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1.
Toxicol Lett ; 319: 119-128, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31682869

RESUMO

Long-term exposure to fine particulate matter (PM2.5) may cause or exacerbate many diseases, including respiratory inflammation. However, the full mechanism is not yet fully understood. The newly discovered long chain non-coding RNA, though unable to encode proteins, regulates multiple life activities and participates in the development of inflammation. In this study, we set up a cell inflammation model by using normal bronchial 16HBE cells exposed to PM2.5. High-throughput sequencing, as well as real-time fluorescent quantitative PCR detection and validation, was performed on the inflamed cells to evaluate the expression level of long chain noncoding RNA that helped us to identify the LncRNA LOC101927514. Inhibiting LncRNA LOC101927514 expression by RNAi, reflected in a reduction in inflammation, is driven by PM2.5. In addition, we identify LncRNA LOC101927514 to be located within the nucleus and binds to STAT3, altering the inflammatory state of the cells and IL6 and IL8 release. This study identifies that LncRNA LOC101927514 is a new potential target for future treatment of the inflammatory response activated by PM2.5 in the respiratory system.


Assuntos
Poluentes Atmosféricos/toxicidade , Brônquios/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Inflamação/induzido quimicamente , Inflamação/genética , Material Particulado/toxicidade , RNA Longo não Codificante/genética , Fator de Transcrição STAT3/metabolismo , Contagem de Células , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Humanos , Interleucina-6/biossíntese , Interleucina-6/genética , Interleucina-8/biossíntese , Interleucina-8/genética , Fosforilação , Ligação Proteica , RNA Longo não Codificante/metabolismo
2.
Toxicol Lett ; 319: 256-263, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31639410

RESUMO

Transcription factor activator protein (AP)-1 can be activated in nitrogen-mustard-injured mouse skin, and is thought to participate in the inflammatory response. AP-1 consists of homo- or heterodimers of Fos [c-Fos, Fos-B, fos-related antigen (Fra)-1 and Fra-2] and Jun (c-Jun, JunB and JunD) family members, and information about their expression, location and function are still unclear. In nitrogen-mustard-exposed mouse skin, we found p-ERK activation increased Fra-1 and FosB. Unlike the nucleus location of c-Fos and FosB, Fra-1 and Fra-2 were mainly expressed in the cytoplasm. In nitrogen-mustard-exposed cultured immortalized human keratinocytes (HaCaT cells), Fra-1 in the nucleus functioned as an inhibitor of inflammatory cytokine interleukin (IL)-8. Co-immunoprecipitation showed that Fra-1 formed dimers with IL-8 transcription factors c-Jun, JunB and JunD. Fra-1 depletion increased c-Fos and FosB in the nucleus, accompanied by increased heterodimers of c-Fos/c-Jun, c-Fos/JunB, c-Fos/JunD, and FosB/JunB. In conclusion, Fra-1 trapped in the cytoplasm after nitrogen mustard exposure might be a driving force for IL-8 over-expression in injured skin.


Assuntos
Substâncias para a Guerra Química/toxicidade , Epiderme/lesões , Epiderme/metabolismo , Interleucina-8/biossíntese , Mecloretamina/toxicidade , Proteínas Proto-Oncogênicas c-fos/metabolismo , Animais , Linhagem Celular , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Citoplasma/efeitos dos fármacos , Citoplasma/metabolismo , Humanos , Queratinócitos/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Masculino , Camundongos , Camundongos Pelados , RNA Interferente Pequeno/farmacologia
3.
J Med Microbiol ; 68(12): 1813-1822, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31674896

RESUMO

Introduction. Chronic pulmonary infection is associated with colonization with multiple micro-organisms but host-microbe and microbe-microbe interactions are poorly understood.Aim. This study aims to investigate the differences in host responses to mono- and co-infection with S. aureus and B. cenocepacia in human airway epithelial cells.Methodology. We assessed the effect of co-infection with B. cenocepacia and S. aureus on host signalling and inflammatory responses in the human airway epithelial cell line 16HBE, using ELISA and western blot analysis.Results. The results show that B. cenocepacia activates MAPK and NF-κB signalling pathways, subsequently eliciting robust interleukin (IL)-8 production. However, when airway epithelial cells were co-treated with live B. cenocepacia bacteria and S. aureus supernatants (conditioned medium), the pro-inflammatory response was attenuated. This anti-inflammatory effect was widely exhibited in the S. aureus isolates tested and was mediated via reduced MAPK and NF-κB signalling, but not via IL-1 receptor or tumour necrosis factor receptor modulation. The staphylococcal effectors were characterized as small, heat-stable, non-proteinaceous and not cell wall-related factors.Conclusion. This study demonstrates for the first time the host response in a S. aureus/B. cenocepacia co-infection model and provides insight into a staphylococcal immune evasion mechanism, as well as a therapeutic intervention for excessive inflammation.


Assuntos
Brônquios/imunologia , Infecções por Burkholderia/imunologia , Burkholderia cenocepacia/imunologia , Coinfecção/imunologia , Inflamação/etiologia , Infecções Estafilocócicas/imunologia , Células Cultivadas , Células Epiteliais/imunologia , Humanos , Evasão da Resposta Imune , Interleucina-8/biossíntese , Sistema de Sinalização das MAP Quinases/fisiologia , NF-kappa B/fisiologia
4.
Mol Med Rep ; 20(4): 2990-3002, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31432147

RESUMO

Gene expression and DNA methylation levels affect the outcomes of patients with cancer. The present study aimed to establish a multigene risk model for predicting the outcomes of patients with cervical cancer (CerC) treated with or without radiotherapy. RNA sequencing training data with matched DNA methylation profiles were downloaded from The Cancer Genome Atlas database. Patients were divided into radiotherapy and non­radiotherapy groups according to the treatment strategy. Differently expressed and methylated genes between the two groups were identified, and 8 prognostic genes were identified using Cox regression analysis. The optimized risk model based on the 8­gene signature was defined using the Cox's proportional hazards model. Kaplan­Meier survival analysis indicated that patients with higher risk scores exhibited poorer survival compared with patients with lower risk scores (log­rank test, P=3.22x10­7). Validation using the GSE44001 gene set demonstrated that patients in the high­risk group exhibited a shorter survival time comprared with the low­risk group (log­rank test, P=3.01x10­3). The area under the receiver operating characteristic curve values for the training and validation sets were 0.951 and 0.929, respectively. Cox regression analyses indicated that recurrence and risk status were risk factors for poor outcomes in patients with CerC treated with or without radiotherapy. The present study defined that the 8­gene signature was an independent risk factor for the prognosis of patients with CerC. The 8­gene prognostic model had predictive power for CerC prognosis.


Assuntos
Regulação Neoplásica da Expressão Gênica , Interleucina-8/biossíntese , Modelos Biológicos , Proteínas de Neoplasias/biossíntese , Neoplasias do Colo do Útero , Adulto , Intervalo Livre de Doença , Feminino , Humanos , Interleucina-8/genética , Pessoa de Meia-Idade , Proteínas de Neoplasias/genética , Valor Preditivo dos Testes , Taxa de Sobrevida , Neoplasias do Colo do Útero/genética , Neoplasias do Colo do Útero/metabolismo , Neoplasias do Colo do Útero/mortalidade , Neoplasias do Colo do Útero/radioterapia
5.
Int J Mol Med ; 44(3): 949-959, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31257455

RESUMO

Pistacia weinmannifolia (PW) has been used in traditional Chinese medicine to treat headaches, dysentery, enteritis and influenza. However, PW has not been known for treating respiratory inflammatory diseases, including chronic obstructive pulmonary disease (COPD). The present in vitro analysis confirmed that PW root extract (PWRE) exerts anti­inflammatory effects in phorbol myristate acetate­ or tumor necrosis factor α (TNF­α)­stimulated human lung epithelial NCI­H292 cells by attenuating the expression of interleukin (IL)­8, IL­6 and Mucin A5 (MUC5AC), which are closely associated with the pulmonary inflammatory response in the pathogenesis of COPD. Thus, the aim of the present study was to evaluate the protective effect of PWRE on pulmonary inflammation induced by cigarette smoke (CS) and lipopolysaccharide (LPS). Treatment with PWRE significantly reduced the quantity of neutrophils and the levels of inflammatory molecules and toxic molecules, including tumor TNF­α, IL­6, IL­8, monocyte chemoattractant protein­1, neutrophil elastase and reactive oxygen species, in the bronchoalveolar lavage fluid of mice with CS­ and LPS­induced pulmonary inflammation. PWRE also attenuated the influx of inflammatory cells in the lung tissues. Furthermore, PWRE downregulated the activation of nuclear factor­κB and the expression of phosphodiesterase 4 in the lung tissues. Therefore, these findings suggest that PWRE may be a valuable adjuvant treatment for COPD.


Assuntos
Interleucina-8/biossíntese , Lipopolissacarídeos/efeitos adversos , NF-kappa B/metabolismo , Pistacia/química , Extratos Vegetais/farmacologia , Pneumonia Bacteriana/etiologia , Pneumonia Bacteriana/metabolismo , Fumaça/efeitos adversos , Animais , Linhagem Celular , Citocinas/metabolismo , Modelos Animais de Doenças , Mediadores da Inflamação/metabolismo , Lipopolissacarídeos/imunologia , Camundongos , Neutrófilos/imunologia , Neutrófilos/metabolismo , Extratos Vegetais/química , Fator de Necrose Tumoral alfa/metabolismo , Fator de Necrose Tumoral alfa/farmacologia
6.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 35(6): 498-504, 2019 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-31292053

RESUMO

Objective To observe the effect of selectively inhibiting STAT3 on the production of IL-8 and cell apoptosis of THP-1 cells by Stattic, and explore the underlying mechanism. Methods THP-1 cells were treated with different concentrations of Stattic ( 0, 1, 5, 10, 15, 20 µmol/L) for 0, 1, 3, 6, 12, 24 hours. Reverse transcription PCR or real-time PCR was performed to detect the mRNA expression of IL-8, IL-6, IL-1ß and tumor necrosis factor-α (TNF-α); ELISA was used to detect the protein expression of IL-8; flow cytometry was applied to evaluate the apoptosis of THP-1 cells; and Western blot analysis was performed to detect the phosphorylation of STAT3 and extracellular signal-regulated kinase (ERK). Reverse transcription PCR was used to test the effect of U0126 at different concentrations (0, 1, 5, 10 µmol/L) on the mRNA expression of IL-8 induced by Stattic in THP-1 cells. Results Stattic significantly up-regulated the mRNA and protein expression of IL-8 in THP-1 cells in a concentration range of 10~20 µmol/L, and induced cell apoptosis only at high concentration (15, 20 µmol/L). Treated with Stattic for 0, 1, 3, 6, 12, 24 hours, IL-8 mRNA was significantly up-regulated, and after 6 hours, the expression of IL-8 protein and apoptosis of THP-1 cells were up-regulated in a time-dependent manner. STAT3 phosphorylation was inhibited in a time- and dose-dependent manner by Stattic. ERK phosphorylation was induced by different concentrations of Stattic in a time-dependent manner. In addition, U0126, a selective inhibitor of ERK pathway, inhibited Stattic-induced IL-8 expression in a concentration-dependent manner. Conclusion Stattic, a selective STAT3 inhibitor, can induce the apoptosis and IL-8 production by activating ERK signaling pathway in THP-1 cells.


Assuntos
Apoptose , Óxidos S-Cíclicos/farmacologia , MAP Quinases Reguladas por Sinal Extracelular , Interleucina-8/biossíntese , Fator de Transcrição STAT3/antagonistas & inibidores , Humanos , Sistema de Sinalização das MAP Quinases , Fosforilação , Células THP-1
7.
Exp Mol Pathol ; 110: 104275, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31233733

RESUMO

Sulfur mustard (SM), a potent vesicating chemical warfare agent, and its analog nitrogen mustard (NM), are both strong bi-functional alkylating agents. Eyes, skin, and the respiratory system are the main targets of SM and NM exposure; however, ocular tissue is most sensitive, resulting in severe ocular injury. The mechanism of ocular injury from vesicating agents' exposure is not completely understood. To understand the injury mechanism from exposure to vesicating agents, NM has been previously employed in our toxicity studies on primary human corneal epithelial cells and ex vivo rabbit cornea organ culture model. In the current study, corneal toxicity from NM ocular exposure (1%) was analyzed for up to 28 days post-exposure in New Zealand White male rabbits to develop an acute corneal injury model. NM exposure led to conjunctival and eyelid swelling within a few hours after exposure, in addition to significant corneal opacity and ulceration. An increase in total corneal thickness and epithelial degradation was observed starting at day 3 post-NM exposure, which was maximal at day 14 post-exposure and did not resolve until 28 days post-exposure. There was an NM-induced increase in the number of blood vessels and inflammatory cells, and a decrease in keratocytes in the corneal stroma. NM exposure resulted in increased expression levels of cyclooxygenase-2, Interleukin-8, vascular endothelial growth factor and Matrix Metalloproteinase 9 indicating their involvement in NM-induced corneal injury. These clinical, biological, and molecular markers could be useful for the evaluation of acute corneal injury and to screen for therapies against NM- and SM-induced ocular injury.


Assuntos
Córnea/efeitos dos fármacos , Lesões da Córnea/metabolismo , Mecloretamina/toxicidade , Gás de Mostarda/toxicidade , Doença Aguda , Animais , Substâncias para a Guerra Química/toxicidade , Córnea/metabolismo , Córnea/patologia , Lesões da Córnea/induzido quimicamente , Ciclo-Oxigenase 2/biossíntese , Humanos , Imuno-Histoquímica , Interleucina-8/biossíntese , Masculino , Metaloproteinase 9 da Matriz/biossíntese , Coelhos , Fator A de Crescimento do Endotélio Vascular/biossíntese
8.
Environ Pollut ; 252(Pt B): 1216-1224, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31252119

RESUMO

The effects of methylamine on human health have been debated for several years, but the exact adverse outcomes and definite signaling cascades have not been elucidated yet. Herein, a NF-κB signal pathway, a positive regulator of inflammation was identified as the main pathway of methylamine exposure induced adverse effects in bronchial airway cells (16HBE) for the first time. The results indicated that methylamine could stimulate the overproduction of reactive oxygen species (ROS) in cytoplasm and mitochondria of 16HBE cells. Moreover, ROS accelerate the translocation and phosphorylation of NF-κB in nucleic and promote the expression of inflammatory, such as IL-8 and IL-6. As a result, methylamine was found to be increased ROS-mediated NF-κB activation in cells, leading to the production of inflammatory cytokine. Furthermore, the results also showed that methylamine could affect the expression of cytokines related genes, p53, STAT3, Bcl2, c-myc, Cyclin D, Hes1, Mcl-1, TGF-ß2. The breakdown of those cell proliferation and apoptosis related genes were leading to a common toxic mechanism of cell death. In summary, our work uncovers a mechanism by which methylamine can induce the formation of inflammation response and demonstrates potential inflammation and carcinogenesis in human airway cell upon the methylamine inhaled.


Assuntos
Poluentes Atmosféricos/toxicidade , Exposição Ambiental/efeitos adversos , Inflamação/patologia , Pneumopatias/induzido quimicamente , Metilaminas/toxicidade , Fator de Transcrição RelA/metabolismo , Apoptose/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Citocinas/metabolismo , Regulação da Expressão Gênica , Humanos , Interleucina-6/biossíntese , Interleucina-8/biossíntese , Mitocôndrias/metabolismo , Fosforilação , Transporte Proteico/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos
9.
Oncogene ; 38(28): 5566-5579, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31147602

RESUMO

Cancer-associated fibroblasts (CAFs), one of the major components of a tumour microenvironment, comprise heterogeneous populations involved in tumour progression. However, it remains obscure how CAF heterogeneity is governed by cancer cells. Here, we show that cancer extracellular vesicles (EVs) induce a series of chemokines in activated fibroblasts and contribute to the formation of the heterogeneity. In a xenograft model of diffuse-type gastric cancer, we showed two distinct fibroblast subpopulations with alpha-smooth muscle actin (α-SMA) expression or chemokine expression. MicroRNAs (miRNAs) profiling of the EVs and the transfection experiment suggested that several miRNAs played a role in the induction of chemokines such as CXCL1 and CXCL8 in fibroblasts, but not for the myofibroblastic differentiation. Clinically, aberrant activation of CXCL1 and CXCL8 in CAFs correlated with poorer survival in gastric cancer patients. Thus, this link between chemokine expression in CAFs and tumour progression may provide novel targets for anticancer therapy.


Assuntos
Fibroblastos Associados a Câncer/metabolismo , Quimiocina CXCL1/biossíntese , Vesículas Extracelulares/metabolismo , Interleucina-8/biossíntese , Neoplasias Gástricas/patologia , Células Estromais/patologia , Actinas/metabolismo , Animais , Linhagem Celular Tumoral , Xenoenxertos , Humanos , Camundongos , Neoplasias Gástricas/metabolismo , Microambiente Tumoral
10.
Molecules ; 24(9)2019 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-31072069

RESUMO

Nandina domestica (Berberidaceae) has been used in traditional medicine for the treatment of cough. This plant is distributed in Korea, Japan, China, and India This study aimed to investigate the anti-inflammatory phytochemicals obtained from the N. domestica fruits. We isolated a biflavonoid-type phytochemical, robustaflavone (R), from N. domestica fruits through bioactivity-guided fractionation based on its capacity to inhibit inflammation. The anti-inflammatory mechanism of R isolated from N. domestica has not yet been studied. In the present study, we evaluated the anti-inflammatory activities of R using lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. We have shown that R reduces the production of nitric oxide (NO), pro-inflammatory cytokine interleukin-1 beta (IL-1ß), and IL-6. Western blot analysis showed that R suppresses the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), and downregulates the expression of LPS-induced nuclear factor-kappa B (NF-κB) and the phosphorylation of extracellular-regulated kinases (pERK 1/2). Moreover, R inhibited IL-8 release in LPS-induced human colonic epithelial cells (HT-29). These results suggest that R could be a potential therapeutic candidate for inflammatory bowel disease (IBD).


Assuntos
Berberidaceae/química , Biflavonoides/isolamento & purificação , Biflavonoides/farmacologia , Regulação para Baixo , Mediadores da Inflamação/metabolismo , Animais , Biflavonoides/química , Morte Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Fracionamento Químico , Ciclo-Oxigenase 2/metabolismo , Regulação para Baixo/efeitos dos fármacos , Enterócitos/efeitos dos fármacos , Enterócitos/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Células HT29 , Humanos , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Interleucina-8/biossíntese , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , NF-kappa B/metabolismo , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase Tipo II/metabolismo , Fosforilação/efeitos dos fármacos , Compostos Fitoquímicos/química , Compostos Fitoquímicos/isolamento & purificação , Compostos Fitoquímicos/farmacologia , Extratos Vegetais/farmacologia , Células RAW 264.7
11.
PLoS Genet ; 15(4): e1008077, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30969964

RESUMO

The role of ribosomal protein S6 (rpS6) phosphorylation in mRNA translation remains poorly understood. Here, we reveal a potential role in modulating the translation rate of chemokine (C-X-C motif) ligand 8 (CXCL8 or Interleukin 8, IL8). We observed that more CXCL8 protein was being secreted from less CXCL8 mRNA in primary macrophages and macrophage-like HL-60 cells relative to other cell types. This correlated with an increase in CXCL8 polyribosome association, suggesting an increase in the rate of CXCL8 translation in macrophages. The cell type-specific expression levels were replicated by a CXCL8- UTR-reporter (Nanoluc reporter flanked by the 5' and 3' UTR of CXCL8). Mutations of the CXCL8-UTR-reporter revealed that cell type-specific expression required: 1) a 3' UTR of at least three hundred bases; and 2) an AU base content that exceeds fifty percent in the first hundred bases of the 3' UTR immediately after the stop codon, which we dub AU-rich proximal UTR sequences (APS). The 5' UTR of CXCL8 enhanced expression at the protein level and conferred cell type-specific expression when paired with a 3' UTR. A search for other APS-positive mRNAs uncovered TNF alpha induced protein 6 (TNFAIP6), another mRNA that was translationally upregulated in macrophages. The elevated translation of APS-positive mRNAs in macrophages coincided with elevated rpS6 S235/236 phosphorylation. Both were attenuated by the ERK1/2 signaling inhibitors, U0126 and AZD6244. In A549 cells, rpS6 S235/236 phosphorylation was induced by TAK1, Akt or PKA signaling. This enhanced the translation of the CXCL8-UTR-reporters. Thus, we propose that the induction of rpS6 S235/236 phosphorylation enhances the translation of mRNAs that contain APS motifs, such as CXCL8 and TNFAIP6. This may contribute to the role of macrophages as the primary producer of CXCL8, a cytokine that is essential for immune cell recruitment and activation.


Assuntos
Interleucina-8/biossíntese , Interleucina-8/genética , Proteína S6 Ribossômica/metabolismo , Células A549 , Elementos Ricos em Adenilato e Uridilato , Sequência de Bases , Moléculas de Adesão Celular/genética , Moléculas de Adesão Celular/metabolismo , Células Cultivadas , Fator de Iniciação 4E em Eucariotos/metabolismo , Células HL-60 , Humanos , Sistema de Sinalização das MAP Quinases , Macrófagos/imunologia , Macrófagos/metabolismo , Modelos Biológicos , Mutagênese , Fosforilação , Polirribossomos/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteína S6 Ribossômica/química , Proteína S6 Ribossômica/genética , Regiões não Traduzidas
12.
PLoS One ; 14(4): e0214847, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30947287

RESUMO

Due to their ability to preferentially induce cell death in tumor cells, while sparing healthy cells, TNF-related apoptosis-inducing ligand (TRAIL) and agonistic anti-TRAIL-R1 or anti-TRAIL-R2-specific antibodies are under clinical investigations for cancer-treatment. However, TRAIL-Rs may also induce signaling pathways, which result in malignant progression. TRAIL receptors are transcriptionally upregulated via wild-type p53 following radio- or chemotherapy. Nevertheless, the impact of p53 status on the expression and signaling of TRAIL-Rs is not fully understood. Therefore, we analyzed side by side apoptotic and non-apoptotic signaling induced by TRAIL or the agonistic TRAIL-R-specific antibodies Mapatumumab (anti-TRAIL-R1) and Lexatumumab (anti-TRAIL-R2) in the two isogenic colon carcinoma cell lines HCT116 p53+/+ and p53-/-. We found that HCT116 p53+/+ cells were significantly more sensitive to TRAIL-R-triggering than p53-/- cells. Similarly, A549 lung cancer cells expressing wild-type p53 were more sensitive to TRAIL-R-mediated cell death than their derivatives with knockdown of p53. Our data demonstrate that the contribution of p53 in regulating TRAIL-R-induced apoptosis does not correlate to the levels of TRAIL-Rs at the plasma membrane, but rather to p53-mediated upregulation of Bax, favouring the mitochondrial amplification loop. Consistently, stronger caspase-9 and caspase-3 activation as well as PARP-cleavage was observed following TRAIL-R-triggering in HCT116 p53+/+ compared to HCT116 p53-/- cells. Interestingly, HCT116 p53+/+ cells showed also a more potent activation of non-canonical TRAIL-R-induced signal transduction pathways like JNK, p38 and ERK1/ERK2 than p53-/- cells. Likewise, these cells induced IL-8 expression in response to TRAIL, Mapatumumab or Lexatumumab significantly stronger than p53-/- cells. We obtained similar results in A549 cells with or without p53-knockdown and in the two isogenic colon cancer cell lines RKO p53+/+ and p53-/-. In both cellular systems, we could clearly demonstrate the potentiating effects of p53 on TRAIL-R-mediated IL-8 induction. In conclusion, we found that wild-type p53 increases TRAIL-R-mediated apoptosis but simultaneously augments non-apoptotic signaling.


Assuntos
Apoptose/fisiologia , Neoplasias/metabolismo , Ligante Indutor de Apoptose Relacionado a TNF/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Células A549 , Membrana Celular/metabolismo , Técnicas de Silenciamento de Genes , Genes p53 , Células HCT116 , Humanos , Interleucina-8/biossíntese , Neoplasias/patologia , Receptor Ativador de Fator Nuclear kappa-B/metabolismo , Receptores do Ligante Indutor de Apoptose Relacionado a TNF/metabolismo , Transdução de Sinais , Proteína Supressora de Tumor p53/deficiência , Proteína X Associada a bcl-2/metabolismo
13.
Immunity ; 50(4): 1099-1114.e10, 2019 04 16.
Artigo em Inglês | MEDLINE | ID: mdl-30876876

RESUMO

Inflammatory bowel disease is a chronic, relapsing condition with two subtypes, Crohn's disease (CD) and ulcerative colitis (UC). Genome-wide association studies (GWASs) in UC implicate a FCGR2A variant that alters the binding affinity of the antibody receptor it encodes, FcγRIIA, for immunoglobulin G (IgG). Here, we aimed to understand the mechanisms whereby changes in FcγRIIA affinity would affect inflammation in an IgA-dominated organ. We found a profound induction of anti-commensal IgG and a concomitant increase in activating FcγR signaling in the colonic mucosa of UC patients. Commensal-IgG immune complexes engaged gut-resident FcγR-expressing macrophages, inducing NLRP3- and reactive-oxygen-species-dependent production of interleukin-1ß (IL-1ß) and neutrophil-recruiting chemokines. These responses were modulated by the FCGR2A genotype. In vivo manipulation of macrophage FcγR signal strength in a mouse model of UC determined the magnitude of intestinal inflammation and IL-1ß-dependent type 17 immunity. The identification of an important contribution of IgG-FcγR-dependent inflammation to UC has therapeutic implications.


Assuntos
Anticorpos Antibacterianos/imunologia , Colite Ulcerativa/imunologia , Microbioma Gastrointestinal/imunologia , Imunoglobulina G/imunologia , Interleucina-1beta/imunologia , Células Th17/imunologia , Animais , Colite/induzido quimicamente , Colite/imunologia , Colite/microbiologia , Colite/patologia , Colite Ulcerativa/microbiologia , Colite Ulcerativa/patologia , Sulfato de Dextrana/toxicidade , Regulação da Expressão Gênica , Genótipo , Humanos , Inflamação , Interleucina-8/biossíntese , Interleucina-8/genética , Mucosa Intestinal/imunologia , Mucosa Intestinal/microbiologia , Macrófagos/imunologia , Camundongos , Fagócitos/imunologia , RNA Mensageiro/biossíntese , Espécies Reativas de Oxigênio , Receptores de IgG/biossíntese , Receptores de IgG/genética , Receptores de IgG/imunologia
14.
Lett Appl Microbiol ; 68(6): 530-536, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30790328

RESUMO

We previously reported that oral administration of heat-killed Lactococcus lactis H61 improves certain human skin properties. For topical application of this strain, we reasoned that a bacterial cell extract obtained with an aqueous solvent could be readily formulated as a cosmetic ingredient. In the present study, we characterized the water extract from heat-killed H61. The extract had inhibitory activity for angiotensin-converting enzyme, which is known as suppression of inflammation of skin, and absorbed electromagnetic radiation in the UVB range. UVB-irradiated normal human epidermal keratinocytes (NHEKs) had lower viability than nonirradiated NHEKs. The NHEK survival rate was significantly higher in cells treated with the extract at 10 mg dried cells per ml prior to UVB exposure than in untreated cells or cells treated with lower extract concentrations. At this concentration, the extract also inhibited the production of interleukin-8 induced by UVB. The extract did not protect against hydrogen peroxide-induced cell damage. These data indicate that topical application of the H61 extract alleviates UVB damage and reduces inflammation in skin cells. The present study expands the potential application of strain H61 to its use as a cosmetic ingredient in addition to its use in the food industry. SIGNIFICANCE AND IMPACT OF THE STUDY: In our previous report, oral administration of heat-killed Lactococcus lactis H61 improved certain human skin properties. This study aimed exploring the potential topical use of this strain. The water extract derived from heat-killed cells with angiotensin-converting enzyme inhibitory activity, which is known as suppression of inflammation of skin, could protect normal human epidermal keratinocytes (NHEKs) from damage caused by UVB. Higher interleukin-8 production by UVB-exposed NHEKs than nontreated cells was suppressed by addition of the extract. The extract absorbed electromagnetic radiation in the UVB range. This extract could help in the maintenance of skin health by suppressing inflammation.


Assuntos
Queratinócitos/citologia , Lactococcus lactis/metabolismo , Fenômenos Fisiológicos da Pele/efeitos dos fármacos , Pele/microbiologia , Raios Ultravioleta/efeitos adversos , Animais , Cosméticos , Temperatura Alta , Humanos , Interleucina-8/biossíntese , Água/química
15.
Mol Cell Biochem ; 456(1-2): 135-144, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30715670

RESUMO

To identify PBMC-expressed genes significant for RA, and to ascertain their upstream regulatory factors, as well as downstream functional effects relevant to RA pathogenesis. We performed peripheral blood mononuclear cells (PBMCs) transcriptome-wide mRNA expression profiling in a case-control discovery sample. Differentially expressed genes (DEGs) were identified and validated in PBMCs in independent samples. We also generated genome-wide SNP genotyping data, and collected miRNA expression data and DNA methylation data from PBMCs of the discovery sample. Pearson correlation analyses were conducted to identify miRNAs/DNA methylations influencing DEG expression. Association analyses were conducted to identify expression-regulating SNPs. The key DEG, SAMD9, which was reported to function as a tumor suppressor gene, was assessed for its effects on T cell proliferation, apoptosis, and inflammatory cytokine expression. A total of 181 DEGs (Fold Change ≥ 2.0, Bonferroni adjusted p ≤ 0.05) were discovered in PBMCs. Four DEGs (SAMD9, CKLF, PARP9, and GUSB), upregulated with RA, were validated independently in PBMCs. Specifically, SAMD9 mRNA expression level was significantly upregulated in PHA-activated Jurkat T cells in vitro, and correlated with 8 miRNAs and associated with 22 SNPs in PBMCs in vivo. Knockdown of SAMD9 could transiently promote Jurkat T cell proliferation within 48 h and significantly induce TNF-α and IL-8 expression in T cells. SAMD9 expression is (epi-) genetically regulated, and significantly upregulated in PBMCs in RA patients and in activated T cells in vitro. SAMD9 might serve as a T cell activation marker but act as an anti-inflammatory factor.


Assuntos
Artrite Reumatoide , Proliferação de Células , Epigênese Genética , Polimorfismo de Nucleotídeo Único , Proteínas , Linfócitos T/metabolismo , Artrite Reumatoide/genética , Artrite Reumatoide/metabolismo , Artrite Reumatoide/patologia , Feminino , Estudo de Associação Genômica Ampla , Humanos , Interleucina-8/biossíntese , Interleucina-8/genética , Peptídeos e Proteínas de Sinalização Intracelular , Células Jurkat , Masculino , Proteínas/genética , Proteínas/metabolismo , Linfócitos T/patologia , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/genética
16.
Arch Biochem Biophys ; 664: 167-173, 2019 03 30.
Artigo em Inglês | MEDLINE | ID: mdl-30677406

RESUMO

Human airway trypsin-like protease (HAT) localizes at human bronchial epithelial cells (HBECs). HAT enhanced release of interleukin-8 (IL-8) from HBECs at 10-100 mU/mL and the enhanced release was almost completely abolished by 50 µM leupeptin, a serine protease inhibitor. Previous reports suggested that HAT displays its physiological functions via protease-activated receptor 2 (PAR2). In the present study, we examined the mechanism whereby HAT upregulates IL-8 synthesis in HBECs with a focus on PAR2. Northern blot analysis revealed that HAT enhanced IL-8 mRNA expression at concentrations of 10-100 mU/mL. PAR2 activating peptide (PAR2 AP) also enhanced IL-8 release and IL-8 mRNA expression in HBECs at 50-1,000 µM at similar levels as HAT. Knockdown of PAR2 mRNA by siRNA methods showed that PAR2 mRNA expression was significantly depressed in primary HBECs, and both HAT- and PAR2 AP-induced IL-8 mRNA elevation was significantly depressed in PAR2 siRNA-transfected HBECs. Additionally, HAT cleaved the PAR2 activating site (R36-S37 bond) of synthetic PAR2 N-terminal peptide. These results indicate that HAT stimulates IL-8 synthesis in airway epithelial cells via PAR2 and could help to amplify inflammation in chronic respiratory tract disease.


Assuntos
Brônquios/enzimologia , Interleucina-8/biossíntese , Receptor PAR-2/metabolismo , Serina Endopeptidases/metabolismo , Sequência de Aminoácidos , Brônquios/citologia , Células Cultivadas , Células Epiteliais/metabolismo , Humanos , Interleucina-8/genética , RNA Mensageiro/genética , RNA Interferente Pequeno/genética
17.
Environ Toxicol ; 34(4): 476-485, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30623574

RESUMO

Steroid-insensitive asthma-related airway inflammation is associated with the expression of epidermal growth factor receptor (EGFR) tyrosine kinase in asthmatic bronchial epithelium. Proinflammatory cytokines IL-6 and IL-8 are related to steroid-insensitive asthma. It is currently unknown how EGFR-tyrosine kinase inhibitors (EGFR-TKIs) affects house dust mite (HDM)-induced asthma in terms of inflammatory cytokines related to steroid-resistant asthma and further signaling pathway. Cytokine expressions and EGFR signaling pathway were performed by ELISA, reverse transcriptase PCR, real-time PCR, and Western blot in cell-line models. AMP-activated protein kinase (AMPK) pathway-related inhibitors were applied to confirm the association between EGFR-TKI and AMPK pathway. HDM induced IL-6 and IL-8 in a dose-dependent manner. Both Erlotinib (Tarceva) and Osimertinib (AZD-9291) reduced the levels of HDM-stimulated IL-6 and IL-8 levels in BEAS-2B cells. AZD-9291 was more effective than Erlotinib in inhibiting phospho-EGFR, and downstream phosphatidylinositol-3-kinase/protein kinase B (PI3K/AKT) and phopho-signal transducer and activator of transcription 3 (p-STAT3) pathway signaling. In addition, AMPK pathway-related inhibitor, Calcium-/calmodulin-dependent protein kinase kinase ß (CaMKKß) inhibitor, down-regulated IL-8, but EGFR-TKI had no effect on AMPK pathway. Our findings highlight EGFR-TKIs, Tarceva, and AZD-9291, attenuate HDM-induced inflammatory IL-6 and IL-8 cytokines via EGFR signaling axis pathway, but not AMPK signaling pathway.


Assuntos
Antígenos de Dermatophagoides/imunologia , Proteínas de Artrópodes/imunologia , Dermatophagoides pteronyssinus/imunologia , Células Epiteliais/efeitos dos fármacos , Receptores ErbB/antagonistas & inibidores , Interleucina-6/biossíntese , Interleucina-8/biossíntese , Acrilamidas/farmacologia , Compostos de Anilina/farmacologia , Animais , Asma/imunologia , Asma/prevenção & controle , Linhagem Celular , Relação Dose-Resposta a Droga , Células Epiteliais/imunologia , Receptores ErbB/metabolismo , Cloridrato de Erlotinib/farmacologia , Humanos , Mucosa Respiratória/efeitos dos fármacos , Mucosa Respiratória/imunologia , Transdução de Sinais
18.
PLoS One ; 14(1): e0210798, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30645630

RESUMO

Gut fungi may influence the course of Clostridium difficile infection either positively or negatively for the host. Fungi are not prominent in the mouse gut, and C. albicans, the major human gastrointestinal commensal yeast, is in low abundance or absent in mice. Bifidobacterium is one of the probiotics that may attenuate the severity of C. difficile infection. Inflammatory synergy between C. albicans and C. difficile, in gut, may provide a state that more closely resembles human infection and be more suitable for testing probiotic effects. We performed fecal mycobiota analysis and administered C. albicans at 1 day prior to C. difficile dosing. Fecal eukaryotic 18S rDNA analysis demonstrated the presence of Ascomycota, specifically, Candida spp., after oral antibiotics, despite negative fecal fungal culture. C. albicans administration enhanced the severity of the C. difficile infection model as determined by mortality rate, weight loss, gut leakage (FITC-dextran assay), and serum and intestinal tissue cytokines. This occurred without increased fecal C. difficile or bacteremia, in comparison with C. difficile gavage alone. Candida lysate with C. difficile increased IL-8 production from HT-29 and Caco-2 human intestinal epithelial cell-lines. Bifidobacterium attenuated the disease severity of the C. difficile plus Candida model. The reduced severity was associated with decreased Candida burdens in feces. In conclusion, gut C. albicans worsened C. difficile infection, possibly through exacerbation of inflammation. Hence, a mouse model of Clostridium difficile infection with C. albicans present in the gut may better model the human patient condition. Gut fungal mycobiome investigation in patients with C. difficile is warranted and may suggest therapeutic targets.


Assuntos
Bifidobacterium/fisiologia , Candida albicans/patogenicidade , Infecções por Clostridium/microbiologia , Administração Oral , Animais , Células CACO-2 , Infecções por Clostridium/terapia , Modelos Animais de Doenças , Microbioma Gastrointestinal/fisiologia , Células HT29 , Humanos , Interleucina-8/biossíntese , Mucosa Intestinal/microbiologia , Mucosa Intestinal/fisiopatologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Micobioma/fisiologia , Permeabilidade , Probióticos
19.
Gut ; 68(10): 1764-1773, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-30661053

RESUMO

OBJECTIVE: Our previous studies have identified CXCL8 as the crucial chemokine responsible for gastric cancer metastasis mediated by loss of RACK1. However, the regulatory effect of CXCL8 on immune surveillance in gastric cancer remains obscure. DESIGN: Flow cytometry analyses were performed to examine major source of CXCL8 and phenotypes of immune cells in fresh tumour tissues from 76 patients with gastric cancer. Real-time PCR was performed to analyse CXCL8 mRNA level in gastric cancer tissues. For immunohistochemical analyses, a total of 420 patients with gastric cancer undergoing curative resection were enrolled. In vitro culture of fresh tumour tissue was performed to evaluate the potential therapeutic effect of blocking CXCL8 pathway in gastric cancer. RESULTS: Increased level of CXCL8 indicates poor clinical outcome and tumour progression in patients with gastric cancer. In gastric cancer tissues, CXCL8 is predominantly secreted by macrophages and colony stimulating factor 2 (CSF-2) facilitates macrophage-derived CXCL8 secretion. High level of CXCL8 is associated with decreased CD8+ T cells infiltration and Ki67+ CD8+ T cells proportion. Moreover, CXCL8 also inhibits CD8+ T cells function by inducing the expression of PD-L1 on macrophages. Finally, we show that a small-molecule CXCR2 inhibitor, reparixin, drives the decreased programmed death-ligand 1 (PD-L1+) macrophages and promotes antitumour immunity. Accordingly, high levels of CXCL8+ macrophages are positively correlated with poor prognosis in patients with gastric cancer. CONCLUSIONS: CXCL8 is predominantly secreted by macrophages and contributes to the immunosuppressive microenvironment by inducing PD-L1+ macrophages in gastric cancer. CXCL8 inhibitors may drive antitumour response, providing potential therapeutic effects for patients with gastric cancer.


Assuntos
Antígeno B7-H1/genética , Regulação Neoplásica da Expressão Gênica , Interleucina-8/genética , Linfócitos do Interstício Tumoral/metabolismo , Neoplasias Gástricas/genética , Antígeno B7-H1/biossíntese , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , DNA de Neoplasias/genética , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Humanos , Imuno-Histoquímica , Interleucina-8/biossíntese , Linfócitos do Interstício Tumoral/imunologia , Linfócitos do Interstício Tumoral/patologia , Masculino , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Neoplasias Gástricas/imunologia , Neoplasias Gástricas/patologia , Células Tumorais Cultivadas
20.
Biochem Cell Biol ; 97(2): 109-117, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30110560

RESUMO

During the pathogenetic process of varied kidney diseases, renal tubules are the major sites in response to detrimental insults, including pro-inflammatory stimuli. MicroRNA-204-5p (miR-204-5p) can be detected in the renal tubular epithelial cells in the normal kidney; its expression, however, is downregulated in the kidney with pathological changes. This study aimed to investigate the role of miR-204-5p in interleukin 6 (IL6) mediated inflammatory response and chemokine production in HK-2 renal tubular cells. In HK-2 cells, the expression of miR-204-5p was downregulated in response to exogenous pro-inflammatory stimulus, tumor necrosis factor α (TNFα), or IL1ß, while that of IL6 receptor α (IL6R) was upregulated. Dual-luciferase results confirmed that miRNA-204-5p directly targeted IL6R. In addition to suppressing IL6R expression, miRNA-204-5p agomir also inhibited the phosphorylation of signal transducer and activator of transcription 3 (STAT3) in HK-2 cells exposed to exogenous IL6. Further, miRNA-204-5p suppressed the overproduction of pro-inflammatory mediators (cyclooxygenase 2 and prostaglandin E2) and chemokines (C-C motif chemokine ligand 2 and C-X-C motif chemokine ligand 8). The anti-inflammatory effects of miRNA-204-5p were attenuated when IL6R was reexpressed in HK-2 cells. Collectively, our study reveals that miR-204-5p inhibits the inflammation and chemokine generation in renal tubular epithelial cells by modulating the IL6/IL6R axis.


Assuntos
Quimiocina CCL2/biossíntese , Células Epiteliais/metabolismo , Interleucina-6/metabolismo , Interleucina-8/biossíntese , Túbulos Renais Proximais/metabolismo , MicroRNAs/metabolismo , Receptores de Interleucina-6/metabolismo , Linhagem Celular , Quimiocina CCL2/genética , Células Epiteliais/patologia , Humanos , Inflamação/genética , Inflamação/metabolismo , Inflamação/patologia , Interleucina-6/genética , Interleucina-8/genética , Túbulos Renais Proximais/patologia , MicroRNAs/genética , Receptores de Interleucina-6/genética
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