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1.
Artigo em Chinês | MEDLINE | ID: mdl-27220436

RESUMO

OBJECTIVE: To explore the effect of paraquat (PQ) on autophagy in human embryonic neural progenitor cells. METHODS: Using ReNcell CX cell model. After treatment with various concentration (0.00, 1.00, 10.00 and 100.00 µmol/L) of PQ, CCK8 assay was used to detect the cell viability, the transmission electron microscope was used to observe the the cell ultrastructure, the real-time polymerase chain reaction (RT-PCR) was adopted to detect mRNA expression of autophagy related genes which including LC3, Atg12, Atg5, beclin1, Atg7 and mTOR and apoptosis related genes Bax and Bcl-2. RESULTS: The cell viability was significantly inhibited after administered with 100.00 µmol/L of PQ (P<0.01). The mRNA expression of Beclin1 was significantly up-regulated and the emergency of autophagosome were observed at the concentration of 1.00 µmol/L group, while mild cell apoptosis, significantly up-regulated Atg5, Atg8, Atg7 and Atg12 mRNA expression as well as down-regulated expression of mTOR and Bax were detected at the 10.00 µmol/L of PQ group, howere, the obvious apoptosis and the up-regulated mRNA expression of mTOR and Bax were observed at the 100.00 µmol/L of PQ group, the mRNA expression of Bcl-2 were all down-regulated after administered with 1.00, 10.00 and 100.00 µmol/L of PQ and reached the lowest level at the concentration of 10.00 µmol/L. CONCLUSION: PQ can induced autophagy at the low concentration in ReNcell CX cell and autophagy might serve as a protective mechanism to ameliorate PQ-induced cytotoxic effects but apoptosis will be induced at the 100 µmol/L concentration.


Assuntos
Autofagia , Células-Tronco Embrionárias Humanas , Células-Tronco Neurais , Apoptose , Proteínas Reguladoras de Apoptose , Sobrevivência Celular , Regulação para Baixo , Humanos , Paraquat , Serina-Treonina Quinases TOR , Regulação para Cima
2.
Clin Exp Dermatol ; 40(8): 916-9, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26224166

RESUMO

Hidradenitis suppurativa (HS) is a chronic disease of follicular occlusion. It involves the axilla, groin, perianal and perineal regions, and is characterized by recurrent draining sinuses, skin abscesses and disfiguring scars. Loss-of-function mutations in the genes encoding γ-secretase have been identified as a cause of HS. We collected skin samples from three patients with HS from a Chinese family carrying a NCSTN mutation (c.1258C>T (p.Q420X)) and three unrelated healthy controls (HCs). Expression level of nicastrin in skin tissue and cultured keratinocytes and fibroblasts of patients and HCs was determined by real-time quantitative PCR and western blotting. We found that the mRNA and protein levels of nicastrin were significantly reduced in the whole skin, epidermis, dermis, and cultured keratinocytes and fibroblasts compared with HCs. Therefore, we conclude that haploinsufficiency of the NCSTN gene caused by the nonsense mutation c.1258C>T (p.Q420X) contributes to the occurrence of HS in this family.


Assuntos
Secretases da Proteína Precursora do Amiloide/genética , Códon sem Sentido , Haploinsuficiência/genética , Hidradenite Supurativa/genética , Glicoproteínas de Membrana/genética , Povo Asiático/genética , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
3.
Eur Cytokine Netw ; 26(1): 10-4, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25990837

RESUMO

Endothelin-1 (ET-1) acts as a key regulator of vasoconstriction and fibrosis. Many previous studies have focused on the role of ET-1 in scleroderma (systemic sclerosis, SSc). We investigated the effects of ET-1 on the production of extracellular matrix in SSc and normal skin fibroblasts. Primary cultured dermal fibroblasts from SSc patients and healthy controls were treated with ET-1 (25 ng/mL) for 0 min, 15 min, 1 h, 24 h, 48 h and 72 h, respectively. Our results showed that, in SSc fibroblasts, ET-1 upregulated collagen type I, connective tissue growth factor (CTGF), type I plasminogen activator inhibitor (PAI-1) and pAkt in a time-dependent manner within 72 h; in normal fibroblasts, 25 ng/mL ET-1 stimulation correlated with high levels of CTGF, PAI-1 and pAkt. The secretion of fibronectin (FN), collagen type I, and PAI-1 is markedly increased in the supernatant of both SSc fibroblasts and normal fibroblasts. Furthermore, ET-1 phosphorylates Smad2 and Smad3 in normal fibroblasts, but not in SSc fibroblasts. In conclusion, our results demonstrated that ET-1 may induce fibrosis in dermal fibroblasts through Akt signals.


Assuntos
Endotelina-1/farmacologia , Fibroblastos/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/genética , Escleroderma Sistêmico/genética , Pele/efeitos dos fármacos , Adulto , Estudos de Casos e Controles , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Fator de Crescimento do Tecido Conjuntivo/genética , Fator de Crescimento do Tecido Conjuntivo/metabolismo , Endotelina-1/metabolismo , Feminino , Fibroblastos/metabolismo , Fibroblastos/patologia , Fibronectinas/biossíntese , Fibronectinas/metabolismo , Fibrose , Regulação da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Fosforilação , Inibidor 1 de Ativador de Plasminogênio/genética , Inibidor 1 de Ativador de Plasminogênio/metabolismo , Cultura Primária de Células , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologia , Escleroderma Sistêmico/metabolismo , Escleroderma Sistêmico/patologia , Transdução de Sinais , Pele/metabolismo , Pele/patologia , Proteína Smad2/genética , Proteína Smad2/metabolismo , Proteína Smad3/genética , Proteína Smad3/metabolismo
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