group was 63.67% while in control group it was 36.33%. The mean serum DEFB1 levels of the patients and control groups attained were 115.66 and 239.43 pg/mL respectively. The allele frequency of TCF7L2 in patients and controls were 44.17% and 70.0% for C-allele, 55.83% and 30.00% for T-allele respectively. The mean of serum TCF7L2 levels were significantly decreased in patients compared to control group. CONCLUSIONS: The present findings are first of its class that validates a considerable connection of DEFB1 and TCF7L2 gene polymorphisms with nephrolithiasis and could probably act as indicators to estimate the risk associated to nephrolithiasis.


Assuntos
Receptores de Lipopolissacarídeos/genética , Nefrolitíase/genética , Polimorfismo de Nucleotídeo Único , Proteína 2 Semelhante ao Fator 7 de Transcrição/genética , beta-Defensinas/genética , Feminino , Frequência do Gene , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade
19.
Int J Mol Sci ; 19(6)2018 Jun 02.
Artigo em Inglês | MEDLINE | ID: mdl-29865262

RESUMO

Alternative therapies are needed to reduce the use of antibiotics and incidence of drug-resistant Salmonellosis. Previous studies have revealed important roles of statins in regulating innate immunity. Therefore, we investigated the effects of statins on innate immunity in Salmonella-infected intestinal epithelial cells (IECs), which are involved in mucosal innate immunity. SW480 cells and Akt siRNA- or vitamin D receptor (VDR) siRNA-transfected SW480 cells were infected by wild-type S. Typhimurium strain SL1344 in the presence or absence of statins. The mRNA or protein expression was analyzed by real-time quantitative PCR or western blot analysis, respectively. Simvastatin or fluvastatin caused IL-8 (interleukin-8) suppression, but increased hBD-2 mRNA expression in Salmonella-infected SW480 cells. Both statins enhanced phosphorylated Akt and VDR expressions. Akt or VDR knockdown by siRNA counteracted the suppressive effect of simvastatin on IL-8 expression, whereas VDR knockdown diminished the enhanced hBD-2 expression in Salmonella-infected SW480 cells. Therefore, we observed differential regulation of statins on inflammatory IL-8 and anti-microbial hBD-2 expressions in Salmonella-infected IECs via PI3K/Akt signaling and VDR protein expression, respectively. The enhanced activity of antimicrobial peptides by statins in Salmonella-infected IECs could protect the host against infection, and modulation of pro-inflammatory responses could prevent the detrimental effects of overwhelming inflammation in the host.


Assuntos
Ácidos Graxos Monoinsaturados/farmacologia , Inibidores de Hidroximetilglutaril-CoA Redutases/farmacologia , Indóis/farmacologia , Interleucina-8/metabolismo , Mucosa Intestinal/microbiologia , Salmonella typhimurium , Sinvastatina/farmacologia , beta-Defensinas/metabolismo , Células CACO-2 , Linhagem Celular Tumoral , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Células Epiteliais/microbiologia , Fluvastatina , Regulação da Expressão Gênica , Humanos , Interleucina-8/genética , Mucosa Intestinal/metabolismo , beta-Defensinas/genética
20.
Int J Mol Sci ; 19(5)2018 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-29738484

RESUMO

Human abdominal subcutaneous adipose tissue consists of two individual layers—the superficial adipose tissue (SAT) and deep adipose tissue (DAT)—separated by the Scarpa’s fascia. The present study focuses on the analysis of morphological and immunological differences of primary adipocytes, adipose-derived stem cells (ASC), and tissue-infiltrating immune cells found in SAT and DAT. Adipocytes and stromal vascular fraction (SVF) cells were isolated from human SAT and DAT specimens and phenotypically characterized by in vitro assays. Ex vivo analysis of infiltrating immune cells was performed by flow cytometry. Primary adipocytes from SAT are larger in size but did not significantly differ in cytokine levels of LEPTIN, ADIPOQ, RBP4, CHEMERIN, DEFB1, VISFATIN, MCP1, or MSCF. ASC isolated from SAT proliferated faster and exhibited a higher differentiation potential than those isolated from DAT. Flow cytometry analysis indicated no specific differences in the relative numbers of ASC, epithelial progenitor cells (EPC), or CD3⁺ T-cells, but showed higher numbers of tissue-infiltrating macrophages in SAT compared to DAT. Our findings suggest that ASC isolated from SAT have a higher regenerative potential than DAT-ASC. Moreover, spatial proximity to skin microbiota might promote macrophage infiltration in SAT.


Assuntos
Obesidade/genética , Células-Tronco/metabolismo , Gordura Subcutânea Abdominal/metabolismo , Gordura Subcutânea/metabolismo , Adipócitos/metabolismo , Adipócitos/patologia , Adiponectina/genética , Adiponectina/metabolismo , Diferenciação Celular/genética , Proliferação de Células/genética , Células Cultivadas , Quimiocinas/genética , Quimiocinas/metabolismo , Citocinas/genética , Citocinas/metabolismo , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Leptina/genética , Leptina/metabolismo , Macrófagos/metabolismo , Macrófagos/patologia , Nicotinamida Fosforribosiltransferase/genética , Nicotinamida Fosforribosiltransferase/metabolismo , Obesidade/metabolismo , Obesidade/patologia , Proteínas Plasmáticas de Ligação ao Retinol/genética , Proteínas Plasmáticas de Ligação ao Retinol/metabolismo , Células-Tronco/patologia , Gordura Subcutânea Abdominal/patologia , beta-Defensinas/genética , beta-Defensinas/metabolismo
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