| Objective:The aim of the study was to investigate the expression of aquaporin-5in LPS induced AT-Ⅱ cells,exposed the effect of various Oxygen concentration, and its possible mechanism was also discussed.Methods:Primary cultured AT-Ⅱ cells were randomly divided into five groups:A blank control group,B LPS group,C Oxygen concentration of40%+LPS group,D Oxygen concentration of60%+LPS group, E Oxygen concentration of90%+LPS group. Towenty-Four hours later, supernatant fluid was used to assay the content of TNF-a by ELISA, AT-Ⅱ cells were used to detect the expression of aquaporin-5and p38MAPK mRNA and protein by reverse transcription-polymerase chain reaction and western blot.Results:1. ELISA showed that the content of TNF-α (pg/ml) in supernatant fluid of the other four groups were markedly elevated,compared with bank control A group,there were statistically significant (P<0.05). Compared with LPS group,the content of TNF-α (pg/ml) in supernatant fluid were rised gradually as the Oxygen concentration increased in different groups(P<0.05). Compared with three different Oxygen concentration groups the content of TNF-α(pg/ml) in supernatant fluid were statistically significant (P<0.05). 2. RT-PCR showed that compared with blank control A group expression of AQP-5mRNA was significantly reduced in the other four groups, the difference was statistically significant (P <0.05).Meanwhile the expression of p38MAPK mRNA were significantly increased, the difference was statistically significant (P<0.05).Compared with group B, C, D, E, expression of AQP-5mRNA decreased gradually,while p38MAPK mRNA expression level gradually increased, the difference was statistically significant (P<0.05); In different Oxygen concentration groups,with increasing oxygen concentration,the expression of AQP-5mRNA decreased gradually, not the same as p38MAPK mRNA expression level which was gradually increased.and there were statistically significant (P<0.05).3. Western blot showed that compared with blank control A group expression of AQP-5protein was significantly reduced in the other four groups, the difference was statistically significant (P <0.05).Meanwhile expression of p38MAPK protein were significantly increased, the difference was statistically significant (P<0.05).Compared with group B, C, D, E, AQP-5protein expressed decreased gradually,while the expression of p38MAPK protion gradually increased, the difference was statistically significant (P<0.05); In different Oxygen concentration groups,with increasing oxygen concentration,AQP-5protein expressed decreased gradually, but the p38MAPK protion expression level gradually increased.and there were statistically significant(P<0.05).Conclusion:In the cell of acute lung injury induced by LPS, the expression of AQP-5mRNA and protein in AT-Ⅱ cells can reduce as the oxygen concentrations increased between40%and90%, and the mechanism might be related to intracellular P38MAPK expression upregulation. |
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