Data/hora: 02/06/2024 14:53:20

Provedor de dados: | 56
País: | Brazil
Título: | Lentiviral-mediated RNAi targeting caspase-3 inhibits apoptosis induced by serum deprivation in rat endplate chondrocytes in vitro
Autores: | Ding,L.; Wu,J.P.; Xu,G.; Zhu,B.; Zeng,Q.M.; Li,D.F.; Lu,W.
Data: | 2014-06-01
Ano: | 2014
Palavras-chave: | Cartilaginous endplate; Chondrocytes; RNA interference; Apoptosis; Caspase-3; Serum deprivation.
Resumo: | Current studies find that degenerated cartilage endplates (CEP) of vertebrae, with fewer diffusion areas, decrease nutrient supply and accelerate intervertebral disc degeneration. Many more apoptotic cells have been identified in degenerated than in normal endplates, and may be responsible for the degenerated grade. Previous findings suggest that inhibition of apoptosis is one possible approach to improve disc regeneration. It is postulated that inhibition of CEP cell apoptosis may be responsible for the regeneration of endplates. Caspase-3, involved in the execution phase of apoptosis, is a candidate for regulating the apoptotic process. In the present study, CEP cells were incubated in 1% fetal bovine serum. Activated caspases were detected to identify the apoptotic pathway, and apoptosis was quantified by flow cytometry. Lentiviral caspase-3 short hairpin RNA (shRNA) was employed to study its protective effects against serum deprivation. Silencing of caspase-3 expression was quantified by reverse transcription-polymerase chain reaction and Western blots, and inhibition of apoptosis was quantified by flow cytometry. Serum deprivation increased apoptosis of rat CEP cells through activation of a caspase cascade. Lentiviral caspase-3 shRNA was successfully transduced into CEP cells, and specifically silenced endogenous caspase-3 expression. Surviving cells were protected by the downregulation of caspase-3 expression and activation. Thus, lentiviral caspase-3 shRNA-mediated RNAi successfully silenced endogenous caspase-3 expression, preventing inappropriate or premature apoptosis.
Tipo: | Info:eu-repo/semantics/article
Idioma: | Inglês
Identificador: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2014000600445
Editor: | Associação Brasileira de Divulgação Científica
Relação: | 10.1590/1414-431X20143198
Formato: | text/html
Fonte: | Brazilian Journal of Medical and Biological Research  v.47 n.6 2014
Fonte: | info:eu-repo/semantics/openAccess