1.The experimental study on effects of nuclear factor-κBp65 antisense oligonueleotide on liver fibrosis
Yu ZHOU ; Kequan CHEN ; Shicai YE ; Zhuang WANG ; Jian LIANG ; Ronghuo LIU ; Zhiling TANG ; Hao WANG ; Xiao FENG ; Wengtao YE
Chinese Journal of Digestion 2009;29(4):254-257
Objective To investigate the effect of the nuclear factor (NF)-κBp65 antisense oligonucleotide (ASODN) on NF-κB activity and expression of interleukin(IL)-6 in hepatic stellate cells (HSC). Methods The HSC were separated from rats and cultured. The toxicity of NF-κBp65 ASODN on HSC were detected by Trypan blue exclusion staining and the NF-κB activity was determined by EMSA. The expressions of IL-6 mRNA and protein were meaured by RT-PCR and ELISA, respectively. Results In vitro, no toxicity of ASODN on HSC was observed at the concentrations of 0. 001 to 1.0 μmol/L. NF-κB activity was increased after stimulating HSC with tumor necrosis factor (TNF)α, whereas it was weakened in a dose dependent manner when HSC were cultured with ASODN (concentration from 0. 001 to 1.0 μmol/L). At the same time, the expressions of IL-6 mRNA and protein induced by TNFα were decreased after transfected with ASODN at concentrations of 0.001- 1. 0 μmol/L in a dose dependent manner. Conclusion ASODN may specifically inhibit either the activiy of NF-κB or expression of IL-6, which provides the theoretical basis that ASODN may use to treat fibrosis of the liver.
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