Effect of sphingosine-1-phosphate on migration and proliferation of HUVECs and its mechanism

YUAN Yong-hui; QI Xun; YANG Fang

doi:10.3969/j.issn.1002-266X.2015.34.004

Abstract：Objective To observe the effect of sphingosine-1-phosphate ( S1P) on migration and proliferation of hu-man umbilicus vein endothelial cells ( HUVECs) and to investigate its mechanism.Methods HUVECs were divided into, group A to group F, group A was only added with cell culture medium, group B to group F were added with different con-centrations of S1P (10 -10, 10 -9, 10 -8, 10 -7 and 10 -6 mol/L, respectively).CCK-8 method was used to detect the OD450 value 48 h after treatment, Transwell cell migration assay was employed to investigate the number of migrated cells.HU-VECs were divided into three groups: the blank control group which was only added with cell culture medium, low-dose group and high-dose group were added with 10 -7 and 10 -6 mol/L S1P;the protein expression levels of phospho-ERK and ERK were detected by Western blotting, then the ratio of pERK to ERK was calculated.Results The OD450 values of group A to group F were 0.50 ±0.03, 0.51 ±0.06, 0.65 ±0.04, 0.82 ±0.19, 0.99 ±0.06 and 0.99 ±0.02, and the numbers of migrated cells were (66.59 ±9.11), (76.72 ±4.80), (83.75 ±7.59), (101.39 ±5.03), (101.50 ± 8.96) and (99.38 ±13.03) /HP.Significant difference was found between group A and groups D, E, F ( all P<0.05);no difference was found between group A and groups B, C (all P>0.05), and no difference was found between groups D, E and F (all P>0.05).The ratios of pERK to ERK in the blank control group, low-dose and high-dose groups were 0.39 ±0.03, 0.94 ±0.07 and 0.97 ±0.05, and significant difference was found between the blank control group and low-dose group, high-dose group (all P<0.01).Conclusion S1P can promote the migration and proliferation of HU-VECs through MAPK/ERK signaling pathway.