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

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

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、10-6mol/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、10-6mol/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。

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.