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1.
J Agric Food Chem ; 67(35): 9789-9795, 2019 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-31373816

RESUMO

Pulmonary fibrosis is a chronic lung disease characterized by abnormal accumulation of the extracellular matrix (ECM). Chronic damage of the alveolar epithelium leads to a process called "epithelial-mesenchymal transition" (EMT) and increases synthesis and deposition of ECM proteins. Therefore, inhibition of EMT might be a promising therapeutic approach for the treatment of pulmonary fibrosis. ß-Sitosterol is one of the most abundant phytosterols in the plant kingdom and the major constituent in corn silk, which is derived from the stigma and style of maize (Zea mays). In this study, we elucidated that ß-sitosterol inhibited transforming growth factor-ß1 (TGF-ß1)-induced EMT and consequently had an antifibrotic effect. ß-Sitosterol (1-10 µg/mL) significantly downregulated the TGF-ß1-induced fibrotic proteins, such as collagen, fibronectin, and α-smooth muscle actin in human alveolar epithelial cells (p < 0.01). After 24 h, relative wound density (RWD) was increased in TGF-ß1 treated group (82.16 ± 5.70) compare to the control group (64.63 ± 2.21), but RWD was decreased in ß-sitosterol cotreated group (10 µg/mL: 71.54 ± 7.39; 20 µg/mL: 65.69 ± 6.42). In addition, the changes of the TGF-ß1-induced morphological shape and protein expression of EMT markers, N-cadherin, vimentin, and E-cadherin, were significantly blocked by ß-sitosterol treatment (p < 0.01). The effects of ß-sitosterol on EMT were found to be associated with the TGF-ß1/Snail pathway, which is regulated by Smad and non-Smad signaling pathways. Taken together, these findings suggest that ß-sitosterol can be used to attenuate pulmonary fibrosis through suppression of EMT by inhibiting the TGF-ß1/Snail pathway.


Assuntos
Células Epiteliais Alveolares/efeitos dos fármacos , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Extratos Vegetais/farmacologia , Alvéolos Pulmonares/efeitos dos fármacos , Fibrose Pulmonar/fisiopatologia , Sitosteroides/farmacologia , Zea mays/química , Actinas/genética , Actinas/metabolismo , Células Epiteliais Alveolares/citologia , Células Epiteliais Alveolares/metabolismo , Caderinas/genética , Caderinas/metabolismo , Linhagem Celular , Fibronectinas/genética , Fibronectinas/metabolismo , Humanos , Extratos Vegetais/química , Alvéolos Pulmonares/metabolismo , Alvéolos Pulmonares/fisiopatologia , Fibrose Pulmonar/tratamento farmacológico , Fibrose Pulmonar/genética , Fibrose Pulmonar/metabolismo , Transdução de Sinais/efeitos dos fármacos , Fator de Crescimento Transformador beta1/genética , Fator de Crescimento Transformador beta1/metabolismo
2.
Toxicol Lett ; 313: 178-187, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31284023

RESUMO

Long-term inhalation of crystalline silica particles leads to silicosis characterized by pulmonary inflammation and interstitial fibrosis. The growth arrest-specific protein 6 (Gas6) and its tyrosine receptor Mer have been implicated to involve in the regulation of inflammation, innate immunity and tissue repair. However, the role of Gas6 or Mer in silica-induced lung inflammation and fibrosis has not been investigated previously. In this study, we observed a remarkable increase of Gas6 in bronchoalveolar lavage fluid (BALF) from wild-type C57BL/6 mice after silica intratracheal administration. Then, we investigated whether genetic loss of Gas6 or Mer could attenuate silica-induced lung inflammation and fibrosis. Our results showed that Gas6-/- and Mer-/- mice exhibited reduced lung inflammation response from days 7 to 84 after silica exposure. We also uncovered an overexpression of the suppressor of cytokine signaling protein 1 in silica-treated deficient mice. Moreover, Gas6 or Mer deficiency attenuated silica-induced collagen deposition by inhibiting the expression of transforming growth factor-ß. We conclude that gene absence of Gas6 or Mer is protective against silica-induced lung inflammation and fibrosis in mice. Targeting Gas6/Mer pathway may be a potential therapeutic approach to treat pulmonary fibrosis in patients with silicosis.


Assuntos
Peptídeos e Proteínas de Sinalização Intercelular/deficiência , Pulmão/enzimologia , Pneumonia/prevenção & controle , Fibrose Pulmonar/prevenção & controle , Silicose/prevenção & controle , c-Mer Tirosina Quinase/deficiência , Animais , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Modelos Animais de Doenças , Peptídeos e Proteínas de Sinalização Intercelular/genética , Pulmão/patologia , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Pneumonia/enzimologia , Pneumonia/genética , Pneumonia/patologia , Fibrose Pulmonar/enzimologia , Fibrose Pulmonar/genética , Fibrose Pulmonar/patologia , Transdução de Sinais , Silicose/enzimologia , Silicose/genética , Silicose/patologia , Proteína 1 Supressora da Sinalização de Citocina/genética , Proteína 1 Supressora da Sinalização de Citocina/metabolismo , Fatores de Tempo , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/metabolismo , c-Mer Tirosina Quinase/genética
3.
Toxicol Lett ; 310: 7-13, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30978436

RESUMO

Silicosis is a kind of chronic and incurable lung fibrotic disease with pathogenesis and molecular mechanisms largely unknown. Mounting evidence suggests that long non-coding RNAs (lncRNAs) are involved in the pathogenesis of silicosis. However, how many lncRNAs involved in the pulmonary fibrosis remains to be elucidated. In this study, Wistar rats were exposed to silicon dioxide by an improved tracheal intubation method. Rats in the control group were treated with normal saline solution. Results showed that 28 days after exposure, there were significant differences in body weight and lung coefficient of rats treated with silica compared with control rats. The formation of lung fibrosis in silica-induced rats was confirmed by histologic examination. We then investigated the lncRNAs expression changes in lung tissues of silica-exposed rats and compared that with the rats in the control group using microarray. The results indicated that silica exposure leads to altered expression profile in 682 lncRNAs (300 upregulated and 382 downregulated). Seventy-three ceRNA pairs were acquired by predicted analysis. Kyoto Encyclopedia of Genes and Genomes pathway and Gene Ontology analyses were used to predict the biological pathway and functional classification of lncRNAs. The results showed that silica exposure affected 13 lncRNAs pathways. The functional classification mainly involved in protein binding, cell shape and extracellular exosome. This study indicated that alteration of lncRNAs may play a role in silica-induced pulmonary fibrosis through regulation of expressions of functional genes in lungs of rat. Our results provide more insights into the mechanism of silicosis.


Assuntos
Perfilação da Expressão Gênica/métodos , Pulmão/efeitos dos fármacos , Análise de Sequência com Séries de Oligonucleotídeos , Fibrose Pulmonar/genética , RNA Longo não Codificante/genética , Dióxido de Silício/toxicidade , Silicose/genética , Animais , Regulação da Expressão Gênica/efeitos dos fármacos , Redes Reguladoras de Genes/efeitos dos fármacos , Pulmão/metabolismo , Pulmão/patologia , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/metabolismo , Fibrose Pulmonar/patologia , RNA Longo não Codificante/metabolismo , Ratos Wistar , Silicose/metabolismo , Silicose/patologia , Fatores de Tempo , Transcriptoma/efeitos dos fármacos
4.
J Agric Food Chem ; 67(17): 4915-4922, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-31001980

RESUMO

Lung injury is a complicated and lethal condition characterized by alveolar barrier disruption, pulmonary edema, enhanced inflammation, and apoptosis in alveoli. However, therapeutic strategies to ameliorate lung injury without exerting side effects are not available. Functional amino acids have been shown to have anti-inflammatory and anti-apoptotic effects under various conditions. The objective of this study was to test the hypothesis that arginine, glutamine, or glycine supplementation ameliorated lipopolysaccharide (LPS)-induced lung injury in mice. Mice pretreated with aerosolized arginine, glutamine, or glycine were exposed to aerosolized LPS to induce lung injury. Results showed that arginine or glycine pretreatment beneficially reduced LPS-induced collagen deposition, apoptosis of alveolar cells, expression of inflammatory cytokines and chemokines, and accumulation of neutrophils and macrophages in lung tissues of mice, thus contributing to improved alveolar integrity and function. Glutamine administration reduced LPS-induced collagen deposition and inflammatory cytokines without affecting any other parameters examined in the study. Our findings indicated that arginine or glycine pretreatment effectively alleviated LPS-induced lung injury by inhibiting the accumulation of lymphocytes, the release of inflammatory cytokines and chemokines, and the apoptosis of alveolar cells. Supplementation of arginine or glycine may be a novel nutritional strategy to reduce deleterious effects of bacterial infection on alveolar function.


Assuntos
Aminoácidos/administração & dosagem , Apoptose/efeitos dos fármacos , Substâncias Protetoras/administração & dosagem , Fibrose Pulmonar/tratamento farmacológico , Animais , Quimiocinas/genética , Quimiocinas/imunologia , Citocinas/genética , Citocinas/imunologia , Humanos , Lipopolissacarídeos/efeitos adversos , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Pulmão/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neutrófilos/efeitos dos fármacos , Neutrófilos/imunologia , Fibrose Pulmonar/genética , Fibrose Pulmonar/imunologia , Fibrose Pulmonar/fisiopatologia
5.
Environ Toxicol ; 34(6): 728-741, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30815999

RESUMO

Pneumoconiosis is a serious occupational disease that often occurs to coal workers with no early diagnosis and effective treatment at present. Diffuse pulmonary fibrosis is the major pathological change of pneumoconiosis, and its mechanism is still unclear. Epigenetics is involved in the development of many diseases, and it is closely associated with fibrosis. In this study, we investigated whether DNA methylation contributes to the pathogenesis of pulmonary fibrosis in pneumoconiosis. By exposure to coal dust or silica dust, we established the models of coal worker's pneumoconiosis (CWP), which showed an increased expression of COL-I, COL-III. We further found that DNMT1, DNMT3a, DNMT3b, MBD2, MeCP2 protein expression changed. Pretreatment with DNMT inhibitor 5-aza-dC reduced expression of COL-I, COL-III, and reduced pulmonary fibrosis. In summary, our results showed that DNA methylation contributes to dust-induced pulmonary fibrosis and that it may serve as a theoretical basis for testing DNA methyltransferase inhibitors in the treatment of CWP.


Assuntos
Antracose/etiologia , Metilação de DNA/efeitos dos fármacos , Poeira , Epigênese Genética/efeitos dos fármacos , Fibrose Pulmonar/induzido quimicamente , Animais , Antracose/genética , Antracose/metabolismo , Linhagem Celular , Carvão Mineral/toxicidade , Minas de Carvão , Colágeno Tipo I/genética , Colágeno Tipo III/genética , DNA (Citosina-5-)-Metiltransferase 1/antagonistas & inibidores , DNA (Citosina-5-)-Metiltransferase 1/genética , Decitabina/farmacologia , Modelos Animais de Doenças , Humanos , Masculino , Exposição Ocupacional/efeitos adversos , Tamanho da Partícula , Fibrose Pulmonar/genética , Fibrose Pulmonar/metabolismo , Ratos Sprague-Dawley , Dióxido de Silício/química , Dióxido de Silício/toxicidade
6.
Eur J Pharmacol ; 851: 133-143, 2019 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-30797787

RESUMO

Rosiglitazone, a PPAR-γ agonist, possesses anti-fibritic effect; however, its inhibitory effect on paraquat (PQ)-induced pulmonary fibrosis is not completely understood. Here, we investigated the inhibitory effect of rosiglitazone on PQ-induced acute pulmonary fibrosis in rats and its underlying mechanism. Male Sprague-Dawly rats were administered a single intraperitoneal injection of 30 mg/kg PQ and euthanised 7, 14, 21, and 28 days after PQ poisoning. PQ-induced pulmonary fibrosis was most obvious on day 28. Male Sprague-Dawly rats were exposed either against distilled water as control groups or PQ (30 mg/kg, i.p.) as test groups. The control groups were nominated as NC group (without treatment), RSG group (only treatment with rosiglitazone, 10 mg/kg/d), and GW group (only treatment with GW9662, a PPAR-γ antagonist, 1 mg/kg/d). The test groups were nominated as PQ group (PQ exposed without treatment), PQ + RSG group (treatment with rosiglitazone), and PQ + RSG + GW group (treatment with rosiglitazone and GW9662). Rosiglitazone was able to recover the PQ-induced decrease in arterial oxygen partial pressure (PaO2), increase in the wet-to-dry (W/D) lung tissue weight ratio and lung fibrosis score. Rosiglitazone inhibited the PQ-induced reduction in protein and mRNA levels of PPAR-γ and PTEN and elevation in protein and mRNA levels of TGF-ß1 and α-SMA. GW9662 administration antagonized the effect of rosiglitazone. These data suggest that rosiglitazone attenuated PQ-induced pulmonary fibrosis by upregulateing PTEN and downregulating TGF-ß1 expression in a PPAR-γ dependent manner.


Assuntos
PPAR gama/metabolismo , Paraquat/envenenamento , Fibrose Pulmonar/tratamento farmacológico , Fibrose Pulmonar/metabolismo , Rosiglitazona/farmacologia , Actinas/genética , Animais , Aorta Abdominal/efeitos dos fármacos , Aorta Abdominal/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Hidroxiprolina/metabolismo , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Pulmão/patologia , Masculino , Tamanho do Órgão/efeitos dos fármacos , Oxigênio/metabolismo , PPAR gama/genética , PTEN Fosfo-Hidrolase/genética , PTEN Fosfo-Hidrolase/metabolismo , Fibrose Pulmonar/genética , Fibrose Pulmonar/patologia , Ratos , Ratos Sprague-Dawley , Rosiglitazona/uso terapêutico , Fator de Crescimento Transformador beta1/genética , Fator de Crescimento Transformador beta1/metabolismo
7.
Nat Genet ; 51(3): 494-505, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30804561

RESUMO

Chronic obstructive pulmonary disease (COPD) is the leading cause of respiratory mortality worldwide. Genetic risk loci provide new insights into disease pathogenesis. We performed a genome-wide association study in 35,735 cases and 222,076 controls from the UK Biobank and additional studies from the International COPD Genetics Consortium. We identified 82 loci associated with P < 5 × 10-8; 47 of these were previously described in association with either COPD or population-based measures of lung function. Of the remaining 35 new loci, 13 were associated with lung function in 79,055 individuals from the SpiroMeta consortium. Using gene expression and regulation data, we identified functional enrichment of COPD risk loci in lung tissue, smooth muscle, and several lung cell types. We found 14 COPD loci shared with either asthma or pulmonary fibrosis. COPD genetic risk loci clustered into groups based on associations with quantitative imaging features and comorbidities. Our analyses provide further support for the genetic susceptibility and heterogeneity of COPD.


Assuntos
Predisposição Genética para Doença/genética , Doença Pulmonar Obstrutiva Crônica/genética , Adulto , Idoso , Asma/genética , Estudos de Casos e Controles , Feminino , Expressão Gênica/genética , Loci Gênicos/genética , Estudo de Associação Genômica Ampla/métodos , Humanos , Pulmão/fisiopatologia , Masculino , Pessoa de Meia-Idade , Fenótipo , Polimorfismo de Nucleotídeo Único/genética , Fibrose Pulmonar/genética , Fumar/genética
8.
Biomed Res Int ; 2019: 6305065, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30756084

RESUMO

Objectives: As an epigenetic player, long noncoding RNAs (LncRNAs) have been reported to participate in multiple biological processes; however, their biological functions in silica-induced pulmonary fibrosis (SIPF) occurrence and development remain incompletely understood. Methods: Five case/control pairs were used to perform integrated transcriptomes analysis of lncRNA and mRNA. Prediction of lncRNA and mRNA functions was aided by the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. Additionally, we constructed a coexpression network of lncRNAs and mRNAs to identify targets of regulation. Results: In total, 1069 differentially expressed mRNAs and 366 lncRNAs were identified with the changes more than 2 times (p<0.05), of which 351 downregulated mRNA and 31 downregulated lncRNA were <0.5 (p<0.05) and those of 718 upregulated mRNAs and 335 upregulated lncRNA were >2 (p<0.05). The levels of 10 lncRNAs were measured via qRT-PCR; the results were consistent with the microarray data. Four genes named of FEM1B, TRIM39, TRIM32, and KLHL15 were enriched significantly with ubiquitination and immune response. Cytokine-cytokine receptor interaction was the most significantly enriched KEGG pathway in both mRNAs and lncRNAs. The coexpression network revealed that a single lncRNA can interact with multiple mRNAs, and vice versa. Conclusions: lncRNA and mRNA expression were aberrant in patients with SIPF compared to controls, indicating that differentially expressed lncRNAs and mRNAs may play critical roles in SIPF development. Our study affords new insights into the molecular mechanisms of SIPF and identifies potential biomarkers and targets for SIPF diagnosis and treatment.


Assuntos
Fibrose Pulmonar/genética , RNA Longo não Codificante/genética , RNA Mensageiro/genética , Ubiquitinação/genética , Idoso , Feminino , Perfilação da Expressão Gênica , Ontologia Genética , Humanos , Imunidade Inata/genética , Masculino , Pessoa de Meia-Idade , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/patologia , Transdução de Sinais/genética , Dióxido de Silício/toxicidade , Transcriptoma/genética
9.
Toxicology ; 414: 57-67, 2019 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-30658076

RESUMO

Various miRNAs are dysregulated during initiation and progression of pulmonary fibrosis. However, their function remains limited in silicosis. Here, we observed that miR-125a-3p was downregulated in silica-induced fibrotic murine lung tissues. Ectopic miR-125a-3p expression with chemotherapy attenuated silica-induced pulmonary fibrosis. Further in vitro experiments revealed that TGF-ß1 effectively decreased miR-125a-3p expression in fibroblast lines (NIH/3T3 and MRC-5). Overexpression of miR-125a-3p blocked fibroblast activation stimulated by TGF-ß1. Mechanistically, miR-125a-3p could bind to the 3'-untranslated region of Fyn and inhibit its expression in both mRNA and protein levels, thus causing inactivation of Fyn downstream effector STAT3. Fyn and p-STAT3, as opposed to miR-125a-3p expression, were elevated in silica-induced fibrotic murine lung tissues and TGF-ß1-treated fibroblast lines. Furthermore, Fyn knockdown or p-STAT3 suppression effectively attenuated fibroblast activation and ECM production. Taken together, miR-125a-3p is involved in fibrosis pathogenesis by fibroblast activation, suggesting that targeting miR-125a-3p/Fyn/STAT3 signaling pathway could be a potential therapeutic approach for pulmonary fibrosis.


Assuntos
Fibroblastos/enzimologia , Pulmão/enzimologia , MicroRNAs/metabolismo , Proteínas Proto-Oncogênicas c-fyn/metabolismo , Fibrose Pulmonar/enzimologia , Fator de Transcrição STAT3/metabolismo , Dióxido de Silício , Animais , Modelos Animais de Doenças , Fibroblastos/efeitos dos fármacos , Fibroblastos/patologia , Humanos , Pulmão/efeitos dos fármacos , Pulmão/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , MicroRNAs/genética , Células NIH 3T3 , Fenótipo , Fosforilação , Proteínas Proto-Oncogênicas c-fyn/genética , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/genética , Fibrose Pulmonar/patologia , Transdução de Sinais , Fator de Crescimento Transformador beta1/farmacologia
10.
Front Biosci (Landmark Ed) ; 24: 576-596, 2019 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-30468675

RESUMO

The molecular mechanism of how airway inflammation develops is not fully understood. Withaferin A (WA) is a natural phytochemical isolated from the plant Withania somnifera. It is a well-investigated bioactive compound that possesses a variety of health-promoting effects, including anti-inflammatory and anti-oxidative activities. In the present study, the effect of WA on ovalbumin (OVA)-induced airway inflammation in mice was investigated. The results indicated that pre-treatment with WA inhibited OVA-induced lung injury and fibrosis progression in mice. Furthermore, WA significantly downregulated inflammatory cell infiltration into the bronchoalveolar lavage fluid and significantly reduced pro-inflammatory cytokine expression in the lung tissue specimens. Additionally, WA significantly suppressed transforming growth factor-b1 expression in lung tissues. WA also caused the downregulation of collagen I, collagen III, α-smooth muscle actin and tissue inhibitor of metalloproteinase-1, as well as SMADs and extracellular signal related kinase 1/2 inactivation. Notably, WA significantly reduced the activation of the NLRP3 inflammasome. The results indicate that WA may be an effective novel candidate for the treatment of airway inflammation.


Assuntos
Inflamação/prevenção & controle , Ovalbumina/toxicidade , Sistema Respiratório/efeitos dos fármacos , Vitanolídeos/farmacologia , Animais , Linhagem Celular , Colágeno/genética , Colágeno/metabolismo , Citocinas/genética , Citocinas/metabolismo , Expressão Gênica/efeitos dos fármacos , Humanos , Inflamassomos/efeitos dos fármacos , Inflamassomos/genética , Inflamassomos/metabolismo , Inflamação/genética , Inflamação/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Fibrose Pulmonar/genética , Fibrose Pulmonar/metabolismo , Fibrose Pulmonar/prevenção & controle , Sistema Respiratório/metabolismo , Sistema Respiratório/patologia
11.
Life Sci ; 218: 241-252, 2019 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-30586565

RESUMO

Aberrantly activated Wnt signaling pathway and dysregulation of extracellular antagonists of Wnt signaling have been revealed in pulmonary fibrosis. In this study we evaluated the expression of secreted frizzled-related proteins (SFRPs) and their aberrant promoter methylation to investigate the involvement of epigenetic regulation in pulmonary fibrosis. The pulmonary fibrosis induced by intratracheal injection of bleomycin (BLM) into mice was adopted. The transcription and relative protein expression of SFRPs were detected at Day 7 (D7), D14, and D21. DNA methylation analysis was performed by methylation-specific polymerase chain reaction (MSP). A DNA methyltransferase (DNMT) inhibitor (5-aza-2'-deoxycytidine; 5-aza) was used for demethylation and the relative ß-catenin expression levels were measured to assess overactivity of the canonical Wnt signaling pathway. The transcription and protein expression of SFRP1 significantly decreased at D14 and D21, whereas the transcription and protein expression of SFRP4 significantly decreased at D7 and stayed downregulated until D21. The significantly hypermethylated promoters of SFRP1 and SFRP4 resulted in impaired transcription and decreased expression during pulmonary fibrosis in mice. Besides, reactivation of SFRP1 and SFRP4 by 5-aza reduced ß-catenin mRNA and protein expression in vivo and in vitro. Animal experiments confirmed that 5-aza could significantly alleviate bleomycin-induced pulmonary fibrosis in mice. Thus, changes of promoter hypermethylation might downregulate SFRP1 and SFRP4 at different stages of pulmonary fibrosis, and the finding supports the usefulness of DNMT inhibitors, which might effectively reverse activation of ß-catenin and reduce pulmonary fibrosis in mice. These data provide a possible new direction in the research on pulmonary fibrosis treatments.


Assuntos
Bleomicina/toxicidade , Metilação de DNA , Epigênese Genética , Peptídeos e Proteínas de Sinalização Intercelular/genética , Proteínas de Membrana/genética , Proteínas Proto-Oncogênicas/genética , Fibrose Pulmonar/genética , Fibrose Pulmonar/patologia , Animais , Antibióticos Antineoplásicos/toxicidade , Regulação para Baixo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Regiões Promotoras Genéticas , Fibrose Pulmonar/induzido quimicamente
12.
Chemosphere ; 219: 268-276, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30543962

RESUMO

SO2 and PAHs are well-known pollutants of coal burning and significant contributors to haze episodes. The purpose of the study is to determine whether the combined effects of SO2 and BaP are synergetic and to investigate the pro-fibrotic influences and possible mechanism from the aspect of microRNAs. In the present study cellular metabolic activity of BEAS-2B was assessed using MTT probe. C57BL/6 mice were exposed to BaP (40 mg/kg b.w.) for 5 days or SO2 (7 mg/m3) inhalation for 4 weeks alone or together. Lung tissues were processed for histology to assess pulmonary fibrosis. The protein level of pulmonary pro-fibrotic genes (Col1a1, Col3a1, alpha-SMA, fibronectin) and TGFßR2 were analyzed by Western blot and immunofluorescence in vivo and in vitro. Furthermore, we clarified that the microRNA expression of mir-30c-1-3p by real-time RT-PCR. The luciferase reporter assay was used to determine the binding sites of mir-30c-1-3p in the 3'-UTR of TGFßR2. It was confirmed that SO2 and BaP acted together to produce synergistic effects in cellular metabolic activity. Coexisting of SO2 and BaP increased the protein expression of pro-fibrotic genes and TGFßR2 and decreased mir-30c-1-3p in vivo and in vitro. Dual-luciferase reporter gene assays showed that TGFßR2 was a validated target of mir-30c-1-3p. All above results demonstrated that mir-30c-1-3p was involved in the synergistic pro-fibrotic effects of SO2 and BaP in lung via targeting TGFßR2. This work implies the potential risk of pulmonary fibrosis from the co-existence of SO2 and PAHs and provides new insights into the molecular markers for relevant diseases.


Assuntos
Benzopirenos/farmacologia , Camundongos Endogâmicos C57BL , MicroRNAs/metabolismo , Hidrocarbonetos Policíclicos Aromáticos/farmacologia , Fibrose Pulmonar/induzido quimicamente , Receptor do Fator de Crescimento Transformador beta Tipo II/metabolismo , Dióxido de Enxofre/farmacologia , Regiões 3' não Traduzidas , Animais , Sítios de Ligação , Camundongos , MicroRNAs/farmacologia , Fibrose Pulmonar/genética , Receptor do Fator de Crescimento Transformador beta Tipo II/análise , Receptor do Fator de Crescimento Transformador beta Tipo II/antagonistas & inibidores
14.
Medicine (Baltimore) ; 97(37): e11876, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30212926

RESUMO

Although many studies have investigated the association of single nucleotide polymorphisms (SNPs) in transforming growth factor beta1 (TGF-ß1) gene with pulmonary fibrosis (PF), but their association is still controversial. To clarify this, we performed a meta-analysis.Studies related to TGF-ß1 and PF were retrieved from PubMed, Medline, Embase, Scopus, and Wanfang (up to November 30, 2017). We targeted TGF-ß1 SNPs that have been reported by ≥3 studies to be included in the current meta-analysis, resulting in only 1 final SNP (rs1800470). The odds ratios (ORs) and 95% confidence intervals (CIs) were estimated in the models of allele comparison (T vs C), homozygote comparison (TT vs CC), dominant (TT vs TC + CC), recessive (TT + TC vs CC) to evaluate the strength of the associations.A total of 7 case-control studies were included in this meta-analysis. Overall, no significant association between TGF-ß1 rs1800470 and PF was found (T vs C: OR [95% CI] = 0.96 [0.80, 1.15]; TT vs CC: 0.87 [0.61, 1.22]; TT vs TC + CC: 0.80 [0.62, 1.04]; TT + TC vs CC: 1.13 [0.83, 1.54]). In subgroup analyses by ethnicity or original disease, no statistically significant association between TGF-ß1 rs1800470 polymorphisms and PF was demonstrated.This meta-analysis revealed that TGF-ß1 rs1800470 polymorphism was not associated with susceptibility to PF development.


Assuntos
Fibrose Pulmonar/genética , Fator de Crescimento Transformador beta1/genética , Alelos , Grupo com Ancestrais do Continente Asiático/genética , Estudos de Casos e Controles , Predisposição Genética para Doença , Genótipo , Humanos , Razão de Chances , Polimorfismo de Nucleotídeo Único , Fatores de Risco
15.
Gene ; 678: 252-260, 2018 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-30099020

RESUMO

OBJECTIVE: To investigate the regulatory mechanism of MEN1 gene in radiation-induced lung fibrosis in mice and provide a new theoretical basis for the clinical treatment of radiation pulmonary fibrosis. METHODS: First, 80 C57BL/6 mice aged 8 weeks and weighing 18-22 g were selected, half of them were male and the other half were female. The mice were divided into control group and irradiation group (40 mice in each group) according to the method of the random number table. A radiation-induced lung fibrosis mouse model was established in which a single X-ray irradiation of 20 Gy was applied to the right lung in the irradiation group; H&E and Masson staining were used to verify whether the model was successful at 4, 8, 16 and 24 weeks after irradiation. The expression of MEN1, smooth muscle actin (α-SMA), Collagen-1 and transforming growth factor (TGF-ß) in lung tissue were detected by Western blot and qPCR. Secondly, in the mouse embryonic fibroblast cell line (MEF) and mouse lung epithelial cell line (MLE-12), we constructed cell models of MEN1 knockout and interference separately with the irradiation of 10 Gy X-rays. The expression of α-SMA, Collagen-1, and TGF-ß/Smads signaling pathway molecules was detected by qPCR. Finally, using the immunoprecipitation (IP) method, we can detect the interaction between Smad2 and the protein menin encoded by the MEN1 gene. RESULTS: The results of the radiation pulmonary fibrosis model in mice showed that compared with the control group, the alveolar septum widens, the alveolar integrity decreases, the lung tissue slightly thickens, and a small amount of collagen deposits appear after 4-8 weeks in the model group. At twenty-fourth weeks, a large number of cells in the interstitial space of the lung tissue and a localized focal fibrosis area were observed. Further study found that radiation induced fibrogenic inflammatory cytokines TGF-ß up-regulation, down-regulation of MEN1 gene expression, and then enhanced the expression of α-SMA and promotes the transformation of fibroblasts to myofibroblasts; At the same time, the expression of Collagen-1 was enhanced, which suggested that the extracellular matrix was overconcentrated and eventually promoted the formation of pulmonary fibrosis. In vitro, we found that knockout and interference of MEN1 gene can significantly enhance radiation-induced fibrosis, and up-regulate the expression of downstream molecules Smad2 and Smad3 of TGF-ß signaling pathway, and down-regulate the expression of Smad7. Furthermore, it played an important role in regulating the process of radionuclide fibrosis. CONCLUSION: MEN1 plays a key role in the formation of pulmonary fibrosis by regulating the secretion of TGF-ß and the activation of TGF-ß/Smads signaling pathway.


Assuntos
Actinas/metabolismo , Colágeno Tipo I/metabolismo , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas/metabolismo , Fibrose Pulmonar/etiologia , Fator de Crescimento Transformador beta/metabolismo , Raios X/efeitos adversos , Actinas/genética , Animais , Linhagem Celular , Colágeno Tipo I/genética , Modelos Animais de Doenças , Feminino , Regulação da Expressão Gênica , Técnicas de Inativação de Genes , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fibrose Pulmonar/genética , Fibrose Pulmonar/metabolismo , Transdução de Sinais , Proteínas Smad/genética , Proteínas Smad/metabolismo , Fator de Crescimento Transformador beta/genética
16.
PLoS One ; 13(5): e0197937, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29813125

RESUMO

BACKGROUND: Integrin α8 (ITGA8) heterodimerizes with integrin ß1 and is highly expressed in stromal cells of the lung. Platelet-derived growth factor receptor beta (PDGFRß+) cells constitute a major population of contractile myofibroblasts in the lung following bleomycin-induced fibrosis. Integrin α8ß1 is upregulated in fibrotic foci in bleomycin-induced lung injury. However, the functional role of ITGA8 in fibrogenesis has not been characterized. In this study, we examined whether genetic deletion of ITGA8 from PDGFRß+ cells in the lung altered fibrosis. METHODS: Pdgfrb-Cre/+;Itga8flox/- or Pdgfrb-Cre/+;Itga8flox/flox (Cre+) and control mice (Cre-) were used for in vitro and in vivo studies. Primary cultures of PDGFRß+ cells were exposed to TGFß, followed by RNA isolation for qPCR. For in vivo studies, Cre+ and Cre- mice were characterized at baseline and after bleomycin-induced fibrosis. RESULTS: PDGFRß-selected cells from Cre+ animals showed higher levels of Col1a1 expression after treatment with TGFß. However, Cre- and Cre+ animals showed no significant difference in measures of acute lung injury or fibrosis following bleomycin challenge. CONCLUSION: While ITGA8 deletion in lung PDGFRß+ stromal cells showed evidence of greater Col1a1 mRNA expression after TGFß treatment in vitro, no functional difference was detected in vivo.


Assuntos
Cadeias alfa de Integrinas/metabolismo , Fibrose Pulmonar/metabolismo , Animais , Líquido da Lavagem Broncoalveolar/química , Modelos Animais de Doenças , Feminino , Deleção de Genes , Cadeias alfa de Integrinas/deficiência , Cadeias alfa de Integrinas/genética , Camundongos , Fibrose Pulmonar/genética , Regulação para Cima
17.
Medicine (Baltimore) ; 97(19): e0724, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29742735

RESUMO

RATIONALE: Dyskeratosis congenita (DC) is a rare inherited disease characterized by the classical mucocutaneous triad. Pulmonary fibrosis, bone marrow failure, and solid tumors are the main causes of mortality in DC. Pathogenic variants in TERT, TERC, and DKC1 have been identified in individuals with familial pulmonary fibrosis. Mutations in TINF2 gene have been reported to be associated with bone marrow failure in most cases. However, the relationship between TINF2 mutation and pulmonary fibrosis is not yet clear. PATIENT CONCERNS: Here, we report the case of a 32-year-old woman presented with irritating cough for 2 years and progressive breathlessness for 6 months. DIAGNOSES: The patient was diagnosed with DC based on the following clinical evidences. Along with some family members, she had the typical mucocutaneous triad and pulmonary fibrosis. A heterozygous mutation (c.844C>T), located in exon 6 of TINF2 gene, that changed arginine to cysteine (Arg282Cys) was identified in this proband by whole exome sequencing. INTERVENTIONS: The patient received corticosteroid therapy but refused to receive lung transplantation. OUTCOMES: The proband died of respiratory failure 4 months after the diagnosis. The missense mutation was located in the conserved region of TINF2 gene and predicted to be deleterious by altering the protein structure. LESSONS: Lung transplantation should be considered for improved survival of patients with DC, and pulmonary fibrosis. Whole exome and whole genome sequencing should be widely used in the identification of such rare genetic variants for clinical diagnosis. The study of DC with pulmonary fibrosis can provide a more appropriate means of clinical research and therapy to the unfortunate patients who suffer from this rare disorder.


Assuntos
Disceratose Congênita/genética , Mutação de Sentido Incorreto , Fibrose Pulmonar/genética , Proteínas de Ligação a Telômeros/genética , Corticosteroides/uso terapêutico , Adulto , Disceratose Congênita/complicações , Evolução Fatal , Feminino , Humanos , Linhagem , Fibrose Pulmonar/complicações , Fibrose Pulmonar/tratamento farmacológico
18.
Mol Med Rep ; 17(6): 7781-7788, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29620190

RESUMO

Pulmonary fibrosis is a severe respiratory disease characterized by the aggregation of extracellular matrix components and inflammation­associated injury. Studies have suggested that long non­coding RNAs (lncRNA) may serve a role in the pathophysiological processes of pulmonary fibrosis. However, the potential molecular mechanisms involving the lncRNA, prostate cancer­associated transcript 29 (lncRNAPCAT29) in the progression of pulmonary fibrosis are yet to be determined. In the present study, the role of lncRNAPCAT29 and the potential signaling mechanism in pulmonary fibrosis progression was investigated. Reverse transcription­quantitative polymerase chain reaction and immunohistochemistry revealed that the expression levels of lncRNAPCAT29 were downregulated within interstitial lung cells from mice with silica­induced pulmonary fibrosis. Transfection with lncRNAPCAT29 was associated with upregulated expression of microRNA (miRNA)­221 and downregulated expression of transforming growth factor­ß1 (TGF­ß1); reduced inflammation and fibrotic progression was also associated with lncRNAPCAT29 transfection. TGF­ß1 expression levels were inhibited within pulmonary fibroblasts due to lncRNAPCAT29 expression; NEDD4 binding protein 2 and Plexin­A4 expression levels were also suppressed. Analysis of the potential mechanism underlying silica­induced pulmonary fibrosis revealed that the expression levels of RAS protein activator like 1 (RASAL1) and extracellular signal­regulated kinases 1/2 (ERK1/2) were suppressed due to lncRNAPCAT29 expression. The results of the present study demonstrated that lncRNAPCAT29 induced miRNA­221 upregulation and TGF­ß1 downregulation. These observations were associated with reduced inflammation and progression of silica­induced pulmonary fibrosis via the TGF­ß1­regulated RASAL1/ERK1/2 signaling pathway, which may serve as a potential target for the treatment of pulmonary fibrosis.


Assuntos
Proteínas Ativadoras de GTPase/metabolismo , Sistema de Sinalização das MAP Quinases , Fibrose Pulmonar/genética , Fibrose Pulmonar/metabolismo , RNA Longo não Codificante/genética , Fator de Crescimento Transformador beta1/metabolismo , Animais , Diferenciação Celular , Movimento Celular , Proliferação de Células , Citocinas/metabolismo , Enzimas Reparadoras do DNA/metabolismo , Fibroblastos/citologia , Fibroblastos/metabolismo , Humanos , Mediadores da Inflamação/metabolismo , Masculino , Camundongos , Fibrose Pulmonar/patologia , Transdução de Sinais , Fator de Crescimento Transformador beta1/genética
19.
Int J Mol Sci ; 19(4)2018 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-29677166

RESUMO

The administration of Everolimus (EVE), a mTOR inhibitor used in transplantation and cancer, is often associated with adverse effects including pulmonary fibrosis. Although the underlying mechanism is not fully clarified, this condition could be in part caused by epithelial to mesenchymal transition (EMT) of airway cells. To improve our knowledge, primary bronchial epithelial cells (BE63/3) were treated with EVE (5 and 100 nM) for 24 h. EMT markers (α-SMA, vimentin, fibronectin) were measured by RT-PCR. Transepithelial resistance was measured by Millicell-ERS ohmmeter. mRNA and microRNA profiling were performed by Illumina and Agilent kit, respectively. Only high dose EVE increased EMT markers and reduced the transepithelial resistance of BE63/3. Bioinformatics showed 125 de-regulated genes that, according to enrichment analysis, were implicated in collagen synthesis/metabolism. Connective tissue growth factor (CTGF) was one of the higher up-regulated mRNA. Five nM EVE was ineffective on the pro-fibrotic machinery. Additionally, 3 miRNAs resulted hyper-expressed after 100 nM EVE and able to regulate 31 of the genes selected by the transcriptomic analysis (including CTGF). RT-PCR and western blot for MMP12 and CTGF validated high-throughput results. Our results revealed a complex biological network implicated in EVE-related pulmonary fibrosis and underlined new potential disease biomarkers and therapeutic targets.


Assuntos
Antineoplásicos/efeitos adversos , Everolimo/efeitos adversos , MicroRNAs/genética , Fibrose Pulmonar/metabolismo , Transcriptoma/genética , Actinas/metabolismo , Animais , Linhagem Celular , Células Cultivadas , Biologia Computacional , Fator de Crescimento do Tecido Conjuntivo/metabolismo , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Fibronectinas/metabolismo , Humanos , Metaloproteinase 12 da Matriz/metabolismo , Camundongos , Células NIH 3T3 , Fibrose Pulmonar/genética , RNA Mensageiro/metabolismo
20.
Biochem Biophys Res Commun ; 500(4): 839-845, 2018 06 12.
Artigo em Inglês | MEDLINE | ID: mdl-29704504

RESUMO

Epithelial-mesenchymal transition (EMT) plays a pivotal role in idiopathic pulmonary fibrosis (IPF). In bleomycin-induced pulmonary fibrosis mice, we observed that inhibition of mTOR (mammalia target of rapamycin) attenuated IPF. Rapamycin suppressed the down-regulation of E-cadherin and up-regulation of fibronectin in bleomycin-induced pulmonary fibrosis mice. In addition, dual immunofluorescence staining for E-cadherin and fibronectin demonstrated that rapamycin pretreatment decreased the proportions of AECs undergoing EMT in bleomycin-induced pulmonary fibrosis, indicating that mTOR inhibition suppressed EMT in vivo. In the setting of transforming growth factor (TGF)-ß1-induced EMT in AECs, we found that mTOR inhibitor attenuated TGF-ß1-induced EMT in AECs. This EMT was characterized by morphology and cell skeleton changes and the expression of EMT phenotype markers. Finally, mTOR blockade decreased S6k and TGF-ß1-induced Smad2/3 phosphorylation. Bleomycin induced pulmonary fibrosis and EMT in mice, while mTOR repression inhibited bleomycin-induced pulmonary fibrosis and attenuated EMT in vivo. Hence, our study provided evidence of a novel mechanism by which mTOR inhibitor ameliorates pulmonary fibrosis. Suppression of mTOR and EMT may be a target for treatment of pulmonary fibrosis.


Assuntos
Transição Epitelial-Mesenquimal/efeitos dos fármacos , Alvéolos Pulmonares/efeitos dos fármacos , Fibrose Pulmonar/tratamento farmacológico , Sirolimo/farmacologia , Serina-Treonina Quinases TOR/genética , Animais , Bleomicina , Caderinas/genética , Caderinas/metabolismo , Modelos Animais de Doenças , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Transição Epitelial-Mesenquimal/genética , Fibronectinas/genética , Fibronectinas/metabolismo , Regulação da Expressão Gênica , Masculino , Camundongos , Fosforilação/efeitos dos fármacos , Alvéolos Pulmonares/metabolismo , Alvéolos Pulmonares/patologia , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/genética , Fibrose Pulmonar/patologia , Proteínas Quinases S6 Ribossômicas 90-kDa/antagonistas & inibidores , Proteínas Quinases S6 Ribossômicas 90-kDa/genética , Proteínas Quinases S6 Ribossômicas 90-kDa/metabolismo , Transdução de Sinais , Proteína Smad2/antagonistas & inibidores , Proteína Smad2/genética , Proteína Smad2/metabolismo , Proteína Smad3/antagonistas & inibidores , Proteína Smad3/genética , Proteína Smad3/metabolismo , Serina-Treonina Quinases TOR/antagonistas & inibidores , Serina-Treonina Quinases TOR/metabolismo , Fator de Crescimento Transformador beta1/antagonistas & inibidores , Fator de Crescimento Transformador beta1/genética , Fator de Crescimento Transformador beta1/metabolismo
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