Kinetics of nuclear factor ?B in LPS stimulated rat alveolar macrophages and its regulative effect on tumor necrosis factor ? expression
- VernacularTitle:核因子-?B在内毒素刺激大鼠肺泡巨噬细胞中的动态变化及其在肿瘤坏死因子-?表达中的作用
- Author:
Chunxue LI
;
Jianxin JIANG
;
Youan SHAN
- Publication Type:Journal Article
- Keywords:
Nuclear factor-kappa B;
Lipopolysaccharides;
Tumor necrosis factor-alpha;
Macrophage, alveolar;
Decoy-phosphorothioate oligodeoxynucleotides
- From:
Chinese Journal of Trauma
1993;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the activation of nuclear factor ?B (NF-?B) induced by lipopolysaccharides (LPS) in rat alveolar macrophages (AM) and its regulative role in tumor necrosis factor (TNF-?) expression. Methods The dynamic activity changes of NF-?B DNA induced by LPS (E.coli 026:B6) were determined with electrophoretic mobility shift assay (EMSA). The phosphorothioate oligodeoxynucleotides (S-ODN) decoy was transfected into AM 12 hours prior to LPS stimulation. The effect of NF-?B S-ODN decoy on expression of TNF-? in AM stimulated by LPS were measured with enzyme-linked immunosorbent assay (ELISA) kit. Results NF-?B could be activated remarkably after 0.5 hour of LPS stimulation at concentration of 100 ng/ml, reached the highest level 1 hour after LPS stimulation and gradually decreased. But the activation of NF-?B could last at least 8 hours. The dose for LPS stimulation was related to activation of NF-?B in a dose-dependent fashion, ie, the activation of NF-?B gradually strengthened with dose increase of LPS. Supershift assays proved that p50 and p65 were involved in the activation of NF-?B. NF-?B S-ODN decoy could markedly (not completely) inhibit LPS-induced TNF-? production. Conclusions NF-?B plays an important role in LPS induced inflammatory response. However, entire inhibition of the activity of NF-?B can not completely prevent TNF-? expression induced by LPS in rat AM, which implies that other nuclear factors may participate in TNF-? expression.
