The Effects Of Perfluorocarbon On LPS-induced Type Ⅱ Alveolar Epithelial Cells And Suppression Of MAPK Pathways

Objective：Acute lung injury/Acute respiratory distress syndrome (ALI/ARDS) is acommon and critical pulmonary complication with high incidence and mortality rate.ALI is characterized by excessive neutrophil infiltration, release of inflammatorymediators and destruction of the alveolar–capillary membrane causing severe impact onpulmonary gas exchange. Perfluorocarbon has been investigated as liquid breathingmedia as part of the treatment for ALI or ARDS since it can improve oxygenation,respiratory function and block neutrophil infiltration. This study was designed toinvestigate the effect of perfluorocarbons(PFCs) on MIP-1α， MIP-2, andMCP-1mRNA expression induced by LPS-stimulated A549cells and its relationshipwith mitogen-activated protein kinases (MAPK)pathways.Methods:(1) We use PFC to modulate LPS-induced A459cells, which are dividedrandomly into4groups:(1) Control group;(2) LPS group;(3) LPS+PFC group; and (4)PFC group. The mRNA and protein expression level of MIP-1α、MIP-2、MCP-1ineach group at1,2,4,6,8and12hr, were detected by real time PCR and western blot,respectively.(2) The cell samples are harvested at1,2,4,6,8and12hr for Western blotdetection of phosphorylation of ERK1/2, p38MAPK, JNK, ATF-2, c-jun proteins andtotal of ERK1/2, p38MAPK and JNK proteins.(3) Preincubation of cells with p38MAPK inhibitor SB203580, MEK1/2inhibitor U0126, and SAPK/JNKSP600125single or combination, MIP-2mRNA are assayed by RT-qPCR at1,6, and12hr.Results:(1) PFC attenuates mRNA and protein expression of MIP-2in LPS-inducedA549cells;(2) PFC effectively suppresses MAPK activation by inhibiting thephosphorylation of p38MAPK， ERK1/2, and SAPK/JNK. PFC also attenuates thephosphorylation of ATF-2and c-jun.(3) Preincubation of cells with U0126, SP600125, and SB203580single or combination has significantly downregulated MIP-2expression in LPS stimulated A549cells.(4) PFC attenuates LPS-induced production ofMIP-2by inhibiting the phosphorylation of MAPK, ATF-2and c-jun. It might be thepotential molecular mechanism for therapeutic inflammation disease by PFC.