摘要
Background Toll-like receptor-4 （TLR-4） is integrally involved in lipopolysaccharide （LPS） signaling and has a requisite role in the activation of nuclear factor-κB （NF-κB）. The exact mechanisms that lend perfluorocarbon （PFC） liquids a cytoprotective effect have yet to be elucidated. Therefore we examined in an in vitro model the cytoprotective effect of PFC on LPS-stimulated alveolar epithelial cellls （AECs）. Methods AECs （A549 cells, human lung adenocarcinoma cell line） were divided into four groups： control, PFC, LPS and LPS ＋ PFC （coculture group） groups. Intercellular adhesion molecule-1 （ICAM-1） was detected by ELISA, tumor necrosis factor-g （TNF-a） and interleukin-8 （IL-8） were detected by radioimmunological methods. The expression of TLR-4 mRNA and protein was detected by real time PCR and Western blotting, respectively. The activation of NF-κB was detected by Western blotting （proteins of I-κBa and NF-κB p65）. Results ICAM-1, TNF-a and IL-8 were significantly increased in LPS-stimulated AECs groups. The expression of TLR-4 mRNA and protein in LPS-stimulated groups was markedly increased. Meanwhile, NF-κB was activated as indicated by the significant degradation of IKB-a and the significant release of NF-κB P65 and its subsequent translocation into the nucleus. There were no significant effects of PFC alone on any of the factors studied while the coculture group showed significant downregulation of the secretion of ICAM-1, TNF-a and IL-8, the expression of TLR-4 rn^NA and the activity of NF-κB. Conclusions Taken together, our results demonstrate that LPS can induce AEC-related inflammatory injury via the activation of TLR-4 and subsequent activation of NF-κB. PFC is able to protect AECs from LPS-induced inflammatory injury by blocking the initiation of the LPS signaling pathway, which is indicated by the significant decrease of TLR-4 expression and NF-κB activation.
Background Toll-like receptor-4 （TLR-4） is integrally involved in lipopolysaccharide （LPS） signaling and has a requisite role in the activation of nuclear factor-κB （NF-κB）. The exact mechanisms that lend perfluorocarbon （PFC） liquids a cytoprotective effect have yet to be elucidated. Therefore we examined in an in vitro model the cytoprotective effect of PFC on LPS-stimulated alveolar epithelial cellls （AECs）. Methods AECs （A549 cells, human lung adenocarcinoma cell line） were divided into four groups： control, PFC, LPS and LPS ＋ PFC （coculture group） groups. Intercellular adhesion molecule-1 （ICAM-1） was detected by ELISA, tumor necrosis factor-g （TNF-a） and interleukin-8 （IL-8） were detected by radioimmunological methods. The expression of TLR-4 mRNA and protein was detected by real time PCR and Western blotting, respectively. The activation of NF-κB was detected by Western blotting （proteins of I-κBa and NF-κB p65）. Results ICAM-1, TNF-a and IL-8 were significantly increased in LPS-stimulated AECs groups. The expression of TLR-4 mRNA and protein in LPS-stimulated groups was markedly increased. Meanwhile, NF-κB was activated as indicated by the significant degradation of IKB-a and the significant release of NF-κB P65 and its subsequent translocation into the nucleus. There were no significant effects of PFC alone on any of the factors studied while the coculture group showed significant downregulation of the secretion of ICAM-1, TNF-a and IL-8, the expression of TLR-4 rn^NA and the activity of NF-κB. Conclusions Taken together, our results demonstrate that LPS can induce AEC-related inflammatory injury via the activation of TLR-4 and subsequent activation of NF-κB. PFC is able to protect AECs from LPS-induced inflammatory injury by blocking the initiation of the LPS signaling pathway, which is indicated by the significant decrease of TLR-4 expression and NF-κB activation.
