Morphology,function And Expression Of P311 Change In Lung Cells In Hyperoxia Environment

Objective: The changes of the morphology,apoptosis level and intracellular reactive oxygen species(ROS)in lung cells were observed under different times in air and hyperoxia environment.And the changes of P311 m RNA and protein in type ΙΙ alveolar epithelial cells(AECⅡ)were determined to explore morphological and functional changes of lung cells and the role of P311 in the mechanism of high oxygen lung injury.Methods: AECⅡ and lung fibroblasts(LFs)from fetal mice were isolated for primary cell culture.AECⅡ was identified by transmission electron mircroscopy and immunofluoresence method.LFs was identified by immunohistochemistry method.And the cell model of hyperoxia induced lung injury was constructed.Two kinds of cells were divided into air group(oxygen volume fraction,0.21)and hyperoxia groups(oxygen volume fraction,0.9).Inverted microscope was used to observe the morphology and migration rate in different groups of AECⅡ and LFs cells.Flow cytometry was conducted to detect the intracellular ROS levels and apoptosis in the experimental and control groups of two kinds of cells at different times.Immunofluorescence method was used to detect the changes of a-SMA fluorescence intensify in two groups of LFs.Real-time PCR and Western Blot were used to detect the expression levels of P311 m RNA and P311 protein at different times in AECⅡ.Results: With the hyperoxia time prolonged,the migration level of AECⅡ in hyperoxia group decreased(P<0.01).Intracellular ROS and apoptosis level and the expression of P311 protein and m RNA was significantly increased(P<0.01).Besides,the morphological of LFs were changed obviously and the migration level increased in the hyperoxia group(P<0.01).The content of Intracellular ROS increased significantly(P<0.01).The fluorescence intensify of a-smooth muscle actin(a-SMA)in the hyperoxia group of LFs was obviously increased,which suggests the transformation from LFs to myofibroblasts(MFs).Conclusion: High concentration of oxygen causes lung cells to grow poorly.The migration ability of AEC Ⅱdrop,but LFs increases.The mass production of intracellar ROS eventually leads to apoptosis of two kinds of cells.P311 expression increased suggests that P311 may participate in the repair of hyperoxia induced lung injury.The increase of fluorescence intensify of a-SMA indicate the transformation from LFs to myofibroblasts.