[Activation of nuclear factor kappaB signaling pathway in human osteoblasts responses to Staphylococcus aureus in vitro].
OBJECTIVE
To investigate whether S. aureus could activate NF-κB signaling pathway in human osteoblasts.


METHODS
Immunoblot and electrophoretic mobility shift assay were used to detect the degradation of I-κBα and activation of NF-κB in human osteoblasts following infection with S.aureus, respectively, and there were investigated the activated state of NF-κB signaling pathway in human osteoblasts. In addition, enzyme-linked immunosorbent assay was used to measure the secretion of IL-6 in culture supernatants, which was represented as one of important cytokines in osteomyelitis, and an inhibitor of NF-κB, SN50, which was added to human osteoblasts culture prior to 1 hour at 50 µmol/L before the infection of S.aureus, was used to determine whether S.aureus-activated NF-κB signaling pathway regulates IL-6 secretion of human osteoblasts.


RESULTS
S.aureus could induce the degradation of I-κBα (I-κBα(15 min)/I-κBα(0 min) = 0.409 ± 0.245 and I-κBα(30 min)/I-κBα(0 min) = 0.061 ± 0.010) and activation of NF-κB in human osteoblasts in a time and dose-dependent manner following infection. In addition, the secretion of IL-6 in the supernatants of human osteoblasts ((2.17 ± 0.11) µg/L) was suppressed by 50 µmol/L SN50 compared to without the addition of SN50 ((3.58 ± 0.31) µg/L) (F = 174.25, P < 0.05).


CONCLUSIONS
S.aureus could activate NF-κB signaling pathway in human osteoblasts, which could regulate cytokines secretions of human osteoblasts.
Department of Orthopaedics, the Third Affiliated Hospital of Anhui Medical University, Hefer 230031, China.
Ren-de
RD
Ning
Xian-long
XL
Zhang
Li
L
Gong
Ye-jin
YJ
Zhou
Xiao-kui
XK
Guo
Qing-tian
QT
Li
D002478:Cells, Cultured; D006801:Humans; D015850:Interleukin-6 / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism*; D010006:Osteoblasts / Q000378:metabolism*; D015398:Signal Transduction; D013203:Staphylococcal Infections / Q000378:metabolism*
C508600:IL6 protein, human; D015850:Interleukin-6; D016328:NF-kappa B