1－磷酸鞘氨醇对人脐静脉血管内皮细胞增殖和迁移的影响及机制

摘要

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.%目的：观察1－磷酸鞘氨醇（ S1P）对人脐静脉内皮细胞（ HUVEC）增殖和迁移的影响，探讨其作用机制。方法将HUVEC分为A～F组，A组只加细胞培养液，B～F组分别加入10－10、10－9、10－8、10－7、10－6 mol／L的S1P；采用CCK－8法检测各组培养48 h的OD450值，Transwell实验计数各组穿膜细胞数。将HUVEC分为3组，空白对照组加细胞培养液，低浓度组和高浓度组分别加10－7、10－6 mol／L的S1P；采用Western blot法检测ERK、p－ERK蛋白，计算二者比值。结果 A～F组OD450值分别为0．50±0．03、0．51±0．06、0．65±0．04、0．82±0．19、0．99±0．06、0．99±0．02，穿膜细胞数分别为（66．59±9．11）、（76．72±4．80）、（83．75±7．59）、（101．39±5．03）、（101．50±8．96）、（99．38±13．03）个／HP。 D、E、F组与A组比较，P均＜0．05；B、C组与A组比较，P均＞0．05；D、E、F组间比较，P均＞0．05。空白对照组、低浓度组和高浓度组p－ERK蛋白与ERK蛋白比值分别为0．39±0．03、0．94±0．07、0．97±0．05，低浓度组和高浓度组与空白对照组比较，P均＜0．01。结论 S1P可通过MAPK／ERK通路促进HUVEC的增殖和迁移。