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1.
Biol Res ; 53(1): 12, 2020 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-32209121

RESUMO

BACKGROUND: Mast cells (MCs) have been found to play a critical role during development of inflammatory bowel disease (IBD) that characterized by dysregulation of inflammation and impaired intestinal barrier function. However, the function of MCs in IBD remains to be fully elucidated. RESULTS: In our study, we used exosomes isolated from human mast cells-1 (HMCs-1) to culture with NCM460, HT-29 or CaCO2 of intestinal epithelial cells (IECs) to investigate the communication between MCs and IECs. We found that MCs-derived exosomes significantly increased intestinal epithelial permeability and destroyed intestinal barrier function, which is attributed to exosome-mediated functional miRNAs were transferred from HMCs-1 into IECs, leading to inhibit tight junction-related proteins expression, including tight junction proteins 1 (TJP1, ZO-1), Occludin (OCLN), Claudin 8 (CLDN8). Microarray and bioinformatic analysis have further revealed that a panel of miRNAs target different tight junction-related proteins. Interestingly, miR-223 is enriched in mast cell-derived exosome, which inhibit CLDN8 expression in IECs, while treatment with miR-223 inhibitor in HT-29 cells significantly reversed the inhibitory effect of HMCs-1-derived exosomes on CLDN 8 expression. Most importantly, enrichment of MCs accumulation in intestinal mucosa of patients with IBD compared with those healthy control. CONCLUSIONS: These results indicated that enrichment of exosomal miR-223 from HMCs-1 inhibited CLDN8 expression, leading to destroy intestinal barrier function. These finding provided a novel insight of MCs as a new target for therapeutic treatment of IBD.


Assuntos
Células Epiteliais/metabolismo , Mucosa Intestinal/metabolismo , Mastócitos/metabolismo , MicroRNAs/metabolismo , Animais , Células CACO-2/citologia , Bovinos , Células Cultivadas , Claudinas/metabolismo , Biologia Computacional , Exossomos/metabolismo , Humanos , Doenças Inflamatórias Intestinais/metabolismo , Ocludina/metabolismo , Permeabilidade , Análise Serial de Tecidos , Proteína da Zônula de Oclusão-1/metabolismo
2.
Chem Biol Interact ; 316: 108915, 2020 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-31816286

RESUMO

Stroke has been considered the second leading cause of death worldwide, and ischemic stroke accounts for the vast majority of stroke cases. Some of the main features of ischemic stroke are increased brain permeability, ischemia/reperfusion injury, oxidative stress, and acute inflammation. Antagonism of cysLT1R has been shown to provide cardiovascular and neural benefits. In the present study, we investigated the effects of the cysLT1R antagonist zafirlukast both in vivo and in vitro using a middle cerebral artery occlusion (MCAO) mouse model and human brain microvascular endothelial cells (HBMVECs). In vivo, we found that zafirlukast pretreatment could reduce MCAO-induced increased brain permeability by rescuing the expression levels of the tight junction proteins occludin and ZO-1. In vitro, we found that zafirlukast could suppress the increase in endothelial monolayer permeability induced by OGD/R via rescue of occludin and ZO-1 expression; additionally, we found that zafirlukast prevented OGD/R-induced degradation of the extracellular matrix via inhibition of MMP-2 and MMP-9 expression. Finally, we found that zafirlukast could also inhibit OGD/R-induced activation of the critical proinflammatory regulator NF-κB by preventing phosphorylation and nuclear translocation of p65 protein. Together, our findings demonstrate a promising role for zafirlukast in preventing damage induced by ischemic stroke and reperfusion injury.


Assuntos
Barreira Hematoencefálica/efeitos dos fármacos , Compostos de Tosil/farmacologia , Animais , Barreira Hematoencefálica/metabolismo , Infarto Encefálico/etiologia , Infarto Encefálico/prevenção & controle , Permeabilidade da Membrana Celular/efeitos dos fármacos , Modelos Animais de Doenças , Regulação para Baixo/efeitos dos fármacos , Células Endoteliais/citologia , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Infarto da Artéria Cerebral Média/complicações , Infarto da Artéria Cerebral Média/patologia , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 2 da Matriz/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , NF-kappa B/metabolismo , Ocludina/genética , Ocludina/metabolismo , Compostos de Tosil/uso terapêutico , Proteína da Zônula de Oclusão-1/genética , Proteína da Zônula de Oclusão-1/metabolismo
3.
J Agric Food Chem ; 68(1): 160-167, 2020 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-31825618

RESUMO

Inflammatory bowel disease (IBD) is a chronic inflammatory disease of intestinal mucosa and submucosa, characterized by the disruption of the intestinal epithelial barrier, increased production of inflammatory mediators, and excessive tissue injury. Intestinal epithelial cells, as well as microvascular endothelial cells, play important roles in IBD. To study the potential effects of kaempferol in IBD progress, we established a novel epithelial-endothelial cells coculture model to investigate the intestinal inflammation and barrier function. Data demonstrated an obvious increased transepithelial electrical resistance (TEER) (1222 ± 60.40 Ω cm2 vs 1371 ± 38.77 Ω cm2), decreased flux of FITC (180.8 ± 20.06 µg/mL vs 136.7 ± 14.78 µg/mL), and up-regulated occludin and claudin-2 expression in Caco-2 that was specifically cocultured with endothelial cells. Meanwhile, 80 µM kaempferol alleviated the drop of TEER, the increase of FITC flux, and the overexpression of interleukin-8 (IL-8) induced by 1 µg/mL lipopolysaccharide (LPS). Additionally, kaempferol also ameliorated the LPS-induced decrease of protein expression of zonula occludens-1 (ZO-1), occludin, and claudin-2, together with the inhibited protein expressions of the phosphorylation level of NF-κB and I-κB induced by LPS. Our results suggest that kaempferol alleviates the IL-8 secretion and barrier dysfunction of the Caco-2 monolayer in the LPS-induced epithelial-endothelial coculture model via inhibiting the NF-κB signaling pathway activation.


Assuntos
Células Endoteliais/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Mucosa Intestinal/citologia , Quempferóis/farmacologia , Lipopolissacarídeos/efeitos adversos , Células CACO-2 , Claudina-2/genética , Claudina-2/metabolismo , Técnicas de Cocultura , Células Endoteliais/citologia , Células Endoteliais/metabolismo , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Humanos , Inflamação/tratamento farmacológico , Inflamação/genética , Inflamação/metabolismo , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Lipopolissacarídeos/imunologia , Microvilosidades/efeitos dos fármacos , Microvilosidades/genética , Microvilosidades/metabolismo , Ocludina/genética , Ocludina/metabolismo , Proteína da Zônula de Oclusão-1/genética , Proteína da Zônula de Oclusão-1/metabolismo
4.
J Agric Food Chem ; 68(1): 168-175, 2020 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-31850758

RESUMO

Naringin is a polymethoxylated flavonoid commonly found in citrus species and has therapeutic potential in intestinal disorders. However, the effect and mechanism of naringin on gut-vascular barrier disruption has not yet been reported. This study aimed to investigate the distinguishing and selectively protective effects of naringin on tumor necrosis factor (TNF)-α-induced gut-vascular barrier disruption and elucidate the potential mechanism. In the present study, an in vitro gut-vascular barrier model composed of rat intestinal microvascular endothelial cells (RIMVECs) was studied. Evans blue-albumin efflux assay showed that naringin (50 µM) evidently protected the integrity of RIMVEC monolayer barriers against TNF-α-induced disruption. Naringin maintained the expression and distribution of tight junction proteins including zona occludin-1, occludin, claudin-1, and claudin-2. Additionally, naringin protected RIMVECs from TNF-α-induced apoptosis and cell migration suppression (41.1 ± 2.2 vs 51.1 ± 3.5%; 61.0 ± 5.1 vs 72.2 ± 6.2%). Our results indicate that naringin effectively ameliorates gut-vascular barrier disruption.


Assuntos
Células Endoteliais/efeitos dos fármacos , Flavanonas/farmacologia , Mucosa Intestinal/irrigação sanguínea , Mucosa Intestinal/efeitos dos fármacos , Extratos Vegetais/farmacologia , Substâncias Protetoras/farmacologia , Fator de Necrose Tumoral alfa/metabolismo , Animais , Apoptose/efeitos dos fármacos , Citrus/química , Claudina-1/genética , Claudina-1/metabolismo , Células Endoteliais/metabolismo , Técnicas In Vitro , Mucosa Intestinal/metabolismo , Microvasos/citologia , Microvasos/efeitos dos fármacos , Microvasos/metabolismo , Ocludina/genética , Ocludina/metabolismo , Ratos , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa/genética
5.
Cell Host Microbe ; 27(1): 25-40.e6, 2020 01 08.
Artigo em Inglês | MEDLINE | ID: mdl-31866426

RESUMO

Although a link between the gut microbiota and alcohol-related liver diseases (ALDs) has previously been suggested, the causative effects of specific taxa and their functions have not been fully investigated to date. Here, we analyze the gut microbiota of 410 fecal samples from 212 Korean twins by using the Alcohol Use Disorders Identification Test (AUDIT) scales to adjust for host genetics. This analysis revealed a strong association between low AUDIT scores and the abundance of the butyrate-producing genus Roseburia. When Roseburia spp. are administered to ALD murine models, both hepatic steatosis and inflammation significantly improve regardless of bacterial viability. Specifically, the flagellin of R. intestinalis, possibly through Toll-like receptor 5 (TLR5) recognition, recovers gut barrier integrity through upregulation of the tight junction protein Occludin and helps to restore the gut microbiota through elevated expression of IL-22 and REG3γ. Our study demonstrates that Roseburia spp. improve the gut ecosystem and prevent leaky gut, leading to ameliorated ALDs.


Assuntos
Clostridiales/metabolismo , Fígado Gorduroso Alcoólico/terapia , Microbioma Gastrointestinal , Adulto , Consumo de Bebidas Alcoólicas/efeitos adversos , Transtornos Relacionados ao Uso de Álcool/patologia , Animais , Clostridiales/isolamento & purificação , Disbiose/microbiologia , Fígado Gorduroso Alcoólico/metabolismo , Fezes/microbiologia , Feminino , Flagelina/metabolismo , Humanos , Mucosa Intestinal/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Pessoa de Meia-Idade , Ocludina/metabolismo
6.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 35(10): 892-896, 2019 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-31814565

RESUMO

Objective To investigate the effect of miR-129-5p on alcohol-induced intestinal epithelial permeability. Methods Real-time quantitative PCR was used to detect the expression level of miR-129-5p in sigmoid colon biopsies and alcohol-induced differentiated Caco-2 cells from patients with alcoholic liver disease and healthy people. Then, the differentiated Caco-2 cells were divided into 4 groups: the control group, the alcohol group (treatment with 50 mmol/L alcohol for 60 minutes), alcohol combined with miR-129-5p inhibitor group (transfection with miR-129-5p inhibitor before alcohol treatment), and alcohol combined with miR-129-5p inhibitor control group (transfection with miR-129-5p inhibitor control before alcohol treatment). After the alcohol treatment, the transepithelial electrical resistance (TER) was analyzed by a resistance meter, and the protein levels relevant to tight junction (occludin and ZO-1) were determined by Western blotting. Results The expression level of miR-129-5p was elevated in the biopsies of the patients with alcoholic liver disease compared with that in the healthy controls. Alcohol treatment increased miR-129-5p expression in the differentiated Caco-2 cells. Compared with the control group, TER and the protein levels of occludin and ZO-1 decreased in the alcohol group. Compared with the alcohol group, TER and the protein levels of occludin and ZO-1 were enhanced in the alcohol combined with miR-129-5p group; however, these changes were not altered in the alcohol combined with miR-129-5p inhibitor control group. Conclusion The miR-129-5p expression is elevated in sigmoid colon biopsies and alcohol-induced differentiated Caco-2 cells from patients with alcoholic liver disease. Inhibition of miR-129-5p can suppress alcohol-induced intestinal epithelial permeability.


Assuntos
Células Epiteliais/efeitos dos fármacos , Etanol/farmacologia , Mucosa Intestinal/citologia , MicroRNAs/genética , Células CACO-2 , Regulação para Baixo , Células Epiteliais/citologia , Humanos , Ocludina/metabolismo , Proteína da Zônula de Oclusão-1/metabolismo
7.
Med Sci Monit ; 25: 10045-10056, 2019 Dec 27.
Artigo em Inglês | MEDLINE | ID: mdl-31881016

RESUMO

BACKGROUND Barbaloin is one of the main medicinal ingredients of aloe vera, which displays various anti-inflammatory and anti-apoptosis properties in several inflammatory and fibrotic diseases. Our study evaluated its efficacy against dextran sulfate sodium (DSS)-induced colitis in rats. MATERIAL AND METHODS Ulcerative colitis (UC) rat models were established in vivo, and after barbaloin treatment, body weight and inflammation index were measured. Additionally, the signaling mechanism by which barbaloin protects against UC was investigated using LPS-infected Caco-2 cells. RESULTS Barbaloin could significantly reverse UC-induced weight loss and colon injury. Further, it could effectively increase the mRNA expression of IL-4 and IL-10 in colon tissues, while decreasing the expression of IFN-γ, IL-6, IL-1ß, and TNF-alpha. Furthermore, it significantly enhanced UC-inhibited atresia band 1 (ZO-1), occludin, and E-cadherin, and was also found to activate the AMPK signaling pathway. Additionally, si-RAN-induced knockdown, and overexpression assay showed that barbaloin could inhibit the UC-enhanced MLCK signaling pathway by activating the AMPK signaling pathway. CONCLUSIONS Barbaloin can effectively inhibit inflammation and reverse epithelial barrier function to protect against UC, possibly via activation of the AMPK signaling pathway.


Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Antracenos/uso terapêutico , Colite/tratamento farmacológico , Colite/enzimologia , Inflamação/patologia , Mucosa Intestinal/patologia , Transdução de Sinais , Animais , Antracenos/química , Antracenos/farmacologia , Células CACO-2 , Caderinas/metabolismo , Colite/patologia , Colite/fisiopatologia , Sulfato de Dextrana , Dextranos/sangue , Modelos Animais de Doenças , Fluoresceína-5-Isotiocianato/análogos & derivados , Humanos , Mediadores da Inflamação/metabolismo , Mucosa Intestinal/efeitos dos fármacos , Lipopolissacarídeos , Masculino , Quinase de Cadeia Leve de Miosina/metabolismo , Ocludina/metabolismo , Tamanho do Órgão/efeitos dos fármacos , Ratos Wistar , Transdução de Sinais/efeitos dos fármacos , Proteína da Zônula de Oclusão-1/metabolismo
8.
PLoS One ; 14(12): e0226780, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31887117

RESUMO

The pathogenesis of canine inflammatory bowel disease (IBD) involves complex interactions between mucosal immunity and the intestinal microbiota. Glucocorticoids are commonly administered to reduce mucosal inflammation and gastrointestinal signs. The study objective was to evaluate the effects of diet and oral prednisone on the spatial distribution of mucosal bacteria in IBD dogs. Eight dogs diagnosed with IBD were treated with immunosuppressive doses of prednisone. The mucosal microbiota from endoscopic biopsies of IBD dogs and healthy controls (HC; n = 15 dogs) was evaluated by fluorescence in situ hybridization (FISH) targeting the 16S rRNA genes of total bacteria and bacterial species relevant in canine/human IBD. Apicaljunction protein (AJP) expression using immunohistochemistry investigated the effect of medical therapy on intestinal barrier integrity. All IBD dogs had a reduction in GI signs following diet and prednisone therapy compared with baseline CIBDAI scores (P < 0.05). The mucosal microbiota of HC and diseased dogs was most abundant in free and adherent mucus. Only Lactobacilli were increased (P < 0.05) in the adherent mucus of IBD dogs compared to HC. The spatial distribution of mucosal bacteria was significantly different (P < 0.05) in IBD dogs following prednisone therapy, with higher numbers of Bifidobacteria and Streptococci detected across all mucosal compartments and increased numbers of Bifidobacterium spp., Faecalibacterium spp., and Streptococcus spp. present within adherent mucus. Differences in intestinal AJPs were detected with expression of occludin increased (P < 0.05) in IBD dogs versus HC. The expressions of occludin and E-cadherin were increased but zonulin decreased (P < 0.05 for each) in IBD dogs following prednisone therapy. In conclusion, the spatial distribution of mucosal bacteria differs between IBD and HC dogs, and in response to diet and glucocorticoid administration. Medical therapy was associated with beneficial changes in microbial community structure and enhanced mucosal epithelial AJP expression.


Assuntos
Dieta , Doenças do Cão , Glucocorticoides/uso terapêutico , Doenças Inflamatórias Intestinais/veterinária , Mucosa Intestinal/microbiologia , Microbiota , Animais , Bactérias/genética , Bactérias/isolamento & purificação , Caderinas/metabolismo , Demografia , Doenças do Cão/dietoterapia , Doenças do Cão/tratamento farmacológico , Doenças do Cão/microbiologia , Cães , Doenças Inflamatórias Intestinais/dietoterapia , Doenças Inflamatórias Intestinais/tratamento farmacológico , Doenças Inflamatórias Intestinais/microbiologia , Mucosa Intestinal/imunologia , Ocludina/metabolismo , Prednisona/uso terapêutico
9.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 35(9): 800-805, 2019 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-31750821

RESUMO

Objective To investigate the role and mechanism of histone deacetylase 3 (HDAC3) in alcohol-induced inflammation and permeability of intestinal epithelial cells. Methods To select the proper concentration of alcohol, differentiated Caco-2 cells were treated with different concentrations (10, 25, 50, 100 and 200 mmol/L) of alcohol, and then cell viability was assayed by MTT assay; the mRNA and protein levels of HDAC3 were analyzed by real-time PCR and Western blot analysis. Differentiated Caco-2 cells were divided into three groups: control group, alcohol group (treatment with 50 mmol/L alcohol for 60 minutes), and alcohol combined with HDAC3 inhibitor group (pretreatment with 2 µmol/L RGFP966 1 hour before alcohol). ELISA was performed to detect tumor necrosis factor α (TNF-α) level in cell supernatant. Transepithelial electrical resistance (TER) was measured using a resistance meter. Western blot analysis was used to determine the protein levels relevant to tight junction (occludin and claudin-1) and NF-κB activation (IκB and phosphorylated NF-κBp65). Results Alcohol at 10, 25 and 50 mmol/L did not affect cell viability. The mRNA and protein expression levels of HDAC3 increased in a dose-dependent manner after alcohol treatment at these concentration s. Compared with the control group, TNF-α and phosphorylated NF-κBp65 levels increased, whereas TER and protein levels of occludin, claudin-1 and IκB decreased in the alcohol group. Compared with the alcohol group, TNF-α and phosphorylated NF-κBp65 levels were reduced, while TER and protein levels of occludin, claudin-1 and IκB were elevated in the alcohol combined with HDAC3 inhibitor group. Conclusion HDAC3 inhibition can attenuate alcohol-induced inflammation and permeability of intestinal epithelial cells, which may be related to the inactivation of NF-κB.


Assuntos
Etanol/efeitos adversos , Inibidores de Histona Desacetilases/farmacologia , Mucosa Intestinal/efeitos dos fármacos , Fator de Transcrição RelA/antagonistas & inibidores , Células CACO-2 , Claudina-1/metabolismo , Histona Desacetilases , Humanos , Proteínas I-kappa B/metabolismo , Inflamação , Mucosa Intestinal/patologia , Ocludina/metabolismo , Fator de Transcrição RelA/metabolismo , Fator de Necrose Tumoral alfa
10.
Mol Carcinog ; 58(12): 2316-2326, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31553086

RESUMO

Primary tumor can induce the formation of premetastatic niche. The hyperpermeability of the vessels in the premetastatic niche is the first step in the development of metastasis. However, the cellular and molecular mechanisms of vascular hyperpermeability remain to be elucidated. In this study, 4T1 breast cells were injected into the breasts of mice to establish a tumor model. Our results showed that primary tumors induced hyperpermeability of the vessels in the premetastatic lung. Subsequent studies showed that the level of vascular endothelial growth factor (VEGF) was elevated in the tumor-bearing mice serum and the levels of tight junction (TJ) proteins occludin and ZO-1 were decreased in the premetastatic lung. In vitro studies demonstrated that VEGF increased the permeability of dextran and decreased the levels of occludin and ZO-1 in human umbilical vein endothelial cells. Moreover, the hyperpermeability of vessels and the degradation of occludin was blocked by bevacizumab. Overexpression of occludin alleviated the VEGF-induced hyperpermeability. Further investigations revealed that VEGF-induced occludin phosphorylation at Ser-490 and ubiquitination. Finally, we showed that VEGF accelerated the process of occludin degradation through the ubiquitin-proteasome system. In conclusion, primary tumor-secrete VEGF induce the occludin phosphorylation/ubiquitination and downregulation, resulting in the disruption of TJs and hyperpermeability of vessels in premetastatic lung. The occludin phosphorylation/ubiquitination pathway may be the mechanism of VEGF-induced vascular hyperpermeability in the lung premetastatic niche.


Assuntos
Permeabilidade Capilar/efeitos dos fármacos , Pulmão/metabolismo , Neoplasias Experimentais/metabolismo , Ocludina/metabolismo , Fator A de Crescimento do Endotélio Vascular/farmacologia , Animais , Linhagem Celular Tumoral , Células Cultivadas , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Pulmão/irrigação sanguínea , Pulmão/patologia , Camundongos Endogâmicos BALB C , Mutação de Sentido Incorreto , Neoplasias Experimentais/genética , Neoplasias Experimentais/patologia , Ocludina/genética , Fosforilação/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Junções Íntimas/efeitos dos fármacos , Junções Íntimas/metabolismo , Ubiquitinação/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/metabolismo
11.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 41(4): 506-511, 2019 Aug 30.
Artigo em Chinês | MEDLINE | ID: mdl-31484613

RESUMO

To investigate the expressions of mucosal barrier proteins in colon cell line DLD-1 under hypoxic environment in vitro and its mechanism. Methods After DLD-1 cells were treated separately with hypoxia(l% O2),vitamin D(100 nmol/L),or vitamin D plus hypoxia for 48 hours,the expressions of vitamin D receptor(VDR),tight junction proteins zonula occludens-1(ZO-1),occludin,Claudin-1,and adherent junction protein(E-cadherin)were determined by Western blot.Stable VDR knock-down(Sh-VDR)DLD-1 cell line and control DLD-1 cell line were established by lentivirus package technology and the protein expressions after hypoxia treatment were detected. Results Compared with control group,the expressions of occludin,Claudin-1,and VDR increased significantly after hypoxia treatment(all P<0.001).In addition to the protein expressions of occludin,Claudin-1 and VDR,the expressions of ZO-1 and E-cadherin were also obviously higher in vitamin D plus hypoxia group than in single vitamin D treatment group(all P<0.001).After hypoxia treatment,Sh-VDR cell line showed significantly decreased expressions of ZO-1(P<0.001),occludin(P<0.05),Claudin-1(P<0.01)and E-cadherin(P<0.001)when compared with untreated Sh-VDR cell line. Conclusion VDR acts as a regulator for the expressions of intestinal mucosal barrier proteins under hypoxia environment in DLD-1 colon cell line,indicating that VDR pathway may be another important protective mechanism for gut barrier in low-oxygen environment.


Assuntos
Colo/citologia , Receptores de Calcitriol/metabolismo , Antígenos CD/metabolismo , Caderinas/metabolismo , Hipóxia Celular , Linhagem Celular , Claudina-1/metabolismo , Humanos , Ocludina/metabolismo , Junções Íntimas , Vitamina D/farmacologia , Proteína da Zônula de Oclusão-1/metabolismo
12.
Invest Ophthalmol Vis Sci ; 60(12): 3842-3853, 2019 09 03.
Artigo em Inglês | MEDLINE | ID: mdl-31529081

RESUMO

Purpose: Outer blood retinal barrier breakdown is a neglected feature of diabetic retinopathy (DR). We demonstrated that the agonism of the δ opioid receptor (DOR) by epicatechin preserves the tight junction proteins in ARPE-19 cells under diabetic conditions. Presently, we aimed to evaluate the possible role of the DOR on the maintenance of tight junction of RPE layer and on the early markers of experimental DR. Methods: DR markers and external retinal tight junction proteins were evaluated in CL57B diabetic mice submitted to intravitreous injection of short hairpin RNA (shRNA)-DOR (108 transducing units [TU]/mL) treated or not with DOR agonist (0.05 g/animal/d of epicatechin in drinking water) for 16 weeks. The presence of DOR in human retina from postmortem eyes from diabetic and nondiabetic donors were also performed. Results: DOR is present in RPE layer and in neuro retina. The treatment with DOR agonist prevented the upregulation of the early markers of retinopathy (glial fibrillary acidic protein, VEGF) and the downregulation of pigment epithelium-derived factor, occludin, claudin-1, and zonula occludens-1 tight junction expressions. The silencing of DOR in retina of diabetic mice partially abolished the protective effects of epicatechin. In human retina specimens, DOR is present throughout the retina, similarly in nondiabetic and diabetic donors. Conclusions: This set of experiments strongly indicates that the DOR agonism preserves RPE tight junctions and reduces the early markers of retinopathy in model of diabetes. These novel findings designate DOR as a potential therapeutic tool to treat DR with preservation of the RPE tight junction proteins.


Assuntos
Catequina/farmacologia , Diabetes Mellitus Experimental/prevenção & controle , Retinopatia Diabética/prevenção & controle , Receptores Opioides delta/agonistas , Epitélio Pigmentado da Retina/metabolismo , Junções Íntimas/metabolismo , Idoso , Animais , Glicemia/metabolismo , Western Blotting , Claudina-1/metabolismo , Diabetes Mellitus Experimental/metabolismo , Retinopatia Diabética/metabolismo , Retinopatia Diabética/patologia , Impedância Elétrica , Proteínas do Olho/metabolismo , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Proteína Glial Fibrilar Ácida/metabolismo , Humanos , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Pessoa de Meia-Idade , Fatores de Crescimento Neural/metabolismo , Ocludina/metabolismo , RNA Interferente Pequeno , Reação em Cadeia da Polimerase em Tempo Real , Receptores Opioides delta/metabolismo , Serpinas/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Proteína da Zônula de Oclusão-1/metabolismo
13.
Chin J Nat Med ; 17(7): 498-505, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31514981

RESUMO

The aim of this study was to explore the neuroprotective effect and mechanism of XingNaoJing injections (XNJ) on cerebral ischemia injury and blood-brain barrier (BBB) disruption. Middle cerebral artery occlusion (MCAO) method was applicated to establish the model of cerebral ischemia/reperfusion (I/R) injury in rats. BBB permeability after I/R injury was assessed with the leaking amount of Evans Blue and the expression of occludin and ZO-1. The expression of NOD-like receptor family, pyrin domain containing (NLRP3) was checked to explore the inhibition of inflammation by XNJ. The results showed that XNJ could significantly increase the survival percent, decrease the infarct area and ameliorate neurological deficits and brain damage after I/R injury. Leaking amount of Evans Blue was reduced by XNJ, and the expression of tight junction protein, occludin and ZO-1 was also up-regulated by XNJ, which showed a role of protection on BBB disruption. The expression of NLRP3 was inhibited after exposure of XNJ, which was associated with inhibition of the inflammatory response. In summary, XNJ could suppress NLRP3 inflammasomes and improve BBB disruption and brain damage in rats after cerebral I/R injury, which provided a beneficial insight to further explore XNJ.


Assuntos
Barreira Hematoencefálica/efeitos dos fármacos , Isquemia Encefálica/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Inflamassomos/efeitos dos fármacos , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Fármacos Neuroprotetores/uso terapêutico , Traumatismo por Reperfusão/tratamento farmacológico , Animais , Barreira Hematoencefálica/metabolismo , Barreira Hematoencefálica/patologia , Isquemia Encefálica/metabolismo , Isquemia Encefálica/patologia , Permeabilidade Capilar , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/farmacologia , Masculino , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Fármacos Neuroprotetores/administração & dosagem , Fármacos Neuroprotetores/farmacologia , Ocludina/genética , Ocludina/metabolismo , Ratos Sprague-Dawley , Traumatismo por Reperfusão/metabolismo , Traumatismo por Reperfusão/patologia , Análise de Sobrevida , Regulação para Cima/efeitos dos fármacos , Proteína da Zônula de Oclusão-1/genética , Proteína da Zônula de Oclusão-1/metabolismo
14.
Food Funct ; 10(9): 5323-5332, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31389456

RESUMO

Oxidative stress is considered to play an important role in the cerebral ischemia-reperfusion injury. The nuclear transcription factor erythroid-2-related factor 2 (Nrf2)/NAD(P)H dehydrogenase [quinone] 1 (NQO1) pathway has been considered as a potential target for neuroprotection in cerebral ischemia-reperfusion injury. Nomilin (NOM) is a limonoid compound obtained from the extracts of citrus fruits. The purpose of our study was to determine whether NOM could exert beneficial effects in cerebral ischemia-reperfusion rats. Firstly, NOM treatment significantly mitigated cell death and decreased lactate dehydrogenase (LDH) release and ROS production in SH-SY5Y cells induced by oxygen-glucose deprivation (OGD), which was almost abolished by Nrf2 knockdown. Secondly, NOM improved infarct area, brain edema and neurological deficits in an experimental stroke rat model via middle cerebral artery occlusion (MCAO). Furthermore, NOM attenuated blood-brain barrier (BBB) disruption in MCAO rats, which might be associated with alleviating the loss of tight junction proteins, including ZO-1 and occludin-5. Further results revealed that NOM treatment effectively mitigated oxidative stress and facilitated the expressions of Nrf2 and NQO1, which might confirm that the loss of tight junction proteins in the microvasculature was likely mediated by oxidative stress. In conclusion, our study provided evidence that the protective effects of NOM in cerebral ischemia-reperfusion rats were related to the Nrf2/NQO1 pathway.


Assuntos
Benzoxepinas/administração & dosagem , Barreira Hematoencefálica/efeitos dos fármacos , Limoninas/administração & dosagem , Fator 2 Relacionado a NF-E2/metabolismo , Fármacos Neuroprotetores/administração & dosagem , Substâncias Protetoras/administração & dosagem , Traumatismo por Reperfusão/prevenção & controle , Animais , Barreira Hematoencefálica/metabolismo , Isquemia Encefálica/genética , Isquemia Encefálica/metabolismo , Isquemia Encefálica/cirurgia , Humanos , Masculino , NAD(P)H Desidrogenase (Quinona)/genética , NAD(P)H Desidrogenase (Quinona)/metabolismo , Fator 2 Relacionado a NF-E2/genética , Ocludina/genética , Ocludina/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Traumatismo por Reperfusão/genética , Traumatismo por Reperfusão/metabolismo , Proteína da Zônula de Oclusão-1/genética , Proteína da Zônula de Oclusão-1/metabolismo
15.
Gastroenterology ; 157(5): 1323-1337, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31401143

RESUMO

BACKGROUND & AIMS: Epithelial tight junctions are compromised in gastrointestinal disease. Processes that contribute to the resulting barrier loss include endocytic occludin removal from the tight junction and reduced occludin expression. Nevertheless, the relatively-normal basal phenotype of occludin knockout (KO) mice has been taken as evidence that occludin does not contribute to gastrointestinal barrier function. We asked whether stress could unmask occludin functions within intestinal epithelia. METHODS: Wildtype (WT), universal and intestinal epithelial-specific occludin KO, and villin-EGFP-occludin transgenic mice as well as WT and occludin knockdown (KD) Caco-2BBe cell monolayers were challenged with DSS, TNBS, staurosporine, 5-FU, or TNF. Occludin and caspase-3 expression were assessed in patient biopsies. RESULTS: Intestinal epithelial occludin loss limited severity of DSS- and TNBS-induced colitis due to epithelial resistance to apoptosis; activation of both intrinsic and extrinsic apoptotic pathways was blocked in occludin KO epithelia. Promoter analysis revealed that occludin enhances CASP3 transcription and, conversely, that occludin downregulation reduces caspase-3 expression. Analysis of biopsies from Crohn's disease and ulcerative colitis patients and normal controls demonstrated that disease-associated occludin downregulation was accompanied by and correlated with reduced caspase-3 expression. In vitro, cytokine-induced occludin downregulation resulted in reduced caspase-3 expression and resistance to intrinsic and extrinsic pathway apoptosis, demonstrating an overall protective effect of inflammation-induced occludin loss. CONCLUSIONS: The tight junction protein occludin regulates apoptosis by enhancing caspase-3 transcription. These data suggest that reduced epithelial caspase-3 expression downstream of occludin downregulation is a previously-unappreciated anti-apoptotic process that contributes to mucosal homeostasis in inflammatory conditions.


Assuntos
Apoptose , Caspase 3/metabolismo , Colite/enzimologia , Colo/enzimologia , Células Epiteliais/enzimologia , Mucosa Intestinal/enzimologia , Ocludina/metabolismo , Animais , Células CACO-2 , Estudos de Casos e Controles , Caspase 3/deficiência , Caspase 3/genética , Colite/induzido quimicamente , Colite/genética , Colite/patologia , Colite Ulcerativa/enzimologia , Colite Ulcerativa/patologia , Colo/patologia , Doença de Crohn/enzimologia , Doença de Crohn/patologia , Sulfato de Dextrana , Modelos Animais de Doenças , Células Epiteliais/patologia , Humanos , Mucosa Intestinal/patologia , Camundongos Endogâmicos C57BL , Camundongos Knockout , Ocludina/deficiência , Ocludina/genética , Transdução de Sinais , Ácido Trinitrobenzenossulfônico , Proteína da Zônula de Oclusão-1/genética , Proteína da Zônula de Oclusão-1/metabolismo
16.
Int J Mol Sci ; 20(16)2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31443263

RESUMO

Intestinal inflammation is an inflammatory disease resulting from immune dysregulation in the gut. It can increase the risk of enteric cancer, which is a common malignancy globally. As a new class of anti-inflammatory agents, native peptides have potential for use in the treatment of several intestinal inflammation conditions; however, their potential cytotoxicity and poor anti-inflammatory activity and stability have prevented their development. Hybridization has been proposed to overcome this problem. Thus, in this study, we designed a hybrid peptide (LL-37-TP5, LTP) by combing the active centre of LL-37 (13-36) with TP5. The half-life and cytotoxicity were tested in vitro, and the hybrid peptide showed a longer half-life and lower cytotoxicity than its parental peptides. We also detected the anti-inflammatory effects and mechanisms of LTP on Lipopolysaccharide (LPS)-induced intestinal inflammation in murine model. The results showed that LTP effectively prevented LPS-induced weight loss, impairment of intestinal tissues, leukocyte infiltration, and histological evidence of inflammation. Additionally, LTP decreased the levels of tumour necrosis factor-alpha, interferon-gamma, and interleukin-6; increased the expression of zonula occludens-1 and occludin; and reduced permeability in the jejunum of LPS-treated mice. Notably, LTP appeared to be more potent than the parental peptides LL-37 and TP5. The anti-inflammatory effects of LTP may be associated with the neutralization of LPS, inhibition of oxidative stress, and inhibition of the NF-κB signalling pathway. The findings of this study suggest that LTP might be an effective therapeutic agent for treating intestinal inflammation.


Assuntos
Antioxidantes/uso terapêutico , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Mucosa Intestinal/metabolismo , Lipopolissacarídeos/toxicidade , Animais , Interleucina-6/metabolismo , Mucosa Intestinal/efeitos dos fármacos , Camundongos , NF-kappa B/metabolismo , Ocludina/metabolismo , Células RAW 264.7 , Transdução de Sinais/efeitos dos fármacos , Proteína da Zônula de Oclusão-1/metabolismo
17.
Mol Med Rep ; 20(4): 3292-3300, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31432190

RESUMO

Previous studies have demonstrated the effects of hyperuricemia on the damage to target organs, including the kidneys, joints and the heart. However, it is unclear whether hyperuricemia results in damage to the intestines. The aim of the present study was to investigate intestinal barrier dysfunction in a mouse model of hyperuricemia constructed by knocking out the urate oxidase (Uox) gene. The morphology of the intestine was assessed via hematoxylin and eosin, and alcian blue staining. The serum and intestinal tissue levels of uric acid, tumor necrosis factor (TNF)­α and interleukin (IL)­6, in addition to the presence of uremic toxins in the serum, were assessed. The levels of diamine oxidase (DAO), D­lactate (D­LAC) and endotoxins in the serum, which are markers of the intestinal permeability, were measured using ELISA. The expression of the intestinal tight junction proteins zona occludens­1 (ZO­1) and occludin were detected by reverse transcription­quantitative polymerase chain reaction, western blotting and immunohistochemical analysis. The Uox­knockout mice spontaneously developed hyperuricemia. Histopathological analysis indicated notable intestinal defects including sparse villi, mucosal edema and a declining mucus layer in hyperuricemic mice. The expression levels of ZO­1 and occludin in the intestines were downregulated, and the serum levels of DAO, D­LAC and endotoxins were higher in the hyperuricemic mice, compared with control mice. The serum and intestinal tissue levels of IL­6 and TNF­α were significantly increased. Additionally, the expression levels of the serum uremic toxins, serum creatinine, blood urea nitrogen were significantly increased in hyperuricemic mice compared with the control mice, while only a marked increase in indoxyl sulfate (IS) and p­cresol sulfate was reported. Collectively, the results of the present study suggested that intestinal barrier dysfunction and subsequent enhanced intestinal permeability may occur as a result of hyperuricemia in mice. Furthermore, we proposed that the loss of intestinal epithelium barrier function may be associated with uric acid­induced inflammatory responses; however, further investigation is required.


Assuntos
Hiperuricemia , Mucosa Intestinal/metabolismo , Urato Oxidase/deficiência , Ácido Úrico/metabolismo , Animais , Modelos Animais de Doenças , Hiperuricemia/genética , Hiperuricemia/metabolismo , Hiperuricemia/patologia , Interleucina-6/genética , Interleucina-6/metabolismo , Mucosa Intestinal/patologia , Camundongos , Camundongos Knockout , Ocludina/genética , Ocludina/metabolismo , Permeabilidade , Junções Íntimas/genética , Junções Íntimas/metabolismo , Junções Íntimas/patologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Proteína da Zônula de Oclusão-1/genética , Proteína da Zônula de Oclusão-1/metabolismo
18.
Food Funct ; 10(7): 4350-4360, 2019 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-31276135

RESUMO

Cinnamon is known to have several physiological effects; the effects of Cinnamomum japonicum Sieb. on anti-inflammation and tight junctions were investigated in the cellular intestinal inflammation model. Cinnamon subcritical water extract (CSWE) significantly down-regulated the protein and expression levels of nitrite, prostaglandin E2 (PGE2), interleukin (IL)-6, IL-8, tumor necrosis factor (TNF)-α, nuclear factor kappa B (NF-κB) activity, and the phosphorylation of the factors of the NF-κB pathway. It also significantly decreased the permeability but increased the transepithelial electrical resistance (TEER) value and the protein and expression levels of tight junction proteins (i.e., zonula occludens (ZO)-1, occludin, and claudin-1). Furthermore, cinnamic acid and cinnamaldehyde, the major components of C. japonicum, inhibited the phosphorylation of the NF-κB pathway and increased the tight junction protein expression. CSWE from C. japonicum may improve intestinal health by enhancing tight junctions and inhibiting inflammation of the intestines.


Assuntos
Cinnamomum zeylanicum/química , Inflamação/tratamento farmacológico , Intestinos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Junções Íntimas/efeitos dos fármacos , Animais , Células CACO-2 , Claudina-1/metabolismo , Técnicas de Cocultura , Dinoprostona , Humanos , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Camundongos , NF-kappa B/metabolismo , Nitritos/metabolismo , Ocludina/metabolismo , Permeabilidade/efeitos dos fármacos , Fosforilação , Células RAW 264.7 , Proteínas de Junções Íntimas/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Água/química
19.
Int J Mol Sci ; 20(14)2019 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-31315208

RESUMO

Enterotoxigenic Escherichia coli (ETEC) infection destroys the intestinal barrier integrity, in turn, disrupting intestinal homoeostasis. Low-molecular-weight chitosan (LMWC) is a water-soluble chitosan derivative with versatile biological properties. Herein, we examined whether LMWC could relieve ETEC-induced intestinal barrier damage in weaned pigs. Twenty-four weaned pigs were allotted to three treatments: (1) non-infected control; (2) ETEC-infected control; and (3) ETEC infection + LMWC supplementation (100 mg/kg). On day 12, pigs in the infected groups were administered 100 mL of ETEC at 2.6 × 109 colony-forming units/mL to induce intestinal barrier injury. Three days later, serum samples were obtained from all pigs, which were then slaughtered to collect intestinal samples. We evidenced that LMWC not only increased (P < 0.05) the occludin protein abundance but also decreased (P < 0.05) the interleukin-6, tumour necrosis factor-α and mast cell tryptase contents, and the apoptotic epithelial cell percentages, in the small intestine of ETEC-infected pigs. Furthermore, LMWC down-regulated (P < 0.05) the small intestinal expression levels of critical inflammatory- and apoptotic-related genes, such as Toll-like receptor 4 (TLR4) and tumour necrosis factor receptor 1 (TNFR1), as well as the intra-nuclear nuclear factor-κB (NF-κB) p65 protein abundance, in the ETEC-infected pigs. Our study indicated a protective effect of LMWC on ETEC-triggered intestinal barrier disruption in weaned pigs, which involves the repression of intestinal inflammatory responses via blocking the TLR4/NF-κB signalling pathway and the depression of epithelial cell death via TNFR1-dependent apoptosis.


Assuntos
Anti-Inflamatórios/uso terapêutico , Apoptose , Quitosana/uso terapêutico , Infecções por Escherichia coli/tratamento farmacológico , Mucosa Intestinal/efeitos dos fármacos , Animais , Anti-Inflamatórios/farmacologia , Quitosana/farmacologia , Escherichia coli Enterotoxigênica , Interleucina-6/genética , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Ocludina/genética , Ocludina/metabolismo , Suínos , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/metabolismo , Fator de Transcrição RelA/genética , Fator de Transcrição RelA/metabolismo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo
20.
PLoS One ; 14(7): e0219211, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31276543

RESUMO

OBJECTIVE: Asphyxia of newborns is a severe and frequent challenge of the peri- and postnatal period. The purpose of this study was to study early morphological, immunological and structural alterations in lung tissue after asphyxia and hemorrhage (AH). METHODS: 44 neonatal piglets (age 32 hrs) underwent asphyxia and hemorrhage (AH) and were treated according to the international liaison committee of resuscitation (ILCOR) guidelines. For this study, 15 piglets (blood transfusion (RBC) n = 9; NaCl n = 6, mean age 31 hrs) were randomly picked. 4 hours after ROSC (return of spontaneous circulation), lung tissue and blood samples were collected. RESULTS: An elevation of myeloperoxidase (MPO) activity was observed 4 hrs after AH accompanied by an increase of surfactant D after RBC treatment. After AH tight junction proteins Claudin 18 and junctional adhesion molecule 1 (JAM1) were down-regulated, whereas Occludin was increased. Furthermore, after AH and RBC treatment dephosphorylated active form of Connexin 43 was increased. CONCLUSIONS: AH in neonatal pigs is associated with early lung injury, inflammation and alterations of tight junctions (Claudin, Occludin, JAM-1) and gap junctions (Connexin 43) in lung tissue, which contributes to the development of lung edema and impaired function.


Assuntos
Asfixia Neonatal/fisiopatologia , Lesão Pulmonar/fisiopatologia , Pulmão/fisiopatologia , Animais , Animais Recém-Nascidos/metabolismo , Asfixia/fisiopatologia , Asfixia Neonatal/metabolismo , Moléculas de Adesão Celular/metabolismo , Claudinas/metabolismo , Conexina 43/metabolismo , Modelos Animais de Doenças , Junções Comunicantes , Lesão Pulmonar/metabolismo , Ocludina/metabolismo , Peroxidase/análise , Proteína D Associada a Surfactante Pulmonar/análise , Choque Hemorrágico/imunologia , Choque Hemorrágico/metabolismo , Suínos , Junções Íntimas/metabolismo
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