Effects Of Perlfuorocarbons On Pulmonary Surfactant Protein In Llpopolysaccharide-induced A549Cells

Objective: To investigate the effects of perfluorocarbons(PFC) on pulmonarysurfactant proteins in lipopolysaccharide(LPS)-induced A549cells.Materials and Methods: The experiments was divided into four groups:(1)Control group(C): DMEM containing A549cells at high density (106cells/ml);(2)LPS group (L): DMEM mixed with LPS at a concentration of10ug/ml containingA549cells at a density of106cells/ml;(3) PFC group (P): DMEM mixed with PFC ata ratio (v/v) of10%containing A549cells at a density of106cells/ml;(4) LPS+PFCgroup (L+P): DMEM mixed with LPS at a concentration of10ug/ml and PFC at a ratio(v/v) of10%containing A549cells at a density of106cells/ml. Each group wasobserved at6time points:1h,2h,3h,4h,5h,6h, respectively, and each point wasprepared of total volume of1ml mixture which was exposed to ultrasonic energy for5seconds for3times with an interval of20seconds. The cells were harvested followedby centrifugation with5000rpm×5min and stored at-20℃.Part1:(1) The mRNA expression levels in cell lysates of IL-1β,IL-6and TNF-α ofgroup C and group L were examined by Real-time Quantitative PCR (q-PCR)respectively;(2) The mRNA expression levels of SP-D and TTF-1of group C andgroup F(including PFC at a volume ratio (v/v) of10%,30%,50%,70%respectively)were examined by q-PCR.Part2:Surfactant proteins in cell lysates at each time point of all group wereexamined by q-PCR,Western Blot and ELISA respectively.Part3:The mRNA expression levels in cell lysates of ABCA3and TTF-1at eachtime point of all group were examined by q-PCR respectively.Results:(1) The mRNA expression levels of IL-1β,IL-6and TNF-α in group Lwere significantly higher than those in group C.(2) Compared with the group L,the expression levels of mRNA and surfactant proteins of SP-A at2h, SP-B,SP-C andSP-D at3h were increased in group L+P, respectively.(3) The mRNA expression levelsof ABCA3at2h and3h, TTF-1at3h in group L+P were significantly higher than thosein group L, respectively.Conclusion: PFC could increase the expression of surfactant proteins inLPS-induced ALI A549cells model, which suggested that PFC could therapy the acutelung injury by increasing surfactant proteins expression.