[In vivo transfection of cis element 'decoy' against nuclear factor-kappaB binding site modulates function of T lymphocytes in asthma].
2004
Objective To investigate the effect of nuclear factor-κB(NF-κB) decoy oligodeoxynucleotide(ODN) on proliferation, apoptosis and cytokine synthesis of T lymphocytes from asthmatic patients Methods T lymphocytes were divided into four groups,a normal control group (A group), an asthma control group (B group), a NF-κB cis decoy ODN group (B 1 group) and a scrambled ODN group (B 2 group) B 1 and B 2 groups were transfected by cationic lipofectamine to the latter two groups respectively The proliferation of T lymphocytes was measured by MTT and the apoptosis of them was measured by flow cytometry The expression levels of interleukin 5(IL-5) mRNA and protein were detected with cell hybridization in situ and enzyme-linked immunosorbent assay(ELISA) The expression levels of inducible nitric oxide synthase(iNOS) were measured by reverse transcription-poly merase chain reaction(RT-PCR) and Western blot Results The difference of proliferation rate between B 1 group (0 220±0 020) and B group(0 340±0 030) was significant ( P 0 05) The difference of apoptosis rate between B 1 group(10 8±1 3) and B group(8 1±1 2) was also significant( P 0 05) The expression levels of IL-5 mRNA and protein in B 1 group(21±4,24±4)were significantly different from those in B group (33±4,54±10, P 0 05) The differences of iNOS mRNA and protein levels between B 1 group (0 33±0 05, 782±117)and B group (0 75±0 13,1185±230) were significant ( P 0 05) However, these indices in B 2 group showed no difference to those in B group( P 0 05) Conclusions NF-κB decoy ODN can reduce the abnormally increased proliferation of T lymphocytes in asthma and increase the abnormally decreased apoptosis of these cells, while decrease the abnormally increased levels of cytokine and enzyme in asthmatic T lymphocytes These may be the mechanisms underlying its potential therapeutic effects in asthma
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