The Mechanism Of LXA4 Intervention On The Oxidative Stress Induced By Uric Acid In Human Umbilical Vein Endothelial Cells

Objective:To investigate the effect and mechanism of lipoxin A4(LXA4)on uric acid induced oxidative stress of human umbilical vein endothelial cells(HUVECs).Method: HUVECs had been cultured for the whole experiment. The HUVECs was treated with different concentrations of uric acid(0mg/dL,4mg/dL, 8mg/dL, 12mg/dL, 16mg/dL) for 24 h or with 12mg/dL uric acid for different times(3h, 6h, 12 h, 24 h, 48h), respectively. ROS were detected by a fluorescence probeDCFH-DA. HUVECs was pre-treated with LXA4 of concentrations(0nM, 1nM, 10 nM and 100 nM, respectively) and times(0min,15 min, 30 min, 1h and 2h, respectively) before treated with 12mg/dL uric acid for 24 h. HUVECs was pre-treated by 100 nM LXA4, 10μM DPI and 1μM rotenonerespectively, and then treated by 12mg/dL uric acid for 24 h. The concentration of ROS was detected among these groups. The activity of NADPH oxidase and p47 phoxprotein was measured by Lucigenin enhanced chemiluminescence and Western blot among control, UA, LXA4 and UA+LXA4 groups, respectively.Result: UA induced ROS productions were on time- and dosedependent manners in HUVECs, and the peak of ROS productions was at12mg/dL UA treatment cells for 24 h. Compared to UA group, the concentrations of ROS in LXA4+UA and DPI+UA group were remarkable descended(P<0.05). There was no different between UA and rotenone+UA group(P>0.05).The activity of NADPH oxidase in UA group were increased significantly compared to control group(P<0.05), but it was suppressed inUA+LXA4 group(P<0.05). The protein expression of p47 phox in cytoplasm was lower in UA group than control(P<0.05), but the expression of p47 phox was significantly increase after pretreatment with LXA4(P<0.05). On the contrary, the protein expression of p47 phox in cytomembrane was higher in UA group than control(P<0.05), but the expression of p47 phox was significantly decreased after pretreatment with LXA4(P<0.05).Conclusion: LXA4 inhibit UA induced ROS production in HUVECs.This mechanism might be through inhibiting p47 phox trafficing from cytoplasm to cytomembrane,resulted in inhibiting the activation of NADPH oxidase.