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1.
Clin Rheumatol ; 2019 Nov 23.
Artigo em Inglês | MEDLINE | ID: mdl-31760539

RESUMO

In this study, we aimed to explore the expression levels of JAK2 and PTPRC in peripheral blood mononuclear cells (PBMCs) from SLE patients and controls, detect the effects of SLE activity on genes mRNA expression, and find the association between genes mRNA expression and clinical manifestations of patients. We performed quantitative real-time PCR (qRT-PCR) to test differences in the expression levels of JAK2 and PTPRC in PBMCs extracted from 135 patients with SLE and 130 healthy controls. Furthermore, we detected the regulatory effect of SNPs on gene expression by expression quantitative trait loci (eQTL). We also tested whether the genes mRNA expression was affected with the SLE activity and analyzed the relationship between genes mRNA expression and clinical manifestations of patients. The mRNA expression levels of JAK2 in SLE patients were significantly higher than those in healthy controls (P = 0.005), and PTPRC mRNA expression levels were significantly decreased (P < 0.001). However, no other statistical significance was detected. We found that the elevated JAK2 mRNA expression and the decreased PTPRC mRNA expression may play suggestive roles in the pathogenesis of SLE.Key Points• The JAK2 mRNA expression levels in SLE patients were significantly higher than those in healthy controls.• The PTPRC mRNA expression levels in SLE were decreased.• JAK2 and PTPRC mRNA expression may play suggestive roles in the pathogenesis of SLE.

2.
Mol Biol Rep ; 46(1): 1189-1197, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30632069

RESUMO

Interleukin (IL) 28 receptor α (IL28RA) is a well-known candidate for psoriasis susceptibility based on previous genome-wide association study (GWAS) analysis. However, the function of IL28RA in psoriasis has not been elucidated. In the present study, the expression of IL28RA was significantly decreased in lesional tissues from patients with plaque psoriasis when compared with the expression observed in adjacent non-lesional tissues. In vitro studies further demonstrated that in the presence of IL-29, HaCaT keratinocytes with IL28RA knockdown exhibited a faster rate of proliferation than control cells, and an enhanced ratio of cells in the S and G2/M phase. By contrast, IL28RA overexpression inhibited the proliferation of HaCaT keratinocytes and caused cell cycle arrest at the G0/G1 phases. Western blot analysis revealed that knockdown of IL28RA upregulated cyclinB1 expression and downregulated cyclinE expression; the opposite results were observed in the IL28RA-overexpressing HaCaT cells. Finally, a mechanistic study revealed that IL28RA functions through the activation of the Janus kinase-signal transducer and activator of transcription signaling pathway to exert its anti-proliferative effect. These results suggested that weak expression of IL28RA may contribute to the pathogenesis of psoriasis and that IL28RA may be an effective drug target for the treatment of psoriasis. However, further in vivo studies are required.


Assuntos
Queratinócitos/fisiologia , Psoríase/metabolismo , Receptores de Citocinas/metabolismo , Adulto , Ciclo Celular/fisiologia , Linhagem Celular , Proliferação de Células/fisiologia , Regulação para Baixo , Epiderme/metabolismo , Estudo de Associação Genômica Ampla , Humanos , Interferons , Interleucinas/genética , Interleucinas/fisiologia , Queratinócitos/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Psoríase/genética , Receptores de Citocinas/fisiologia , Transdução de Sinais , Ativação Transcricional , Regulação para Cima
3.
J Immunol Res ; 2018: 3651743, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30013990

RESUMO

Cationic protein is a cytotoxic protein secreted by eosinophils and takes part in the damage of airway epithelium in asthma. Poly-L-arginine (PLA), a synthetic cationic protein, is widely used to mimic the biological function of the natural cationic protein in vitro. Previous studies demonstrated the damage of the airway epithelial cells by cationic protein, but the molecular mechanism is unclear. The purpose of this study aimed at exploring whether PLA could induce apoptosis of human airway epithelial cells (NCI-H292) and the underlying mechanism. Methods. The morphology of apoptotic cells was observed by transmission electron microscopy. The rate of apoptosis was analyzed by flow cytometry (FCM). The expressions of the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2), Bcl-2/Bax, and cleaved caspase-3 were assessed by western blot. Results. PLA can induce apoptosis in NCI-H292 cells in a concentration-dependent manner. Moreover, the phosphorylation of the ERK1/2 and the unbalance of Bcl2/Bax, as well as the activation of caspase-3, were involved in the PLA-induced apoptosis. Conclusions. PLA can induce the apoptosis in NCI-H292 cells, and this process at least involved the ERK1/2 and mitochondrial pathway. The results could have some indications in revealing the apoptotic damage of the airway epithelial cells. Besides, inhibition of cationic protein-induced apoptotic death in airway epithelial cells could be considered as a potential target of anti-injury or antiremodeling in asthmatics.


Assuntos
Apoptose/efeitos dos fármacos , Carcinoma Mucoepidermoide/tratamento farmacológico , Neoplasias Pulmonares/tratamento farmacológico , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Peptídeos/farmacologia , Células Epiteliais Alveolares/efeitos dos fármacos , Células Epiteliais Alveolares/enzimologia , Células Epiteliais Alveolares/metabolismo , Carcinoma Mucoepidermoide/enzimologia , Carcinoma Mucoepidermoide/metabolismo , Carcinoma Mucoepidermoide/patologia , Caspase 3/metabolismo , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Humanos , Neoplasias Pulmonares/enzimologia , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Proteína Quinase 1 Ativada por Mitógeno/antagonistas & inibidores , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/antagonistas & inibidores , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Fosforilação , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo
4.
Planta ; 247(6): 1307-1321, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29504038

RESUMO

MAIN CONCLUSION: Transcriptome analysis was carried out for wheat seedlings and spikes from hybrid Jingmai 8 and both inbred lines to unravel mechanisms underlying heterosis. Heterosis, known as one of the most successful strategies for increasing crop yield, has been widely exploited in plant breeding systems. Despite its great importance, the molecular mechanism underlying heterosis remains elusive. In the present study, RNA sequencing (RNA-seq) was performed on the seedling and spike tissues of the wheat (Triticum aestivum) hybrid Jingmai 8 (JM8) and its homozygous parents to unravel the underlying mechanisms of wheat heterosis. In total, 1686 and 2334 genes were identified as differentially expressed genes (DEGs) between the hybrid and the two inbred lines in seedling and spike tissues, respectively. Gene Ontology analysis revealed that DEGs from seedling tissues were significantly enriched in processes involved in photosynthesis and carbon fixation, and the majority of these DEGs expressed at a higher level in JM8 compared to both inbred lines. In addition, cell wall biogenesis and protein biosynthesis-related pathways were also significantly represented. These results confirmed that a combination of different pathways could contribute to heterosis. The DEGs between the hybrid and the two inbred progenitors from the spike tissues were significantly enriched in biological processes related to transcription, RNA biosynthesis and molecular function categories related to transcription factor activities. Furthermore, transcription factors such as NAC, ERF, and TIF-IIA were highly expressed in the hybrid JM8. These results may provide valuable insights into the molecular mechanisms underlying wheat heterosis.


Assuntos
Regulação da Expressão Gênica de Plantas , Vigor Híbrido/genética , Transcriptoma , Triticum/genética , Perfilação da Expressão Gênica , Ontologia Genética , Endogamia , Inflorescência/genética , Inflorescência/fisiologia , Fotossíntese , Plântula/genética , Plântula/fisiologia , Análise de Sequência de RNA , Triticum/fisiologia
5.
Mol Med Rep ; 16(4): 4811-4816, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28765912

RESUMO

Mutation of genes encoding the enzymes of the mevalonate pathway cause a variety of diseases, including skin disorders. Mutation of four genes in this pathway, including mevalonate kinase, phosphomevalonate kinase, mevalonate diphosphate decarboxylase and farnesyl diphosphate synthase, have demonstrated to be responsible for porokeratosis (PK). However, the pathogenesis of PK remains unclear. In the present study, specific enzyme inhibitors of the mevalonate pathway, including pravastatin (PRA), alendronate (ALD), farnesyl transferase inhibitor (FTI­277) and geranylgeranyl transferase inhibitor (GGTI­298), were used to investigate the effect on differentiation of keratinocytes (KCs). Western blotting demonstrated that PRA, ALD, FTI­277 or GGTI­298 alone, or in combination, inhibited the expression level of calcium­induced differentiation maker involucrin (INV) in KCs. ALD and PRA induced greater inhibition of INV compared with FTI­277 and GGTI­298 treatment. These inhibitors additionally influenced the expression levels of keratin1. Mechanistic studies revealed that treatment of cells with inhibitors decreased the expression levels of p53 and Notch1, and regulated activation of the mitogen activated protein kinase and phosphoinositide­3­kinase/protein kinase B signaling pathways. The results of the present study suggested that regulation of the mevalonate pathway may be necessary for differentiation of KCs, and the pathogenesis of disseminated superficial actinic PK.


Assuntos
Cálcio/metabolismo , Diferenciação Celular , Queratinócitos/citologia , Queratinócitos/metabolismo , Redes e Vias Metabólicas , Ácido Mevalônico/metabolismo , Biomarcadores , Diferenciação Celular/genética , Expressão Gênica , Regulação da Expressão Gênica , Humanos , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptor Notch1/genética , Receptor Notch1/metabolismo , Transdução de Sinais , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo
6.
Exp Dermatol ; 25(7): 544-7, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-26914593

RESUMO

Interleukin (IL)-15 is an important proinflammatory cytokine that can protect epidermal keratinocytes (KCs) from ultraviolet-induced apoptosis and plays a role in the pathogenesis of psoriasis. However, the impact of IL-15 on KC differentiation remains unknown. In this study, isolated human primary epidermal KCs were treated with various concentrations of IL-15 for different times, and the expression of differentiation markers (keratin 1, involucrin and loricrin) and p53 as well as the activation of ERK, AKT and Notch induced by IL-15 in the absence or presence of Ca(2+) was detected by real-time PCR and Western blot. The results showed that stimulation with Ca(2+) alone increased the expression of KC differentiation markers and p53 and promoted the activation of Notch1. Pretreatment with IL-15 resulted in a decrease in the Ca(2+) -induced expression of KC differentiation markers and p53. Additionally, Ca(2+) continually inhibited the phosphorylation of ERK1/2 and activated AKT, and IL-15 reduced the effect of Ca(2+) on ERK1/2 and AKT. FR180204, a specific inhibitor of ERK1/2 phosphorylation, slightly attenuated the effect of Ca(2+) on the expression of differentiation markers and p53 and the activation of Notch1. In contrast, MK-2206, an inhibitor of pAKT, strongly blocked the expression of the differentiation markers and p53 and the activation of Notch1. An anti-IL-15 antibody neutralized the effect of IL-15 on KC differentiation. These results indicate that IL-15 inhibits the Ca(2+) -induced differentiation of KCs, mainly via the attenuation of Ca(2+) -stimulated PI3K-AKT signalling.


Assuntos
Cálcio/metabolismo , Interleucina-15/metabolismo , Queratinócitos/metabolismo , Diferenciação Celular , Expressão Gênica , Humanos , Queratinócitos/citologia , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação , Cultura Primária de Células , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptor Notch1/metabolismo
7.
Mol Med Rep ; 13(2): 1807-12, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26718740

RESUMO

Toll-like receptors (TLRs) are critical in the induction of the immune response in tumor development. TLR7 has previously been demonstrated to be associated with the development of pancreatic cancer, and the release of cytokines and chemokines from other types of cancer cell; however, the specific expression induced by TLR7 agonists in pancreatic cancer cells remains to be elucidated. The present study aimed to investigate the effects of the TLR7 agonist, gardiquimod, on ERK1/2 signaling pathway, and on the expression of genes involved in the pathogenesis of cancer, including phosphatase and tensin homolog deleted on chromosome 10 (PTEN), p53, type Ⅲ interferon (IFN-λ1), vascular endothelial growth factor (VEGF), matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1). The results demonstrated that activation of TLR7 upregulated the expression levels of certain genes to varying degrees; the expression levels of IFN-λ1 and MMP-9 were increased by ~3 fold, whereas other genes (p53, PTEN, TIMP-1) were upregulated by ~2 fold, and VEGF was marginally upregulated after 10 min. Furthermore, gardiquimod increased the expression levels of phosphorylated-extracellular signal-regulated kinase (ERK)1/2. In addition, PD98059, a specific inhibitor of ERK phosphorylation, inhibited the ability of gardiquimod to activate ERK1/2; consequently weakening the effect of gardiquimod on gene regulation. These findings indicated that the effect of TLR7 agonists, including gardiquimod, on gene expression in BxPC-3 pancreatic cancer cells was partly associated with the mitogen-activated protein kinase-ERK1/2 signaling pathway.


Assuntos
Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Interleucinas/genética , Metaloproteinase 9 da Matriz/genética , PTEN Fosfo-Hidrolase/genética , Neoplasias Pancreáticas/genética , Inibidor Tecidual de Metaloproteinase-1/genética , Receptor 7 Toll-Like/metabolismo , Proteína Supressora de Tumor p53/genética , Fator A de Crescimento do Endotélio Vascular/genética , Aminoquinolinas/farmacologia , Linhagem Celular Tumoral , Ativação Enzimática/efeitos dos fármacos , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Flavonoides/farmacologia , Humanos , Imidazóis/farmacologia , Interferons , Interleucinas/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , PTEN Fosfo-Hidrolase/metabolismo , Fosforilação/efeitos dos fármacos , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Regulação para Cima/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/metabolismo
8.
J Dermatol ; 43(1): 99-102, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26134586

RESUMO

We performed a gene expression study to explore whether expression levels of LBH contribute to the pathogenesis of systemic lupus erythematosus (SLE) and are associated with the genome-wide association study-identified SNP (rs7579944 and rs906868) and the SLE Disease Activity Index (SLEDAI). Fluorescent quantitative reverse transcription polymerase chain reaction was used to examine the expression of LBH mRNA in peripheral blood mononuclear cells (PBMC) from 62 SLE patients and 69 controls. The expression levels of LBH mRNA in patients with SLE were significantly decreased compared with those in normal controls (P < 0.001). No significant differences were found between LBH mRNA expression levels and SLEDAI scores, SNP rs7579944 and rs906868. This study suggests that decreased expression of LBH mRNA may be correlated with the pathogenesis of SLE.


Assuntos
Lúpus Eritematoso Sistêmico/genética , RNA Mensageiro/genética , Transativadores/genética , Adolescente , Adulto , Idoso , Estudos de Casos e Controles , Regulação para Baixo , Feminino , Humanos , Leucócitos Mononucleares/metabolismo , Lúpus Eritematoso Sistêmico/sangue , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , RNA Mensageiro/sangue , Adulto Jovem
9.
Inflammation ; 39(1): 47-53, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26246181

RESUMO

Major basic protein (MBP) derived from activated eosinophil can exacerbate atopic asthma induced by lipopolysaccharide (LPS). The pharmacological function of MBP can be mimicked by poly-L-arginine (PLA), however, the potential signaling mechanisms of LPS-PLA-induced release of the inflammatory cytokines interleukin (IL)-6 and IL-8 remain unclear. In the present study, airway epithelia NCI-H292 cell lines were treated with LPS and/or PLA. We found that the expression levels of IL-6 and IL-8 induced by LPS-PLA were increased significantly compared with that in untreated cells. Meanwhile, the phosphorylation of p38 MAPK and ERK1/2 was also up-regulated dramatically by LPS-PLA, but this increase could be blocked by specific inhibitor. Importantly, blocking the phosphorylation of p38 MAPK and ERK1/2 reduced the expression levels of IL-6 and IL-8 as well. Collectively, LPS-PLA-induced release of IL-6 and IL-8 from NCI-H292 cells may be due to the synergistic activation of p38 MAPK and ERK1/2 signaling transduction pathways.


Assuntos
Asma/patologia , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Lipopolissacarídeos/farmacologia , Peptídeos/farmacologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Linhagem Celular Tumoral , Ativação Enzimática/efeitos dos fármacos , Proteína Básica Maior de Eosinófilos/farmacologia , Humanos , Fosforilação/efeitos dos fármacos , Mucosa Respiratória/citologia , Mucosa Respiratória/fisiologia
10.
J Immunol Res ; 2016: 3421060, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-28116315

RESUMO

Objective. Herein, we aimed to study the mechanism whereby poly-L-arginine (PLA) and lipopolysaccharide (LPS) can synergistically induce the release of interleukin-6 (IL-6) and IL-8 in NCI-H292 cells. Methods. NCI-H292 cells were divided into control, PLA, LPS, and PLA+LPS groups. At various time points, the phosphorylation of JNK in each group was measured by western blotting. Additionally, the productions of IL-6 and IL-8 were assessed using an enzyme-linked immunosorbent assay (ELISA). The effects of SP600125, an inhibitor of the JNK pathway, on the increase of p-JNK, IL-6, and IL-8 were also studied. Results. Our results showed that either PLA or LPS treatment alone can significantly increase the phosphorylation level of JNK in NCI-H292 cells. Of interest was the combined use of PLA and LPS that has a synergistic effect on the phosphorylation of JNK, as well as synergistically inducing the release of IL-6 and IL-8 in NCI-H292 cells. Furthermore, SP600125 significantly inhibited the activation of JNK signal, as well as reducing the productions of IL-6 and IL-8 in response to PLA+LPS stimulation. Conclusions. The JNK signaling pathway contributes to the release of IL-6 and IL-8, which is stimulated by the synergistic actions of PLA+LPS in NCI-H292 cells.


Assuntos
Interleucina-6/metabolismo , Interleucina-8/metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Lipopolissacarídeos/farmacologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Peptídeos/farmacologia , Antracenos/farmacologia , Asma/patologia , Linhagem Celular Tumoral , Sinergismo Farmacológico , Ensaio de Imunoadsorção Enzimática , Humanos , Interleucina-6/biossíntese , Interleucina-8/biossíntese , Proteínas Quinases JNK Ativadas por Mitógeno/antagonistas & inibidores , Fosforilação/efeitos dos fármacos
11.
Mol Med Rep ; 12(4): 6079-85, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26238718

RESUMO

Pancreatic cancer is one of the most malignant types of tumor and has a poor prognosis. Toll­like receptor 7 (TLR7) has been found to be present and have different roles in different types of cancer cells. In the present study, the roles of TLR7 in BxPC­3 cells, a human pancreatic adenocarcinoma cell line, were investigated. The cells were treated with gardiquimod, an agonist of TLR7, following which the properties of the cells, including proliferation, migration, cell cycle and apoptosis, were analyzed. It was revealed that activation of TLR7 by gardiquimod inhibited cell proliferation and migration, and induced apoptosis of the cells. In addition, gardiquimod downregulated the expression levels of cyclin B1, cyclin E and B­cell lymphoma 2, while upregulating the expression of B­cell­associated X protein. These results suggested that the activation of TLR7 suppresses the progression of pancreatic cancer.


Assuntos
Apoptose/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Neoplasias Pancreáticas/patologia , Receptor 7 Toll-Like/metabolismo , Aminoquinolinas/farmacologia , Linfócitos B/metabolismo , Linhagem Celular Tumoral , Ciclina E/genética , Ciclina E/metabolismo , Relação Dose-Resposta a Droga , Regulação para Baixo , Humanos , Imidazóis/farmacologia , Proteínas Oncogênicas/genética , Proteínas Oncogênicas/metabolismo , Receptor 7 Toll-Like/agonistas , Receptor 7 Toll-Like/genética , Regulação para Cima , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo , beta Catenina/genética , beta Catenina/metabolismo
13.
Clin Rheumatol ; 34(10): 1807-11, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25911176

RESUMO

Systemic lupus erythematosus (SLE) is a systemic autoimmune and inflammatory disease with a strong genetic contribution and characterized by kinds of immune reactions. Our previous genome-wide association studies have identified IL-28RA as a susceptibility gene for SLE. In this study, we performed a quantitative reverse transcription polymerase chain reaction (RT-PCR) in 62 patients with SLE and 69 controls to investigate the different expression levels of IL-28RA in peripheral blood mononuclear cells (PBMCs) from SLE patients and healthy controls and the association between IL-28RA expression and systemic lupus erythematosus disease activity index (SLEDAI) or the variant of the single-nucleotide polymorphism (SNP) rs4649203. The expression levels of IL-28RA messenger RNA (mRNA) in SLE patients were significantly increased compared with those of healthy controls. In addition, there were also significant differences in the expression levels of IL-28RA between active (SLEDAI ≥ 6) or inactive (SLEDAI < 6) SLE groups and healthy controls. However, no correlation was observed between IL-28RA mRNA expression level and SLEDAI. There was no association between the variant of the SNP rs4649203 and IL-28RA mRNA expression levels neither. These results indicated that expression of IL-28RA mRNA may be correlated with the pathogenesis of SLE.


Assuntos
Leucócitos Mononucleares/citologia , Lúpus Eritematoso Sistêmico/sangue , Receptores de Citocinas/sangue , Adulto , Alelos , Estudos de Casos e Controles , Feminino , Variação Genética , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa
14.
Zookeys ; (438): 45-55, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25197218

RESUMO

Two new species of the stonefly genus Neoperla, N. nigromarginata sp. n. and N. similiflavescens sp. n., are described from Dabie Mountains of Central China in the Liankangshan National Nature Reserve. The new species are compared with related congeners.

15.
J Immunol ; 192(9): 4192-201, 2014 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-24670802

RESUMO

Clinical trials have shown that BAFF inhibitors do not reduce memory B cell levels but can reduce the number of mature B cells. It remains uncertain whether BAFF affects memory-maintaining cytokines such as IL-15. We found that BAFF suppressed IL-15 expression in B cells from lupus-like or experimental allergic encephalomyelitis mice. When BAFF was blocked with atacicept-IgG, IL-15 expression was upregulated in lupus-like or experimental allergic encephalomyelitis mice. Finally, we showed that BAFF suppressed IL-15 expression in transitional 2 B cells by reducing Foxo1 expression and inducing Foxo1 phosphorylation. This study suggests that BAFF suppresses IL-15 expression in autoimmune diseases, and this opens up the possible opportunity for the clinical application of BAFF- and IL-15-specific therapeutic agents.


Assuntos
Doenças Autoimunes/imunologia , Fator Ativador de Células B/imunologia , Linfócitos B/imunologia , Interleucina-15/biossíntese , Animais , Doenças Autoimunes/metabolismo , Fator Ativador de Células B/metabolismo , Linfócitos B/metabolismo , Western Blotting , Encefalomielite Autoimune Experimental/imunologia , Encefalomielite Autoimune Experimental/metabolismo , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Humanos , Interleucina-15/imunologia , Lúpus Eritematoso Sistêmico/sangue , Lúpus Eritematoso Sistêmico/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Esclerose Múltipla/sangue , Esclerose Múltipla/imunologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa
16.
Neurosci Lett ; 562: 54-9, 2014 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-24462842

RESUMO

Astrocytes undergo de-differentiation and become activated during a response to injury. Several studies have found that reactive astrocytes re-express markers, such as Nestin, which are normally expressed in neural stem cells. It was recently shown that the epidermal growth factor receptor (EGFR) is up-regulated in astrocytes after injury and promotes reactive astrocyte transformation. However, the signaling pathways involved in this process have not been elucidated. In the present study, we showed that Nestin was strongly expressed in reactive astrocytes. Furthermore, as shown by immunoblot analyses, epidermal growth factor (EGF) regulated Nestin expression through EGFR activation. Inhibition of the PLCγ, PI3K, ERK, p38, and JNK pathways did not affect Nestin expression in reactive astrocytes. However, treatment with a Raf-1 inhibitor inhibited Nestin expression in a concentration-dependent manner. Taken together, the signaling analyses revealed that EGF induced and regulated Nestin expression through activation of the Ras-Raf--ERK signaling pathway. This is the first study to show that Nestin expression is regulated by an extracellular signaling molecule in reactive astrocytes.


Assuntos
Astrócitos/efeitos dos fármacos , Fator de Crescimento Epidérmico/farmacologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Nestina/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Astrócitos/metabolismo , Fator de Crescimento Epidérmico/administração & dosagem , Receptores ErbB/efeitos dos fármacos , Receptores ErbB/metabolismo , Proteínas de Filamentos Intermediários/metabolismo , Nestina/metabolismo , Fosforilação/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Quinases raf/metabolismo , Proteínas ras/metabolismo
17.
Mol Cell Biochem ; 389(1-2): 43-9, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24347177

RESUMO

Toll-like receptors (TLRs) play important roles in activation of immunoreaction and tumor development. Toll-like receptor 7 (TLR7), one of the TLRs binding with single-stranded RNA, activates intracellular pathways and stimulates the release of proinflammatory cytokines, chemokines. In this study, we investigated the impact of the TLR7-signaling pathway on the expression of vascular endothelial growth factor (VEGF), matrix metalloproteinase 2 (MMP2), tissue inhibitor of metalloproteinase 1 (TIMP1), interleukin 6 (IL-6), and interleukin 15 (IL-15), which have been testified to refer to the immunomodulating and tumor progression. We confirmed that the TLR7 was expressed by Hela cells, despite the abundance was weak. Gardiquimod, one of the TLR7 ligands, can promote these five genes expression in varying degrees. After stimulating with gardiquimod, the expression of the IL-15V1, 3 increased about 4.5 times on RNA level, the other expression was only up-regulated about 2 times. We also discovered that gardiquimod could activate the MAPK/ERK- and PI3K/AKT-signaling pathways, and the specific inhibitors studies indicate that, the effect of gardiquimod on these genes expression is mainly or partially dependent on the activation of these two signaling pathways. To sum up, the activation of TLR7 signaling pathway may modulate some genes expression in Hela cells and may contribute to the pathogenesis of the cervical cancer.


Assuntos
Expressão Gênica/genética , Interleucina-15/genética , Interleucina-6/genética , Metaloproteinase 2 da Matriz/genética , Inibidor Tecidual de Metaloproteinase-1/genética , Receptor 7 Toll-Like/genética , Fator A de Crescimento do Endotélio Vascular/genética , Linhagem Celular Tumoral , Células HeLa , Humanos , Transdução de Sinais/genética
18.
Mol Biol Rep ; 40(11): 6363-9, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24057248

RESUMO

Toll-like receptor 7 (TLR7) is an important member in pattern recognition receptors families. TLR7 signal pathway is involved in the physiological process in many type cells, but the impact of TRL7 on differentiation in the human keratinocytes is still unknown. In this study, we investigated the expression of TLR7 in keratinocytes, and the effect of TLR7 agonist gardiquimod on the expression of calcium (Ca(2+))-induced keratinocytes differentiation markers in HaCaT cells. Immunohistochemistry and western-blotting analysis showed that TLR7 is expressed in basal keratinocytes of normal skin and in the human keratinocyte cell line HaCaT, but not expressed in the keratinocytes of psoriasis lesions. Pretreatment with gardiquimod could down-regulate Ca(2+)-induced differentiation marker expression and activate Raf-MEK-ERK and PI3K-AKT signal pathways in HaCaT cells. However, specific inhibitors studies showed that the down-regulation of the differentiation markers expression by gardiquimod was not dependent on the activation of these two pathways. TLR7 may play an important role in the pathogenesis of psoriasis through regulating the differentiation of the keratinocytes, and will give a new insight into the psoriasis.


Assuntos
Aminoquinolinas/farmacologia , Antígenos de Diferenciação/genética , Cálcio/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Imidazóis/farmacologia , Linhagem Celular , Humanos , Queratina-1/genética , Queratina-1/metabolismo , Queratinócitos/efeitos dos fármacos , Queratinócitos/metabolismo , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Precursores de Proteínas/genética , Precursores de Proteínas/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Psoríase/genética , Psoríase/metabolismo , Transdução de Sinais/efeitos dos fármacos , Receptor 7 Toll-Like/genética , Receptor 7 Toll-Like/metabolismo , Receptores Toll-Like/antagonistas & inibidores , Receptores Toll-Like/metabolismo
19.
Mol Med Rep ; 7(4): 1143-8, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23404565

RESUMO

In the present study, the effects of chordin­like protein 1 (CHRDL1) overexpression, together with bone morphogenetic protein­4 (BMP­4) treatment, on the differentiation of rat spinal cord­derived neural stem cells (NSCs) was investigated. Adult rat spinal cord­derived NSCs were cultured in serum­free medium. The recombined eukaryotic expression vector pSecTag2/Hygro B­CHRDL1 was transfected into adult rat spinal cord­derived NSCs using a lipid­based transfection reagent and protein expression was assessed by western blot analysis. Differentiation of transfected NSCs following BMP­4 treatment was determined by immunocytochemistry. The percentage of microtubule­associated protein­2 (MAP­2)­positive cells in the BMP­4­treated (B) group was found to be significantly lower compared with that in the non­transfected control (N) group. The percentage of MAP­2­positive cells in the pSecTag2/Hygro B­CHRDL1­transfected, BMP­4­treated group was identified to be significantly higher compared with that in group B, however, no significant difference was observed between group N and the transfected, non­BMP­4­treated control group. The current study indicates that CHRDL1 protein antagonizes BMP­4 activity and induces spinal cord­derived NSCs to differentiate into neurons.


Assuntos
Proteína Morfogenética Óssea 4/genética , Proteínas Morfogenéticas Ósseas/metabolismo , Glicoproteínas/genética , Peptídeos e Proteínas de Sinalização Intercelular/genética , Proteínas do Tecido Nervoso/genética , Células-Tronco Neurais/citologia , Neurônios/metabolismo , Animais , Proteína Morfogenética Óssea 4/administração & dosagem , Proteína Morfogenética Óssea 4/metabolismo , Proteínas Morfogenéticas Ósseas/administração & dosagem , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Proteínas do Olho/biossíntese , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Glicoproteínas/biossíntese , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/biossíntese , Proteínas do Tecido Nervoso/biossíntese , Células-Tronco Neurais/metabolismo , Neurônios/citologia , Ratos , Medula Espinal/citologia , Medula Espinal/metabolismo
20.
PLoS One ; 7(12): e51040, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-23251420

RESUMO

BACKGROUND: Vitiligo is characterized by the death of melanocytes in the skin. This is associated with the presence of T cell infiltrates in the lesional borders. However, at present, there is no detailed and systematic characterization on whether additional cellular or molecular changes are present inside vitiligo lesions. Further, it is unknown if the normal appearing non-lesional skin of vitiligo patients is in fact normal. The purpose of this study is to systematically characterize the molecular and cellular characteristics of the lesional and non-lesional skin of vitiligo patients. METHODS AND MATERIALS: Paired lesional and non-lesional skin biopsies from twenty-three vitiligo patients and normal skin biopsies from sixteen healthy volunteers were obtained with informed consent. The following aspects were analyzed: (1) transcriptome changes present in vitiligo skin using DNA microarrays and qRT-PCR; (2) abnormal cellular infiltrates in vitiligo skin explant cultures using flow cytometry; and (3) distribution of the abnormal cellular infiltrates in vitiligo skin using immunofluorescence microscopy. RESULTS: Compared with normal skin, vitiligo lesional skin contained 17 genes (mostly melanocyte-specific genes) whose expression was decreased or absent. In contrast, the relative expression of 13 genes was up-regulated. The up-regulated genes point to aberrant activity of the innate immune system, especially natural killer cells in vitiligo. Strikingly, the markers of heightened innate immune responses were also found to be up-regulated in the non-lesional skin of vitiligo patients. CONCLUSIONS AND CLINICAL IMPLICATIONS: As the first systematic transcriptome characterization of the skin in vitiligo patients, this study revealed previously unknown molecular markers that strongly suggest aberrant innate immune activation in the microenvironment of vitiligo skin. Since these changes involve both lesional and non-lesional skin, our results suggest that therapies targeting the entire skin surface may improve treatment outcomes. Finally, this study revealed novel mediators that may facilitate future development of vitiligo therapies.


Assuntos
Imunidade Inata/genética , Melanócitos/imunologia , Pele/imunologia , Vitiligo/genética , Adolescente , Adulto , Idoso , Feminino , Expressão Gênica/imunologia , Perfilação da Expressão Gênica , Humanos , Queratinócitos/imunologia , Queratinócitos/metabolismo , Queratinócitos/patologia , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Células Matadoras Naturais/patologia , Masculino , Melanócitos/metabolismo , Melanócitos/patologia , Pessoa de Meia-Idade , Pele/metabolismo , Pele/patologia , Linfócitos T/imunologia , Linfócitos T/metabolismo , Linfócitos T/patologia , Vitiligo/imunologia , Vitiligo/patologia
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