Effects Of Free Fatty Acid Mixture On AQP1 Of Pulmonary Microvascular Endothelial Cells Of Rats And Mechanism Of FFA-induced AQP1 Expression

Objective: This study was designed to investigate the effects of free fatty acid mixture on Aquaporin1 expression in pulmonary microvascular endothelial cells of rats and the signaling pathways involved in AQP1 expression regulation.Methods:(1)Culture and identification of rat pulmonary microvascular endothelial cells.(2)Western blotting analysis and Real-time PCR analysis were used to determine protein and mRNA levels of AQP1 after FFA treatment in time-dependent manners in rat PMVECs.Western blotting analysis and Real-time PCR analysis were used to determine protein and mRNA levels of AQP1 after FFA treatment in dose-dependent manners in rat PMVECs.(3)Western blotting analysis was used to detect the activation of p38/JNK/ERK signaling pathways after FFA treatment in rat PMVECs.(4)SB203580 was used to determine the effect of p38 MAPK on FFA-induced AQP1 expression.Results: AQP1 protein expression and mRNA expression increased significantly compared to the blank control group after FFA 6h and 12 h treatment(P<0.01).AQP1 was increased significantly AQP1 protein expression was increased significantly compared to the blank control group after treatment of 200?mol/L FFA and 500?mol/L FFA(P<0.01).AQP1 mRNA expression was increased significantly after treatment of 100?mol/L FFA and 200?mol/L FFA and 500?mol/L FFA(P<0.01).Western blotting analysis showed p-p38 protein expression in FFA group was increased significantly compared to the blank control group(P<0.05).P-JNK and p-ERK protein expression in FFA group had no significance compared to the blank control group(P>0.05).After SB203580 treatment,p-p38 protein expression in FFA+SB203580 group was decreased significantly compared to the FFA group(P<0.01).AQP1 protein expression in FFA group was increased significantly compared to the blank control group(P<0.05).AQP1 protein expression in FFA+SB203580 group was decreased significantly compared to the FFA group(P<0.05).Conclusion: FFA can regulate AQP1 expression in dose-related and time-related manner.FFA can affect the progress of water transport in lung in fat embolism syndrome by increasing AQP1 expression to a certain extent.The mechanism of FFA-induced AQP1 regulation in rat pulmonary microvascular endothelial cells may be relevant to the p38 MAPK signaling pathway.