[Regulation of NF-κB signal transduction pathway on cytokines in cultured nasal epithelium].
OBJECTIVE
To explore the activation and regulation of nuclear factor-kappa B (NF-κB) on transcription of cytokines in cultured lipopolysaccharide (LPS)-induced nasal epithelial cells.


METHODS
Normal sphenoid mucosa epithelium from 11 patients who accepted pituitary tumor surgery via trans-sphenoid approach was separated and cultured without serum. The epithelium of the third or the forth passage was induced with LPS. Wedelolactone, a blocking agent of NF-κB was used at the same time. An electrophoretic mobility shift assay was used to detect DNA-binding activity of NF-κB. The reverse transcription-polymerase chain reaction was used to evaluate mRNA of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), IL-5, IL-6, IL-8, granulocyte-macrophage colony stimulating factor (GM-CSF), regulated on activation, normal T expressed and secreted (RANTES), eotaxin, eotaxin-2, vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Statistical analysis was performed using SPSS17.0 software.


RESULTS
DNA binding activity of NF-κB and mRNA of IL-1β, IL-8 and COX-2 increased in cultured LPS-induced nasal epithelial cells (relative values were 1.013 ± 0.144, 0, 0, 0 respectively in control group and relative ones of LPS-induced were 2.050 ± 0.305, 1.057 ± 0.041, 0.950 ± 0.042, 0.117 ± 0.012 respectively). There was significant difference between the control group and LPS-induced nasal epithermal cells group (P values were 0.004, 0.000, 0.000, 0.000 respectively). Corresponding expression of NF-κB, IL-1β, IL-8 and COX-2 decreased after the addition of Wedelolactone (relative values were 0.917 ± 0.188, 0.180 ± 0.008, 0, 0 respectively). There was significant difference between the LPS-induced nasal epithelial cells group and the Wedelolactone-addition group (P values were 0.002, 0.000, 0.000, 0.000 respectively). But the expression of mRNA of other factors were 0 in all groups.


CONCLUSIONS
The NF-κB signal transduction pathway was involved in the transcriptional regulation of IL-1β, IL-8 and COX-2 in cultured LPS-induced nasal epithelial cells.
Department of Otorhinolaryngology Head and Neck Surgery, Third Affiliated Hospital of SUN Yat-sen University, Guangzhou 510630, China.
Ji-hong
JH
Yang
Yuan
Y
Li
Ge-hua
GH
Zhang
Qin-tai
QT
Yang
D002478:Cells, Cultured; D051546:Cyclooxygenase 2 / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D053583:Interleukin-1beta / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism*; D009297:Nasal Mucosa / Q000166:cytology / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D015398:Signal Transduction
D016207:Cytokines; D053583:Interleukin-1beta; D016209:Interleukin-8; D016328:NF-kappa B; D012333:RNA, Messenger; D051546:Cyclooxygenase 2