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1.
J Agric Food Chem ; 68(1): 138-146, 2020 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-31873011

RESUMO

Chronic exposure to minute doses of endotoxin elicits intestinal inflammation and impairs the gut barrier function, potentially resulting in systemic inflammation with elevated concentrations of biomarkers associated with metabolic syndrome. This study aimed to investigate the preventive effects of the Rubus suavissimus S. Lee leaf extract in a model of low-grade systemic inflammation. The predominant compounds found in the leaf extract are gallic acids, ellagic acid, and rubusoside. Results of the present study showed that R. suavissimus leaf extract supplementation could help preserve intestinal barrier integrity by upregulating the expression of the tight junction proteins [e.g., zonula occluden-1 (ZO-1) and junctional adhesion molecule-1 (JAMA)] and mucin (MUC)-4 and also suppress the release of plasmatic proinflammatory cytokines, including tumor necrosis factor (TNF)-α, interleukin (IL)-6, and monocyte chemotactic protein (MCP)-1, while restoring the production of anti-inflammatory adiponectin. We subsequently determined that the leaf extract contributes to restoring glucose metabolic homeostasis through maintaining insulin sensitivity. Furthermore, our mechanistic finding demonstrated that the R. suavissimus leaf extract supplementation prevented systemic inflammation-driven impaired insulin sensitivity in white adipose tissues (WATs) by modulating the expression of peroxisome-proliferator-activated receptor-γ (PPAR-γ) and insulin receptor subset-1 (IRS-1). Altogether, our findings suggest that the above supplementation contributes to restoring immune and metabolic homeostasis to enhance the overall health of the host thereby preventing the early onset of metabolic disorders such as obesity and type 2 diabetes.


Assuntos
Diabetes Mellitus Tipo 2/tratamento farmacológico , Inflamação/tratamento farmacológico , Rubus/química , Animais , Citocinas/genética , Citocinas/metabolismo , Diabetes Mellitus Tipo 2/genética , Diabetes Mellitus Tipo 2/metabolismo , Modelos Animais de Doenças , Feminino , Humanos , Inflamação/induzido quimicamente , Inflamação/genética , Inflamação/imunologia , Lipopolissacarídeos/efeitos adversos , Camundongos , Camundongos Endogâmicos C57BL , PPAR gama/genética , PPAR gama/metabolismo , Folhas de Planta/química , Proteínas de Junções Íntimas/genética , Proteínas de Junções Íntimas/metabolismo
2.
Biosci Biotechnol Biochem ; 84(1): 171-177, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31476130

RESUMO

We tested the hypothesis that α-lactalbumin inhibits the disruption of intestinal barrier function and liver cirrhosis by restoring gut-liver axis function in thioacetamide (TAA) -treated rats. Rat diets were supplemented with α-lactalbumin replacing 50% of dietary protein. After consuming α-lactalbumin for one week, rats were intraperitoneally injected with TAA twice a week for 14 weeks. The α-lactalbumin-enriched diet significantly inhibited the elevation of plasma alanine aminotransferase, aspartate aminotransferase, and hyaluronic acids. The supplement significantly reduced plasma lipopolysaccharide levels and increased occludin mRNA level. Hepatic fibrosis and regenerative nodules was developed and intestinal villi were shortened by TAA; α-Lactalbumin attenuated these histopathological changes. These results indicated that α-lactalbumin improved intestinal barrier function, suppressing endotoxin levels. These data also suggested that α-lactalbumin ameliorated the impairment of the gut-liver axis by TAA, inhibiting the development of liver cirrhosis.


Assuntos
Suplementos Nutricionais , Trato Gastrointestinal/efeitos dos fármacos , Lactalbumina/uso terapêutico , Cirrose Hepática Experimental/induzido quimicamente , Cirrose Hepática Experimental/dietoterapia , Fígado/efeitos dos fármacos , Substâncias Protetoras/uso terapêutico , Tioacetamida/farmacologia , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Fibrose/tratamento farmacológico , Trato Gastrointestinal/metabolismo , Expressão Gênica/efeitos dos fármacos , Ácido Hialurônico/sangue , Injeções Intraperitoneais , Lipopolissacarídeos/sangue , Fígado/metabolismo , Fígado/patologia , Cirrose Hepática Experimental/prevenção & controle , Masculino , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Tioacetamida/administração & dosagem , Proteínas de Junções Íntimas/genética
3.
APMIS ; 128(1): 10-19, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31642122

RESUMO

Atherogenesis is associated with chronic gut infections; however, the mechanisms are not clear. The aim of the study was to determine whether lipopolysaccharide of E. coli (E. coli LPS) may affect endothelial barrier and modify IL-10 expression in dendritic cells (DCs). Human umbilical vein endothelial cells (HUVECs) and monocyte-derived DCs were treated with E. coli LPS, apolipoprotein B100 (ApoB100) and 7-ketocholesterol (7-kCH) - harmful oxidized form of cholesterol. The effect of E. coli LPS, 7-kCH and ApoB100 on the barrier functions of HUVECs in real-time cell electric impedance sensing system (RTCA-DP) was assessed. Furthermore, the effect of 7-kCH and ApoB100 on barrier functions of HUVECs co-cultured with DCs previously treated with LPS was analyzed. Both E. coli LPS and 7-kCH decreased barrier functions of HUVECs and reduced tight junction protein mRNA expression, whereas ApoB100 increased endothelial barrier. In DCs, ApoB100 and E. coli LPS decreased IL-10 mRNA expression. In HUVECs co-cultured with DCs treated with LPS and subsequently pulsed with ApoB100 or 7-kCH, IL-10 mRNA expression was lower. E. coli LPS-exposed DCs diminished the protective effect of ApoB100 on endothelial integrity and led to the decrease in occludin mRNA expression. LPS potentially derived from gut microflora may destabilize endothelial barrier together with oxidized cholesterol and intensify the immunogenicity of ApoB100.


Assuntos
Células Dendríticas/efeitos dos fármacos , Escherichia coli/imunologia , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Interleucina-10/genética , Lipopolissacarídeos/imunologia , Junções Íntimas/efeitos dos fármacos , Apolipoproteína B-100/farmacologia , Células Cultivadas , Técnicas de Cocultura , Células Dendríticas/imunologia , Escherichia coli/química , Humanos , Cetocolesteróis/farmacologia , Lipopolissacarídeos/farmacologia , Ocludina/genética , Transdução de Sinais/efeitos dos fármacos , Proteínas de Junções Íntimas/genética
4.
J Biochem Mol Toxicol ; 33(11): e22397, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31557363

RESUMO

Fumonisins (Fums) are mycotoxins widely distributed in crops and feed, and ingestion of Fums-contaminated crops is harmful to animal health. The purpose of this study is to explore the effect of Fum B1 (FB1 ) on barrier functions of porcine intestinal epithelial cells, IPEC-J2, to clarify the intestinal toxicity of Fums in pigs. The results showed that the persistent treatment of FB1 significantly decreased the viability of IPEC-J2. Moreover, the expressions of Claudin 1, Occludin, Zonula Occluden-1 (ZO-1) on the messenger RNA (mRNA), and protein levels and MUC1 on the mRNA level were significantly inhibited after FB1 treatment, while the mRNA relative expression level of MUC2 was clearly increased. FB1 also enhanced the monolayer cell permeability of IPEC-J2. Importantly, FB1 promoted the expression of phosphorylated extracellular regulated protein kinase (p-ERK1/2 ). These data suggest that long-term treatment of FB1 can suppress IPEC-J2 proliferation, damage tight junctions of IPEC-J2, and regulate expression of mucins to induce the damage of barrier functions of porcine intestinal epithelial cells, which may be associated with the ERK1/2 phosphorylation pathway.


Assuntos
Células Epiteliais/metabolismo , Fumonisinas/farmacologia , Mucosa Intestinal/citologia , Micotoxinas/farmacologia , Permeabilidade/efeitos dos fármacos , Junções Íntimas/efeitos dos fármacos , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Fusarium/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Mucina-1/genética , Mucina-1/metabolismo , Mucina-2/genética , Mucina-2/metabolismo , Fosforilação/efeitos dos fármacos , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais/efeitos dos fármacos , Suínos , Proteínas de Junções Íntimas/genética , Proteínas de Junções Íntimas/metabolismo , Junções Íntimas/metabolismo
5.
Pol J Vet Sci ; 22(2): 345-353, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31269354

RESUMO

Tight junction proteins are important for the maintenance and repair of the intestinal mucosal barrier. The present study investigated relationships among tight junction protein gene expression, porcine epidemic diarrhea virus (PEDV) infection, and intestinal mucosal morphology in piglets. We compared the expression of six tight junction proteins (ZO-1, ZO-2, Occludin, Claudin-1, Claudin-4, and Claudin-5) between seven-day-old piglets infected with PEDV and normal piglets, as well as in PEDV-infected porcine intestinal epithelial cells (IPEC-J2). We also evaluated differences in mucosal morphology between PEDV-infected and normal piglets. The expression of six tight junction protein genes was lower in PEDV-infected piglets than in the normal animals. The expression of ZO-1, ZO-2, Occludin, and Claudin-4 in the intestine tissue was significantly lower (p⟨0.05) in PEDV-infected than in normal piglets. The expression of Claudin-5 in the jejunum was significantly lower in PEDV-infected piglets than in the normal animals (p⟨0.01). The expression of Claudin-1 and Claudin-5 genes in the ileum was significantly higher in PEDV-infected piglets than in normal piglets (p⟨0.01). Morphologically, the intestinal mucosa in PEDV-infected piglets exhibited clear pathological changes, including breakage and shedding of intestinal villi. In PEDV-infected IPEC-J2 cells, the mRNA expression of the six tight junction proteins showed a downward trend; in particular, the expression of the Occludin and Claudin-4 genes was significantly lower (p⟨0.01). These data suggest that the expression of these six tight junction proteins, especially Occludin and Claudin-4, plays an important role in maintaining the integrity of the intestinal mucosal barrier and resistance to PEDV infection in piglets.


Assuntos
Infecções por Coronavirus/veterinária , Vírus da Diarreia Epidêmica Suína , Doenças dos Suínos/virologia , Proteínas de Junções Íntimas/metabolismo , Animais , Linhagem Celular , Infecções por Coronavirus/virologia , Regulação da Expressão Gênica , Mucosa Intestinal/patologia , Suínos , Doenças dos Suínos/patologia , Proteínas de Junções Íntimas/genética
6.
Mol Med Rep ; 20(1): 393-400, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31115553

RESUMO

Various genomic and epigenetic modifications that occur during the development of cancer act as potential biomarkers for early diagnosis and treatment. Previous studies have demonstrated abnormal expression of the claudin (CLDN) tight junction (TJ) proteins in numerous types of human cancer. Reverse transcription­quantitative polymerase chain reaction and western blotting were employed to investigate variations in the expression of the CLDN TJ proteins in laryngeal non­neoplastic tissues and laryngeal squamous carcinoma tissues. It was revealed that CLDN2, CLDN4, CLDN5, CLDN6, CLDN9, CLDN11 and CLDN12 were undetectable in laryngeal squamous carcinoma tissues and laryngeal non­neoplastic tissues. Additionally, CLDN10 was expressed in laryngeal squamous carcinoma tissues and laryngeal non­neoplastic tissues; however, no significant difference was reported. Conversely, the expression levels of CLDN1 and CLDN7 mRNA and protein were downregulated in laryngeal squamous carcinoma tissues compared with in adjacent non­neoplastic tissues, whereas those of CLDN3 and CLDN8 were upregulated. A total of 80 samples of laryngeal squamous carcinoma and non­neoplastic tissues were analyzed for the expression of CLDN1, ­3, ­7 and ­8 via streptavidin­peroxidase immunohistochemical staining. It was revealed that the expression levels of CLDN1 and CLDN7 were downregulated in laryngeal squamous carcinoma tissues compared with in non­neoplastic mucosal tissues, whereas those of CLDN3 and CLDN8 were upregulated. Furthermore, the associations between CLDN expression and the clinicopathological factors of patients were analyzed. The expression levels of CLDN3 and CLDN7 were reported to be associated with distant metastasis and serve as potential predictors of poor prognosis. In conclusion, the findings of the present study demonstrated that the expression levels of CLDN1, ­3, ­7 and ­8 varied between laryngeal squamous carcinoma tissues and non­neoplastic tissues. The expression levels of these CLDNs may be useful molecular markers for the diagnosis of laryngeal carcinoma, and determining the metastasis and prognosis of this disease.


Assuntos
Claudina-1/genética , Claudina-3/genética , Claudinas/genética , Neoplasias Laríngeas/genética , Idoso , Biomarcadores Tumorais/genética , Carcinoma/genética , Carcinoma/patologia , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Neoplasias Laríngeas/patologia , Masculino , Pessoa de Meia-Idade , Prognóstico , RNA Mensageiro/genética , Proteínas de Junções Íntimas/genética
7.
Artigo em Inglês | MEDLINE | ID: mdl-31128282

RESUMO

Exposure to crude oil, its components, and oil dispersants during a major crude oil spill, such as the Deepwater Horizon Oil Spill, can elicit behavioral changes in animals and humans. However, the underlying mechanisms by which oil spill-related compounds alters behavior remains largely unknown. A major cause of behavioral changes generally is dysfunction of the blood-brain barrier (BBB). We investigated the impact of a crude oil high energy water accommodated fraction (HEWAF), benzo[a] pyrene (BaP; a major component of crude oil), and the oil dispersant COREXIT, on BBB function. BBB function was assessed by measuring transendothelial electrical resistance (TEER) of mouse brain microvascular endothelial cells (BMECs). Within 3 h after treatment, TEER was significantly reduced by exposure to high concentrations of all test compounds. TEER remained reduced in response to COREXIT after 48 h, but this effect waned in BMECs treated with HEWAF and BaP, with low-mid range concentrations inducing increased TEER compared to vehicle controls. At 48 h of treatment, BMEC viability was significantly reduced in response to 2% HEWAF, but was increased in response to BaP (25 and 50 µM). BMEC viability was increased with 80 ppm COREXIT, but was reduced with 160 ppm. Gene expression of tight junction-associated proteins (claudin-5 and tight junction protein-1), and cell adhesion receptor (vascular cell adhesion molecule-1) was reduced in response to HEWAF and COREXIT, but not BaP. Taken together, these data suggest that oil spill-related compounds markedly affect BBB function, and that these changes may underlie the observed behavioral changes due to crude oil exposure.


Assuntos
Benzo(a)pireno/toxicidade , Barreira Hematoencefálica/efeitos dos fármacos , Petróleo/toxicidade , Tensoativos/toxicidade , Proteínas de Junções Íntimas/genética , Animais , Barreira Hematoencefálica/fisiologia , Encéfalo/irrigação sanguínea , Encéfalo/citologia , Sobrevivência Celular , Células Cultivadas , Impedância Elétrica , Células Endoteliais/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Camundongos Endogâmicos C57BL
8.
J Anim Physiol Anim Nutr (Berl) ; 103(4): 1174-1184, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30990939

RESUMO

Eleutheroside B (EB) is a phenylpropanoid glycoside with anti-inflammatory properties, neuroprotective abilities, immunomodulatory effects, antinociceptive effects, and regulation of blood glucose. The aim of this study was to investigate the effects of EB on the barrier function in the intestinal porcine epithelial cells J2 (IPEC-J2). The IPEC-J2 cells were inoculated into 96-well plates at a density of 5 × 103 cells per well for 100% confluence. The cells were cultured in the presence of EB at concentrations of 0, 0.05, 0.10, and 0.20 mg/ml for 48 hr. Then, 0.10 mg/ml was selected as the suitable concentration for the estimation of transepithelial electric resistance (TEER) value, alkaline phosphatase activity, proinflammatory cytokines mRNA expression, tight junction mRNA and protein expression. The results of this study indicated that the supplementation of EB in IPEC-J2 cells decreased cellular membrane permeability and mRNA expression of proinflammatory cytokines, including interleukin-6 (IL-6), interferon-γ (INF-γ), and tumour necrosis factor-α (TNF-α). The supplementation of EB in IPEC-J2 cells increased tight junction protein expression and anti-inflammatory cytokines, interleukin 10 (IL-10) and transforming growth factor beta (TGF-ß). In addition, the western blotting and real-time quantitative polymerase chain reaction (RT-qPCR) results indicated that EB significantly (p < 0.05) increased the mRNA and protein expression of intestinal tight junction proteins, Claudin-3, Occludin, and Zonula Occludins protein-1 (ZO-1). Therefore, dietary supplementation of EB may increase intestinal barrier function, tight junction protein expression, anti-inflammatory cytokines, and decrease proinflammatory cytokines synthesis in IPEC-J2 cells.


Assuntos
Citocinas/metabolismo , Células Epiteliais/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Glucosídeos/farmacologia , Fenilpropionatos/farmacologia , Suínos , Proteínas de Junções Íntimas/metabolismo , Animais , Linhagem Celular , Proliferação de Células , Citocinas/genética , Relação Dose-Resposta a Droga , Glucosídeos/administração & dosagem , Jejuno/citologia , Fenilpropionatos/administração & dosagem , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas de Junções Íntimas/genética
9.
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
10.
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
11.
Toxicol In Vitro ; 58: 150-160, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30926360

RESUMO

Okadaic acid (OA) is a lipophilic phycotoxin that accumulates in the hepatopancreas and fatty tissue of shellfish. Consumption of highly OA-contaminated seafood leads to diarrhetic shellfish poisoning which provokes severe gastrointestinal symptoms associated with a disruption of the intestinal epithelium. Since the molecular mechanisms leading to intestinal barrier disruption are not fully elucidated, we investigated the influence of OA on intestinal tight junction proteins (TJPs) in differentiated Caco-2 cells. We found a concentration- and time-dependent deregulation of genes encoding for intestinal TJPs of the claudin family, occludin, as well as zonula occludens (ZO) 1 and 2. Immunofluorescence staining showed concentration-dependent effects on the structural organization of TJPs already after treatment with a subtoxic but human-relevant concentration of OA. In addition, changes in the structural organization of cytoskeletal F-actin as well as its associated protein ZO-1 were observed. In summary, we demonstrated effects of OA on TJPs in intestinal Caco-2 cells. TJP expressions were affected after treatment with food-relevant OA concentrations. These results might explain the high potential of OA to disrupt the intestinal barrier in vivo as its first target. Thereby the present data contribute to a better understanding of the OA-dependent induction of molecular effects within the intestinal epithelium.


Assuntos
Toxinas Marinhas/toxicidade , Ácido Okadáico/toxicidade , Proteínas de Junções Íntimas/metabolismo , Células CACO-2 , Humanos , Mucosa Intestinal/citologia , Mucosa Intestinal/metabolismo , Proteínas de Junções Íntimas/genética
12.
Food Funct ; 10(3): 1595-1608, 2019 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-30806428

RESUMO

B. longum has been reported to exert an alleviative effect on colitis, but the results also suggested significant differences among strains. Here in this study, we compared the effect of B. longum subsp. longum strains with different properties in EPS production on DSS-induced colitis. To investigate the alleviative effect of a ropy-exopolysaccharide (EPS) producing strain, Bifidobacterium longum subsp. longum YS108R, on experimental colitis, C57BL/6J mice (male, 6-8 weeks old) were randomly assigned to six groups (n = 8): normal control, DSS colitis and four DSS colitis groups orally administered with three B. longum subsp. longum strains (YS108R, C11A10B and HAN4-25) and B. animalis subsp. lactis BB12, respectively, in which YS108R produced ropy-EPS, C11A10B produced non-ropy-EPS, HAN4-25 did not produce EPS and BB12 was set as a positive control. Ropy-EPS producing strain YS108R could alleviate the symptoms and remit inflammation induced by DSS, in which YS108R could decrease the pro-inflammatory cytokine IL-6 and IL-17A levels after DSS challenge (from 102 ± 45.22 to 37.95 ± 20.33 pg mL-1 and from 22.14 ± 5.43 to 12.58 ± 2.74, p < 0.05), but another non-ropy-EPS producing strain C11A10B did not decrease the levels of these pro-inflammatory cytokines. Furthermore, YS108R could maintain the expression levels of genes related to the mucosal barrier, but strain HAN4-25, a non-EPS producer, was not able to maintain the expression levels of these genes after DSS challenge. Analysis of gut microbiota showed that DSS treatment significantly increased the relative abundance of Enterobacteriaceae and Peptostreptococcaceae (0.2623 ± 0.162 and 0.0512 ± 0.0361) and decreased the relative abundance of S24-7 (0.042 ± 0.0326); however, YS108R administration could decrease the relative abundance of Enterobacteriaceae and Peptostreptococcaceae to 0.0848 ± 0.0399 and 0.0032 ± 0.0047 and increase the relative abundance of S24-7 to 0.2625 ± 0.0566 (p < 0.05). The results showed that B. longum subsp. longum YS108R could alleviate DSS-induced colitis by modulating the inflammation related cytokines, maintenance of the normal mucosal barrier and reverting the change of microbiota.


Assuntos
Bifidobacterium longum/metabolismo , Colite/induzido quimicamente , Colite/tratamento farmacológico , Microbioma Gastrointestinal/efeitos dos fármacos , Mucosa Intestinal/efeitos dos fármacos , Polissacarídeos Bacterianos/farmacologia , Animais , Colo/efeitos dos fármacos , Colo/enzimologia , Colo/metabolismo , Citocinas/metabolismo , Sulfato de Dextrana/toxicidade , Regulação da Expressão Gênica/efeitos dos fármacos , Camundongos , Peroxidase/metabolismo , Polissacarídeos Bacterianos/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Proteínas de Junções Íntimas/genética , Proteínas de Junções Íntimas/metabolismo
13.
Food Funct ; 10(2): 1235-1242, 2019 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-30747184

RESUMO

Inflammation caused by either intrinsic or extrinsic toxins results in intestinal barrier dysfunction, contributing to inflammatory bowel disease (IBD) and other diseases. Vitamin A is a widely used food supplement although its mechanistic effect on intestinal structures is largely unknown. The goal of this study was to explore the mechanism by investigating the influence of vitamin A on the intestinal barrier function, represented by tight junctions. IPEC-J2 cells were differentiated on transwell inserts and used as a model of intestinal barrier permeability. Transepithelial electrical resistance (TEER) was used as an indicator of monolayer integrity and paracellular permeability. Western blot and the reverse transcriptase-polymerase chain reaction were used to assess the protein and mRNA expression of tight junction proteins. Immunofluorescence microscopy was used to evaluate the localization and expression of tight junctions. Differentiated cells were treated with a vehicle control (Ctrl), inflammatory stimulus (1 µg mL-1 LPS), LPS co-treatment with 0.1 µmol L-1 Vitamin A (1 µg mL-1 LPS + 0.1 µmol L-1 VA) and 0.1 µmol L-1 Vitamin A. LPS significantly decreased TEER by 24 hours, continuing this effect to 48 hours after application. Vitamin A alleviated the LPS-induced decrease of TEER from 12 hours to 48 hours, while Vitamin A alone enhanced TEER, indicating that Vitamin A attenuated LPS-induced intestinal epithelium permeability. Mechanistically, different concentrations of Vitamin A (0-20 µmol L-1) enhanced tight junction protein markers including Zo-1, Occludin and Claudin-1 both at protein and mRNA levels with an optimized dose of 0.1 µmol L-1. Immunofluorescence results demonstrated that majority of Zo-1 and Claudin-1 is located at the tight junctions, as we expected. LPS reduced the expression of these proteins and Vitamin A reversed LPS-reduced expression of these proteins, consistent with the results of western blot. In conclusion, Vitamin A improves the intestinal barrier function and reverses LPS-induced intestinal barrier damage via enhancing the expression of tight junction proteins.


Assuntos
Células Epiteliais/efeitos dos fármacos , Mucosa Intestinal/citologia , Lipopolissacarídeos/toxicidade , Proteínas de Junções Íntimas/metabolismo , Vitamina A/farmacologia , Animais , Linhagem Celular , Suínos , Proteínas de Junções Íntimas/genética
14.
Fish Physiol Biochem ; 45(2): 539-549, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30729411

RESUMO

Dietary arginine (Arg) could improve the intestinal structure and absorption of grass carp (Ctenopharyngodon idellus); however, the mechanism of Arg on intestinal morphology improvement was unclear. The present study aimed to explain the possible mechanism of the positive effect of Arg on intestinal epithelial cells of grass carp. An in vitro study was conducted through a primary culture model to assess the growth, cell viability, mRNA expressions of TOR signal pathway, and tight junction proteins of enterocytes after culture in the medium with 6 levels of Arg (0, 0.1, 0.2, 0.5, 1.0, and 2.0 mmol/L). The results showed that 0.5 mmol/L Arg improved the cell number and decreased the lactate dehydrogenase and creatine kinase activities in culture medium (P < 0.05). The alkaline phosphatase activity in cell lysis buffer was depressed by 1 and 2 mmol/L Arg (P < 0.05). The nitric oxide (NO) content showed an increasing trend with the Arg content (P < 0.05), whereas the NO synthase activity showed an opposite trend to NO. TOR expression was higher in 0.2 and 0.5 mmol/L groups, whereas S6K1 expression in 1.0 mmol/L and 2.0 mmol/L groups were lower (P < 0.05). The mRNA expressions of occludin, claudin 3, and claudin c in 0.5 mmol/L group were the highest, while ZO-1 and claudin b expressions were higher in 0.2 and 0.5 mmol/L groups (P < 0.05). This study indicated that Arg enhanced the growth and integrity of intestinal epithelial cells of grass carp through upregulation of mRNA expression of TOR signal pathway and tight junction proteins at an optimal Arg content of 0.2-0.5 mmol/L.


Assuntos
Arginina/farmacologia , Carpas/fisiologia , Enterócitos/efeitos dos fármacos , Serina-Treonina Quinases TOR/metabolismo , Proteínas de Junções Íntimas/metabolismo , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Arginina/administração & dosagem , Células Cultivadas , Dieta/veterinária , Relação Dose-Resposta a Droga , Enterócitos/fisiologia , Regulação da Expressão Gênica/efeitos dos fármacos , Transdução de Sinais , Proteínas de Junções Íntimas/genética
15.
Cell Mol Life Sci ; 76(10): 1987-2002, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30734065

RESUMO

At the blood-brain barrier (BBB), claudin (Cldn)-5 is thought to be the dominant tight junction (TJ) protein, with minor contributions from Cldn3 and -12, and occludin. However, the BBB appears ultrastructurally normal in Cldn5 knock-out mice, suggesting that further Cldns and/or TJ-associated marvel proteins (TAMPs) are involved. Microdissected human and murine brain capillaries, quickly frozen to recapitulate the in vivo situation, showed high transcript expression of Cldn5, -11, -12, and -25, and occludin, but also abundant levels of Cldn1 and -27 in man. Protein levels were quantified by a novel epitope dilution assay and confirmed the respective mRNA data. In contrast to the in vivo situation, Cldn5 dominates BBB expression in vitro, since all other TJ proteins are at comparably low levels or are not expressed. Cldn11 was highly abundant in vivo and contributed to paracellular tightness by homophilic oligomerization, but almost disappeared in vitro. Cldn25, also found at high levels, neither tightened the paracellular barrier nor interconnected opposing cells, but contributed to proper TJ strand morphology. Pathological conditions (in vivo ischemia and in vitro hypoxia) down-regulated Cldn1, -3, and -12, and occludin in cerebral capillaries, which was paralleled by up-regulation of Cldn5 after middle cerebral artery occlusion in rats. Cldn1 expression increased after Cldn5 knock-down. In conclusion, this complete Cldn/TAMP profile demonstrates the presence of up to a dozen TJ proteins in brain capillaries. Mouse and human share a similar and complex TJ profile in vivo, but this complexity is widely lost under in vitro conditions.


Assuntos
Barreira Hematoencefálica , Claudina-5/genética , Proteínas de Junções Íntimas/genética , Junções Íntimas/metabolismo , Adulto , Animais , Encéfalo/irrigação sanguínea , Encéfalo/metabolismo , Células Cultivadas , Claudina-5/metabolismo , Feminino , Expressão Gênica , Células HEK293 , Humanos , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Pessoa de Meia-Idade , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Proteínas de Junções Íntimas/metabolismo , Junções Íntimas/ultraestrutura
16.
Fish Shellfish Immunol ; 87: 546-558, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30716522

RESUMO

The present study was the first to investigate the effects of dietary vitamin A (VA) on the intestinal physical barrier function associated with oxidation, antioxidant system, apoptosis and cell-cellular tight junction (TJ) in the proximal (PI), mid (MI) and distal (DI) intestines of young grass carp (Ctenopharyngodon idella). Fish were fed graded levels of dietary VA for 10 weeks, and then a challenge test using an injection of Aeromonas hydrophila was conducted for 14 days. Results indicated that dietary VA deficiency caused oxidative damage to fish intestine partly by the reduced non-enzymatic antioxidant components glutathione (GSH) and VA contents as well as reduced antioxidant enzyme activities [not including manganese superoxide dismutase (MnSOD)]. Further results observed that the decreased antioxidant enzyme activities by VA deficiency were partly related to the down-regulation of their corresponding mRNA levels which were regulated by the down-regulation of NF-E2-related factor 2 (Nrf2) mRNA levels and up-regulation of kelch-like-ECH-associated protein (Keap1a) (rather than Keap1b) mRNA levels in three intestinal segments of fish. Meanwhile, VA deficiency up-regulated the mRNA levels of the apoptosis signalling [caspase-3, caspase-8, caspase-9 (rather than caspase-7)] associated with the inhibition of the target of rapamycin (TOR) signalling pathway in three intestinal segments of fish. Additionally, VA deficiency decreased the mRNA levels of TJ complexes [claudin-b, claudin-c, claudin-3, claudin-12, claudin-15a, occludin and zonula occludens-1 (ZO-1) in the PI, MI and DI, as well as claudin-7 and claudin-11a in the MI and DI] linked to the up-regulation of myosin light chain kinase (MLCK) signalling. These results suggested that VA deficiency impaired structural integrity in three intestinal segments of fish. Meanwhile, excessive VA also showed similar negative effects on these indexes. Taken together, the current study firstly demonstrated that VA deficiency impaired physical barrier functions associated with impaired antioxidant capacity, aggravated cell apoptosis and disrupted TJ complexes in the PI, MI and DI, but different segments performed different actions in fish. Based on protecting fish against protein oxidation, the optimal VA levels for grass carp were estimated to be 2622 IU/kg diet.


Assuntos
Carpas , Doenças dos Peixes/tratamento farmacológico , Intestinos/imunologia , Deficiência de Vitamina A/veterinária , Vitamina A/metabolismo , Vitaminas/metabolismo , Aeromonas hydrophila/fisiologia , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal/efeitos dos fármacos , Animais , Antioxidantes/metabolismo , Apoptose/efeitos dos fármacos , Dieta/veterinária , Suplementos Nutricionais/análise , Relação Dose-Resposta a Droga , Doenças dos Peixes/induzido quimicamente , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/patologia , Infecções por Bactérias Gram-Negativas/veterinária , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Intestinos/efeitos dos fármacos , Intestinos/patologia , Distribuição Aleatória , Proteínas de Junções Íntimas/genética , Vitamina A/administração & dosagem , Deficiência de Vitamina A/induzido quimicamente , Deficiência de Vitamina A/tratamento farmacológico , Vitaminas/administração & dosagem
17.
Poult Sci ; 98(5): 2211-2219, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30668786

RESUMO

Two dietary sources of zinc (ZnSO4 or organic Zn) were tested in chickens challenged with coccidiosis (Co) or coccidiosis plus Clostridium perfringens (CoCPF). On day 14, the chickens were orally gavaged with ∼5,000 Eimeria maxima sporulated oocysts. On day 19, 20, and 21 chickens challenged with C. perfringens were given a broth culture containing 108 cfu of this bacterium. Productive performance parameters were determined at d 14, 21, and 28. On day 21, necrotic enteritis (NE) lesions were scored, and intestinal permeability was evaluated. Jejunum and cecal tonsils were collected for morphology and gene expression analysis. On day 21, organic Zn improved BW gain by 18.6% (P = 0.07), and FCR by 12% (P = 0.09) in CoCPF challenged chickens vs. birds fed ZnSO4. From 1 to 28, organic Zn increased BW gain (P = 0.02), and improved FCR (P = 0.03) vs. birds fed ZnSO4. At 21 d, NE lesions were only observed in CoCPF birds (P < 0.001), and mortality due to NE was only observed when CoCPF birds were fed ZnSO4 (P = 0.001). Organic Zn fed birds had increased villus height in the jejunum (P = 0.005) and decreased intestinal permeability (P = 0.01) vs. ZnSO4. In the jejunum, organic Zn fed birds showed a downregulation of expression of IL-8 (P = 0.02), and upregulation of IL-10 (P = 0.05) in CoCPF birds vs. ZnSO4- CoCPF birds. As main effect, birds supplemented with organic Zn had higher mRNA expression of TLR-2 (P = 0.02) and IgA (P = 0.01). In the cecal tonsils, organic Zn fed birds showed upregulation of iNOS (P = 0.008) in CoCPF birds vs. ZnSO4-CoCPF birds. Organic Zn supplementation reduced intestinal permeability and attenuated intestinal inflammation of broilers co-challenged with coccidia and C. perfringens.


Assuntos
Ração Animal/análise , Galinhas , Infecções por Clostridium/veterinária , Coccidiose/veterinária , Doenças das Aves Domésticas/imunologia , Zinco/metabolismo , Animais , Proteínas Aviárias/genética , Proteínas Aviárias/metabolismo , Infecções por Clostridium/imunologia , Infecções por Clostridium/prevenção & controle , Clostridium perfringens/fisiologia , Coccidiose/imunologia , Coccidiose/prevenção & controle , Dieta/veterinária , Suplementos Nutricionais/análise , Eimeria/fisiologia , Expressão Gênica/efeitos dos fármacos , Inflamação/imunologia , Inflamação/prevenção & controle , Inflamação/veterinária , Intestinos/efeitos dos fármacos , Intestinos/fisiologia , Jejuno/anatomia & histologia , Jejuno/efeitos dos fármacos , Masculino , Doenças das Aves Domésticas/prevenção & controle , Distribuição Aleatória , Proteínas de Junções Íntimas/genética , Proteínas de Junções Íntimas/metabolismo
18.
Environ Toxicol ; 34(4): 548-555, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30698896

RESUMO

OBJECTIVE: To investigate the effect of paraquat (PQ) exposure on gene expression in neural stem cells as well as structures and functions of vascular endothelial cells. METHODS: RNA-Seq was used to explore the differentially expressed genes in human umbilical cord blood-neural stem cells (HUCB-NSCs) at different stages (eg, proliferation, early and late differentiation) in the presence of PQ. The effects of PQ on human umbilical vein endothelial cells (HUVECs), including cell proliferation, apoptosis, cytokines secretion, and expression of tight junction proteins, were assessed with CCK-8, flow cytometry, ELISA, and western blot analysis, individually. RESULTS: A total of 53 genes were up-regulated and 61 genes were down-regulated in PQ treated HUCB-NSCs, including seven genes associated with the differentiation of neural stem cells, for example, Gfap, S100B, Oct4, Gdf3, Sox1, Pax6, and Ngn1. PQ treatment significantly reduced the proliferation of HUVECs, inhibited cytokines secretion (VEGF, BFGF) and expressions of tight junction-associated protein (Claudin 1, Occludin, ZO-1), as well as induced significant apoptosis. CONCLUSION: Our study suggests that PQ impairs the development of nervous system by regulating the expression of genes associated with neural stem cell differentiation, as well as the structure and function of vascular endothelial cells, which together lead to abnormality in the nervous system.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células-Tronco Neurais/efeitos dos fármacos , Paraquat/toxicidade , Apoptose/efeitos dos fármacos , Apoptose/genética , Diferenciação Celular/genética , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Regulação para Baixo , Sequenciamento de Nucleotídeos em Larga Escala , Células Endoteliais da Veia Umbilical Humana/patologia , Humanos , Células-Tronco Neurais/patologia , Proteínas de Junções Íntimas/genética , Regulação para Cima
19.
FASEB J ; 33(4): 5312-5319, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30645152

RESUMO

Claudins (cldns) represent the largest family of transmembrane tight junction (TJ) proteins, determining organ-specific epithelial barrier properties. Because methods for the analysis of multiple cldn interaction are limited, we have established the heterologous Xenopus laevis oocyte expression system for TJ protein assembly and interaction analysis. Oocytes were injected with cRNA encoding human cldn-1, -2, or -3 or with a combination of these and were incubated in pairs for interaction analysis. Immunoblotting and immunohistochemistry were performed, and membrane contact areas were analyzed morphometrically and by freeze fracture electron microscopy. Cldns were specifically detected in membranes of expressing oocytes, and coincubation of oocytes resulted in adhesive contact areas that increased with incubation time. Adjacent membrane areas revealed specific cldn signals, including "kissing-point"-like structures representing homophilic trans-interactions of cldns. Contact areas of oocytes expressing a combination markedly exceeded those expressing single cldns, indicating effects on adhesion. Ultrastructural analysis revealed a self-assembly of TJ strands and a cldn-specific strand morphology.-Vitzthum, C., Stein, L., Brunner, N., Knittel, R., Fallier-Becker, P., Amasheh, S. Xenopus oocytes as a heterologous expression system for analysis of tight junction proteins.


Assuntos
Membrana Celular/metabolismo , Oócitos/metabolismo , Proteínas de Junções Íntimas/metabolismo , Animais , Claudina-1/genética , Claudina-1/metabolismo , Claudina-2/genética , Claudina-2/metabolismo , Claudina-3/genética , Claudina-3/metabolismo , Técnica de Fratura por Congelamento , Humanos , Immunoblotting , Imuno-Histoquímica , Microscopia Eletrônica , Ligação Proteica , Proteínas de Junções Íntimas/genética , Xenopus laevis
20.
Nutrients ; 11(1)2019 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-30621265

RESUMO

Gut dysbiosis and altered short-chain fatty acids are associated with ethanol-induced liver injury. SCFA are fermentation byproducts of the gut microbiota known to have many beneficial biological effects. We tested if a designer synbiotic could protect against ethanol-induced gut-liver injury. C57BL/6 female mice were exposed to chronic-binge ethanol feeding consisting of ethanol (5% vol/vol) for 10 days, followed by a single gavage (5 g/kg body weight) 6 h before euthanasia. A group of mice also received oral supplementation daily with a designer synbiotic, and another group received fecal slurry (FS); control animals received saline. Control mice were isocalorically substituted maltose dextran for ethanol over the entire exposure period. Ethanol exposure reduced expression of tight junction proteins in the proximal colon and induced hepatocyte injury and steatosis. Synbiotic supplementation not only mitigated losses in tight junction protein expression, but also prevented ethanol-induced steatosis and hepatocyte injury. Ethanol exposure also increased hepatic inflammation and oxidative stress, which was also attenuated by synbiotic supplementation. Mice receiving FS were not protected from ethanol-induced liver injury or steatosis. Results were associated with luminal SCFA levels and SCFA transporter expression in the proximal colon and liver. These results indicate supplementation with a designer synbiotic is effective in attenuating chronic-binge ethanol-induced gut-liver injury and steatosis in mice, and highlight the beneficial effects of the gut microbial fermentation byproducts.


Assuntos
Etanol/toxicidade , Enteropatias/induzido quimicamente , Enteropatias/prevenção & controle , Hepatopatias Alcoólicas/prevenção & controle , Simbióticos/administração & dosagem , Aldeídos/análise , Animais , Colo/química , Colo/metabolismo , Disbiose , Proteínas de Transporte de Ácido Graxo/análise , Ácidos Graxos Voláteis/análise , Ácidos Graxos Voláteis/metabolismo , Fígado Gorduroso Alcoólico/etiologia , Fezes , Feminino , Fermentação , Microbioma Gastrointestinal/fisiologia , Expressão Gênica/efeitos dos fármacos , Fígado/química , Camundongos , Camundongos Endogâmicos C57BL , Estresse Oxidativo/efeitos dos fármacos , Proteínas de Junções Íntimas/genética , Fator de Necrose Tumoral alfa/análise
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