RESUMO
Silicosis caused by inhalation of silica particles leads to more than ten thousand new occupational exposure-related deaths yearly. Exacerbating this issue, there are currently few drugs reported to effectively treat silicosis. Tetrandrine is the only drug approved for silicosis treatment in China, and despite more than decades of use, its efficacy and mechanisms of action remain largely unknown. Here, in this study, we established silicosis mouse models to investigate the effectiveness of tetrandrine of early and late therapeutic administration. To this end, we used multiple cardiopulmonary function test, as well as markers for inflammation and fibrosis. Moreover, using single cell RNA sequencing and transcriptomics of lung tissue and quantitative microarray analysis of serum from silicosis and control mice, our results provide a novel description of the target pathways for tetrandrine. Specifically, we found that tetrandrine attenuated silicosis by inhibiting both the canonical and non-canonical NLRP3 inflammasome pathways in lung macrophages. Taken together, our work showed that tetrandrine yielded promising results against silicosis-associated inflammation and fibrosis and further lied the groundwork for understanding its molecular targets. Our results also facilitated the wider adoption and development of tetrandirne, potentially accelerating a globally accepted therapeutic strategy for silicosis.
Assuntos
Inflamassomos , Silicose , Animais , Benzilisoquinolinas , Fibrose , Inflamassomos/metabolismo , Inflamação/metabolismo , Pulmão/patologia , Macrófagos/metabolismo , Camundongos , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Silicose/tratamento farmacológico , Silicose/metabolismoRESUMO
Silicosis is a global occupational disease characterized by lung dysfunction, pulmonary inflammation, and fibrosis, for which there is a lack of effective drugs. Pirfenidone has been shown to exert anti-inflammatory and anti-fibrotic properties in the lung. However, whether and how pirfenidone is effective against silicosis remains unknown. Here, we evaluated the efficacy of pirfenidone in the treatment of early and advanced silicosis in an experimental mouse model and explored its potential pharmacological mechanisms. We found that pirfenidone alleviated silica-induced lung dysfunction, secretion of inflammatory cytokines (TNF-α, IL-1ß, IL-6) and deposition of fibrotic proteins (collagen I and fibronectin) in both early and advanced silicosis models. Moreover, we observed that both 100 and 200 mg/kg pirfenidone can effectively treat early-stage silicosis, while 400 mg/kg was recommended for advanced silicosis. Mechanistically, antibody array and bioinformatic analysis showed that the pathways related to IL-17 secretion, including JAK-STAT pathway, Th17 differentiation, and IL-17 pathway, might be involved in the treatment of silicosis by pirfenidone. Further in vivo experiments confirmed that pirfenidone reduced the production of IL-17A induced by silica exposure via inhibiting STAT3 phosphorylation. Neutralizing IL-17A by anti-IL-17A antibody improved lung function and reduced pulmonary inflammation and fibrosis in silicosis animals. Collectively, our study has demonstrated that pirfenidone effectively ameliorated silica-induced lung dysfunction, pulmonary inflammation and fibrosis in mouse models by inhibiting the secretion of IL-17A.
Assuntos
Interleucina-17 , Pneumonia , Animais , Modelos Animais de Doenças , Fibrose , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Interleucina-17/metabolismo , Janus Quinases/metabolismo , Janus Quinases/uso terapêutico , Pulmão/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Pneumonia/induzido quimicamente , Pneumonia/tratamento farmacológico , Pneumonia/metabolismo , Piridonas , Fatores de Transcrição STAT/metabolismo , Fatores de Transcrição STAT/uso terapêutico , Transdução de Sinais , Dióxido de Silício/toxicidadeRESUMO
Object: By retrospectively analyzing the energy spectrum of squamous cell carcinoma, adenocarcinoma, small cell lung cancer (SCLC), and pulmonary metastases that underwent dual-layer detector spectral computed tomography (DLCT) 3-phase scan of the chest, we explored the value of a multiparameter energy spectrum in the assessment of pathological types of lung tumors. Methods: Cases of squamous cell carcinoma (n = 20), adenocarcinoma (n = 24), SCLC (n = 26), and metastases (n = 14) were collected. Then the largest cross-sectional area (LCA) of the lesion, computed tomography (CT) values in the plain scan phase, arterial and venous phases (HU, HUa, and HUv), iodine concentration, and effective atomic number in the arterial and venous phases (ICa, ICv, Zeff[a], and Zeff[v]) were measured and compared among the nonsmall cell lung cancer (NSCLC), SCLC and metastases, and other 3 groups of SCLC, squamous cell carcinoma, and adenocarcinoma. Results: Only the LCA is statistically different among SCLC, NSCLC, and metastases (P < .05). And the treated subgroup analysis did not show significant differences among the groups. However, the untreated subgroup analysis showed that there was a significant difference between NSCLC and metastases in LCA, SCLC and metastases in ICa, NSCLC and SCLC in HUv, NSCLC and SCLC in Zeff(v) (P < .05). Conclusion: The energy spectrum parameters of DLCT have a certain clinical value in distinguishing NSCLC from SCLC in the Zeff(v) and distinguishing SCLC from metastases in the ICa.
Assuntos
Neoplasias Pulmonares/diagnóstico por imagem , Neoplasias Pulmonares/patologia , Tomografia Computadorizada por Raios X/métodos , Idoso , Tomada de Decisão Clínica , Diagnóstico Diferencial , Gerenciamento Clínico , Feminino , Humanos , Processamento de Imagem Assistida por Computador , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Estudos Retrospectivos , Tomografia Computadorizada por Raios X/normasRESUMO
Pneumocystis jirovecii is an opportunistic fungus that can cause severe and potentially fatal Pneumocystis pneumonia (PCP) in immunodeficient patients. In this study, we investigated the genetic polymorphisms of P. jirovecii at eight different loci, including six nuclear genes (ITS, 26S rRNA, sod, dhps, dhfr and ß-Tub) and two mitochondrial genes (mtLSU-rRNA and cyb) in three PCP cases, including two patients with HIV infection and one without HIV infection in Shanxi Province, P.R. China. The gene targets were amplified by PCR followed by sequencing of plasmid clones. The HIV-negative patient showed a coinfection with two genotypes of P. jirovecii at six of the eight loci sequenced. Of the two HIV-positive patients, one showed a coinfection with two genotypes of P. jirovecii at the same two of the six loci as in the HIV-negative patient, while the other showed a single infection at all eight loci sequenced. None of the three drug target genes (dhfr, dhps and cyb) showed mutations known to be potentially associated with drug resistance. This is the first report of genetic polymorphisms of P. jirovecii in PCP patients in Shanxi Province, China. Our findings expand our understanding of the genetic diversity of P. jirovecii in China.
Assuntos
Infecções por HIV , Pneumocystis carinii , Pneumonia por Pneumocystis , Infecções Oportunistas Relacionadas com a AIDS/microbiologia , China , Coinfecção , Infecções por HIV/complicações , Infecções por HIV/microbiologia , Humanos , Pneumocystis carinii/genética , Pneumonia por Pneumocystis/microbiologia , Polimorfismo GenéticoRESUMO
Club cell protein (CC16) is expressed primarily by club cells possesses antiinflammatory properties and is located in the bronchiolar epithelium. Previous studies have demonstrated that CC16 deficiency is associated with the progression of chronic obstructive pulmonary disease (COPD). In the present study, the therapeutic effects of recombinant rat CC16 protein in mice with COPD were examined and the underlying mechanisms investigated. A total of 30 adult male C57/BL6 mice were randomly divided into three groups (10 mice/group). A mouse COPD model was generated by exposing 20 mice to cigarette smoke (CS) for 24 weeks. A total of 10 mice were treated intranasally with rCC16 (2.5 µg/g body weight) and control mice were exposed to normal room air. Results indicated that rCC16 treatment ameliorated pathological damage in the lungs and reduced the production of tumor necrosis factor (TNF)α, interleukin (IL)6 and IL8, which were induced by CS exposure. After rCC16 administration, endogenous CC16 was upregulated and the body weight of COPD mice was increased, whereas the opposite was observed in CSexposed mice. Additionally, rCC16 treatment inhibited the DNA binding of NFκB/p65 in lung tissues and reduced nuclear translocation of NFκB/p65 in BALF and epithelial cells. Moreover, rCC16 treatment lead to a decrease in the total number of BALF cells, including macrophages, which was elevated in COPD mice. In conclusion, the present results demonstrate that rCC16 has therapeutic effects on COPD by downregulating proinflammatory factors via the NFκB pathway.
Assuntos
Fumar Cigarros/metabolismo , Pneumonia/metabolismo , Doença Pulmonar Obstrutiva Crônica/metabolismo , Mucosa Respiratória/metabolismo , Transdução de Sinais/efeitos dos fármacos , Uteroglobina/farmacologia , Animais , Fumar Cigarros/tratamento farmacológico , Fumar Cigarros/patologia , Citocinas/metabolismo , Modelos Animais de Doenças , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Inflamação/patologia , Masculino , Camundongos , Pneumonia/tratamento farmacológico , Pneumonia/patologia , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Doença Pulmonar Obstrutiva Crônica/patologia , Proteínas Recombinantes/farmacologia , Mucosa Respiratória/patologia , Fator de Transcrição RelA/metabolismoRESUMO
Clara cell protein (CC16) is an anti-inflammatory protein, which is expressed in the airway epithelium. It is involved in the development of airway inflammatory diseases, including chronic obstructive pulmonary disease and asthma. However, the exact molecular mechanism underlying its antiinflammatory action remains to be fully elucidated. The aim of the present study was to define the protein profiles of the antiinflammatory effect of CC16 in lipopolysaccharide (LPS)treated rat tracheal epithelial (RTE) cells using shotgun proteomics. Protein extracts were obtained from control RTE cells, RTE cells treated with LPS and RTE cells treated with LPS and recombinant CC16 (rCC16). Subsequent labelfree quantification and bioinformatics analyses identified 12 proteins that were differentially expressed in the three treatment groups as a cluster of five distinct groups according to their molecular functions. Five of the twelve proteins were revealed to be associated with the cytoskeleton: Matrix metalloproteinase9, myosin heavy chain 10, actinrelated protein3 homolog, elongation factor 1α1 (EF1α1), and acidic ribosomal phosphoprotein P0. Five of the twelve proteins were associated with cellular proliferation: DNAdependent protein kinase catalytic subunit, EF1α1, tyrosine 3monooxygenase, caspase recruitment domain (CARD) protein 12 and adenosylhomocysteinase (SAHH) 3. Three proteins were associated with gene regulation: EF1α1, SAHH 3 and acidic ribosomal phosphoprotein P0. Three proteins were associated with inflammation: Tyrosine 3monooxygenase, CARD protein 12 and statinrelated protein. ATPase (H+transporting, V1 subunit A, isoform 1) was revealed to be associated with energy metabolism, and uridine diphosphate glycosyltransferase 1 family polypeptide A8 with drug metabolism and detoxification. The identified proteins were further validated using reverse transcriptionquantitative polymerase chain reaction. These protein profiles, and their interacting protein network, may facilitate the elucidation of the molecular mechanisms underlying the antiinflammatory effects of CC16.
Assuntos
Células Epiteliais/metabolismo , Inflamação/genética , Proteínas Recombinantes/genética , Traqueia/metabolismo , Uteroglobina/genética , Animais , Cromatografia Líquida , Células Epiteliais/patologia , Humanos , Inflamação/induzido quimicamente , Inflamação/metabolismo , Inflamação/patologia , Lipopolissacarídeos/toxicidade , Biossíntese de Proteínas/genética , Proteômica , Doença Pulmonar Obstrutiva Crônica , Ratos , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/metabolismo , Espectrometria de Massas em Tandem , Traqueia/patologia , Uteroglobina/administração & dosagem , Uteroglobina/metabolismoRESUMO
OBJECTIVE: To probe further the role of neutrophil activation in the lung injury induced by mechanical ventilation. METHODS: Thirty-two normal Wistar rats were randomly divided into four groups: control group, low tidal volume group, conventional tidal volume group, and high tidal volume group. The counts of white blood cell (WBC) and neutrophil in bronchoalveolar lavage fluid (BALF) were carried out. The levels of protein and myeloperoxidase (MPO) activity in plasma and BALF were determined by biochemical methods respectively. RESULTS: The numbers of WBC and neutrophils, and the levels of protein and MPO activity in BALF were significantly increased in normal and high tidal volume groups than those in control and low tidal volume groups (P<0.05 or P<0.01, respectively). The levels of protein and MPO activity in BALF in high tidal volume group were markedly higher compared with normal tidal volume group (both P<0.01), but no statistical differences existed between the low tidal volume group and the control group (both P>0.05). There were no significant differences in the levels of protein and MPO activity in plasma among the four groups (P>0.05). CONCLUSION: The recruitment and activation of neutrophils might play an important role in pathogenesis of the biotrauma induced by mechanical ventilation. MPO activity in BALF is a credible index in reflecting the activation of neutrophils. Measuring the level of protein in BALF is valuable to evaluate the extent of lung injury induced by mechanical ventilation.
Assuntos
Ativação de Neutrófilo , Lesão Pulmonar Induzida por Ventilação Mecânica/imunologia , Animais , Líquido da Lavagem Broncoalveolar , Modelos Animais de Doenças , Masculino , Peroxidase/metabolismo , Distribuição Aleatória , Ratos , Ratos Wistar , Lesão Pulmonar Induzida por Ventilação Mecânica/enzimologiaRESUMO
MicroRNAs are small noncoding RNA molecules that regulate gene expression at the post-transcriptional level. Compelling evidence reveals that there is a causative link between microRNAs deregulation and lung cancer development and metastasis. The aim of present study was to explore the function of miR-140-3p in the development and metastasis of lung cancer cell. Using real-time PCR, we detected the miR-140-3p expression of lung cancer tissues and its pared non-lung cancer tissue. Then, we evaluated the role of miR-140-3p in cell proliferation, invasion and migration using MTT, colony formation assay, Transwell invasion and Transwell migration assay in lung cancer cell lines. As a result, miR-140-3p expression level was lower in lung cancer tissues compared to adjacent normal lung cancer tissue. After miR-140-3p was upregulated in A549 or H1299 cells, cell proliferation, invasion and migration was notably attenuated. Furthermore, we identified ATP6AP2, which is associated with adenosine triphosphatases (ATPases), was a directly target of miR-140-3p in lung cancer cells. In conclusion, our data suggest miR-140-3p/ATP6AP2 axis might act as a potential therapeutic biomarker for lung cancer.
Assuntos
Movimento Celular/genética , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica , Neoplasias Pulmonares/metabolismo , MicroRNAs/metabolismo , Invasividade Neoplásica/patologia , Receptores de Superfície Celular/metabolismo , ATPases Vacuolares Próton-Translocadoras/metabolismo , Linhagem Celular Tumoral , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , MicroRNAs/genética , Invasividade Neoplásica/genética , Receptores de Superfície Celular/genética , Regulação para Cima , ATPases Vacuolares Próton-Translocadoras/genéticaRESUMO
BACKGROUND: Ventilator induced lung injury (VILI) is a serious complication in the treatment of mechanical ventilating patients, and it is also the main cause that results in exacerbation or death of patients. In this study, we produced VILI models by using glucocorticoid in rats with high tidal volume mechanical ventilation, and observed the content of macrophage inflammatory protein-1α (MIP-1α) in plasma and bronchoalveolar lavage fluid (BALF) and the expression of MIP-1α mRNA and nuclear factor-kappa B (NF-κB) p65 mRNA in the lung so as to explore the role of glucocorticoid in mechanical ventilation. METHODS: Thirty-two healthy Wistar rats were randomly divided into a control group, a ventilator induced lung injury (VILI) group, a dexamethasone (DEX) group and a budesonide (BUD) group. The content of MIP-1α in plasma and BALF was measured with ELISA and the level of MIP-1α mRNA and NF-κBp65 mRNA expressing in the lung of rats were detected by RT-PCR. The data were expressed as mean±SD and were compared between the groups. RESULTS: The content of MIP-1α in plasma and BALF and the level of MIP-1α mRNA and NF-KBp65 mRNA in the lung in the DEX and BUD groups were significantly lower than those in the VILI group (P<0.001). Although the content of MIP-1α in plasma and BALF and the level of MIP-1α mRNA and NF-κBp65 mRNA in the lung in the BUD group were higher than those in the DEX group, there were no significant differences between them (P>0.05). CONCLUSIONS: Glucocorticoid could down-regulate the expression of MIP-1α by inhibiting the activity of NF-κB in the lung and may exert preventive and therapeutic effects on VILI to some extent. The effect of local use of glucocorticoid against VILI is similar to that of systemic use, but there is lesser adverse reaction.