Inhibition of tumor necrosis factor-α expression in macrophages with vector-mediated RNA interference / 中华消化杂志

ABSTRACT

Objective To investigate the inhibitory effect of small interference RNA(siRNA)on the exDression of tumor necrosis factor(TNF)-oligo nucleotide fragments were designed and synthesized according to the sequence of TNF-insert downstream to H1 promoter to construct plasmids pHS-A and pHS-B for expression of short hairpin RNA(shRNA).Then the recombinant pSilence3.1-TNF-phages(RAW264.7)by lipofectamin 2000.The inhibition of TNF-examined bv real-time PCR and ELISA,respectively.Results After LPS stimulating 6 hours,TNF-expression was increased in a specific time-dependent manner,and reached high peak at 9-12 hours.The macrophages treated with plasmids pHS-A showed a significant decrease in TNF- mRNA(0.021 34±0.009 60)and protein(149.93,P＜0.01)compared with untransfected group[TNF-mRNA0.021 34±0.009 60,protein1 922.30±149.05]pg/ml].The relative rate of inhibition was 83.3％.No inhibitory effects was found in plasmids pHS-B and control group.Conclusions LPS stimulation results in a increasing expression of TNF-of TNF-