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1.
Lett Appl Microbiol ; 70(2): 118-127, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31782817

RESUMO

The aim of this study was to determine whether Lactobacillus rhamnosus GG (LGG) components (surface layer protein, SLP; genomic DNA, gDNA; unmethylated cytosine-phosphate-guanine-containing oligodeoxynucleotide, CpG-ODN), alone or in combination, could affect immunomodulation, and evaluate the signalling mechanism in mouse macrophage RAW264.7 cells challenged with lipopolysaccharide (LPS). LGG components were used to treat cells before LPS stimulation. Cytokine and Toll-like receptor (TLR) expression were assessed using real-time quantitative PCR (RT-qPCR). Mitogen-activated protein kinase (MAPK), extracellular regulated protein kinase (ERK) and nuclear factor-kappa B (NF-κB) signalling pathways were evaluated using immunoblots and immunofluorescence. SLP or SLP + gDNA pre-treatment significantly reduced the LPS-induced mRNA expression of tumour necrosis factor alpha (TNF-α). Pre-treatment with LGG single components (SLP, gDNA or CpG) or their combinations (SLP + gDNA or SLP + CpG) significantly decreased the LPS-induced interleukin-6 (IL-6) mRNA level (P < 0·05). Pre-treatment with SLP or gDNA, alone or in combination, significantly suppressed LPS-induced TLR2 and TLR4 mRNA levels (P < 0·05). SLP pre-treatment also significantly decreased the LPS-induced expression of TLR9 (P < 0·05). Pre-treatment with LGG single components or combinations significantly suppressed the LPS-induced phosphorylation levels of ERK (P > 0·05). In conclusion, pre-incubation with LGG components, singly or in combination, generally inhibited the activation of TLR, MAPK and NF-κB signalling pathways in LPS-stimulated cells, leading to attenuated inflammatory cytokine TNF-α and IL-6 production. These results indicate that nonviable probiotic LGG components exert an anti-inflammation effect on epithelial cells. SIGNIFICANCE AND IMPACT OF THE STUDY: Lactobacillus rhamnosus GG (LGG) is widely used as probiotics. However, its main components are not well known for affecting immunomodulation. This study investigated the effects of pre-treatments with different components such as surface layer protein, genomic DNA and unmethylated cytosine-phosphate-guanine-containing oligodeoxynucleotides, alone or in combination on immunomodulation, and evaluated the signalling mechanism in mouse macrophage RAW264.7 cells challenged with lipopolysaccharide. Pre-incubation with components alone or in combination generally inhibited the activation of Toll-like receptor, mitogen-activated protein kinases, extracellular regulated protein kinases and nuclear factor-kappa B signalling pathways in lipopolysaccharide-stimulated cells, which generally leads to attenuated inflammatory cytokine interleukin-6 and tumour necrosis factor alpha production. These results indicate that nonviable probiotic LGG components exert an anti-inflammation effect on epithelial cells.


Assuntos
Lacticaseibacillus rhamnosus/imunologia , Lacticaseibacillus rhamnosus/metabolismo , Lipopolissacarídeos/toxicidade , Sistema de Sinalização das MAP Quinases/fisiologia , Macrófagos/imunologia , Animais , Composição de Bases/genética , Linhagem Celular , Citocinas/metabolismo , DNA Bacteriano/genética , Células Epiteliais , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Interleucina-6/metabolismo , Lacticaseibacillus rhamnosus/genética , Camundongos , NF-kappa B/metabolismo , Oligonucleotídeos/genética , Probióticos/farmacologia , Células RAW 264.7 , RNA Mensageiro/metabolismo , Receptores Toll-Like/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
2.
Cell Prolif ; 46(4): 447-56, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23829318

RESUMO

OBJECTIVES: Mesenchymal stem cells (MSCs) are a reliable resource for tissue regeneration, but their molecular mechanisms of differentiation and proliferation remain unclear; this situation has restricted use of MSCs to a limited number of applications. A previous study of ours found a member of the epidermal growth factor family, epiregulin (EREG), to be involved in regulation of MSC differentiation. In the present study, we have used human dental stem cells from the apical papilla (SCAPs) to investigate the role of EREG on proliferation of MSCs. MATERIALS AND METHODS: SCAPs were isolated from apical papillae of immature third molars. Retroviral short hairpin RNA (shRNA) was used to silence EREG gene expression, and human recombinant EREG protein was used to stimulate SCAPs. SCAP proliferation was examined using tetrazolium dye colorimetric assay/cell growth curve. Western blotting was performed to detect expressions of extracellular signal-regulated protein kinases 1 and 2 (Erk1/2), mitogen-activated protein kinases 1 and 2 (MEK1/2), protein kinase B (Akt), p38 mitogen-activated protein kinase (p38 MAPK) and c-Jun N-terminal kinase (JNK). RESULTS: Depletion of EREG with shRNA inhibited SCAP proliferation and repressed phosphorylation of Erk1/2 and JNK. Human recombinant EREG protein promoted cell proliferation and enhanced Erk1/2, MEK and JNK phosphorylation in SCAPs. Furthermore, blocking MEK/Erk signalling with specific Erk1/2 inhibitor PD98059, or JNK signalling with specific inhibitor SP600125, abolished effects of EREG on cell proliferation. CONCLUSION: These findings indicate that EREG could enhance cell proliferation in dental tissue-derived MSCs by activating MEK/Erk and JNK signalling pathways.


Assuntos
Fator de Crescimento Epidérmico/metabolismo , Fator de Crescimento Epidérmico/farmacologia , Sistema de Sinalização das MAP Quinases , Células-Tronco/metabolismo , Adolescente , Diferenciação Celular/fisiologia , Processos de Crescimento Celular/efeitos dos fármacos , Processos de Crescimento Celular/fisiologia , Células Cultivadas , Papila Dentária/citologia , Papila Dentária/efeitos dos fármacos , Papila Dentária/enzimologia , Papila Dentária/fisiologia , Fator de Crescimento Epidérmico/deficiência , Fator de Crescimento Epidérmico/genética , Epirregulina , Humanos , Fosforilação , Proteínas Proto-Oncogênicas c-akt/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Interferente Pequeno/administração & dosagem , RNA Interferente Pequeno/genética , Proteínas Recombinantes/farmacologia , Células-Tronco/citologia , Células-Tronco/efeitos dos fármacos , Células-Tronco/fisiologia , Adulto Jovem , Proteínas Quinases p38 Ativadas por Mitógeno/genética , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
3.
J Phys Chem B ; 110(12): 5966-9, 2006 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-16553404

RESUMO

High-aspect-ratio and single-crystal aluminum borate (Al(18)B(4)O(33)) nanowire bundles with an ordered orientation were synthesized by using an innovative sucrose-assisted growth process. The process involves the dehydration and polycondensation of aluminum borate-sucrose solution to form a highly viscous precursor. The sucrose plays a crucial role in the growth of the nanowire bundles by supporting as a polymeric substrate and a type of adhesive template. Electron microscopy was used to characterize the high-aspect-ratio nanowire bundles. A possible growth mechanism for the nanowire bundles is proposed.

4.
J Nanosci Nanotechnol ; 5(6): 932-6, 2005 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-16060156

RESUMO

A methanol-thermal method has been developed to fabricate one-dimensional composite nanowires by coating multiwalled carbon nanotubes with lanthanum oxide and iron oxide, respectively. The coating structure and composition have been investigated by transmission electron microscopy and the X-ray energy dispersive spectrum. Magnetic measurement for the iron oxide coatings indicated that the coercivity has been enhanced after coating. The method reported here provides a novel procedure for the fabrication of one-dimensional composite nanostructures.


Assuntos
Cristalização/métodos , Compostos Férricos/química , Lantânio/química , Magnetismo , Nanotubos de Carbono/química , Nanotubos de Carbono/ultraestrutura , Óxidos/química , Instalação Elétrica , Compostos Férricos/análise , Temperatura Alta , Lantânio/análise , Metanol/química , Conformação Molecular , Nanotubos de Carbono/análise , Óxidos/análise
5.
J Phys Chem B ; 109(27): 13060-2, 2005 Jul 14.
Artigo em Inglês | MEDLINE | ID: mdl-16852623

RESUMO

A novel route was proposed to completely coat aluminum borate nanowires by in situ providing the precursor for BN coating. Uniformly BN-coated Al18B4O33 nanowires could be obtained by the reaction of Al4B2O9 nanowires with ammonia at high temperature. The high-temperature unstable Al4B2O9 nanowires were converted into Al18B4O33 nanowires, simultaneously evaporated boron oxide. The reaction between the in situ generated vapors and ammonia ensures that the BN layers are attached tightly on the surface of the as-formed Al18B4O33 nanowires.

6.
J Nanosci Nanotechnol ; 4(8): 1071-5, 2004 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-15656205

RESUMO

Comparative study on the diameter distribution of MgO nanowires has been carried out. MgO nanowires could be synthesized by the direct reaction between metallic magnesium and silica, and the obtained nanowires have diameters ranging from 50 to 200 nm and lengths of several hundreds nanometers, exhibiting a straight wire. The diameter can be downscaled to smaller than 50 nm, and the nanowire exhibits a curved and twisted one-dimensional structure with lengths up to several micrometers, when a fine support catalyst was used as the reactant. The diameter-controlled growth mechanism was also explained in this work.


Assuntos
Cristalização/métodos , Óxido de Magnésio/química , Magnésio/química , Nanotecnologia/métodos , Nanotubos/química , Nanotubos/ultraestrutura , Dióxido de Silício/química , Teste de Materiais , Conformação Molecular , Nanotubos/análise , Tamanho da Partícula , Propriedades de Superfície
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