ω-3脂肪酸对人滋养层细胞侵袭和血管生成的影响

Effects of Omega-3 Fatty Acids on the Invasion and Angiogenesis of Human Trophoblast Cells

目的:探讨ω-3脂肪酸对人滋养层细胞(HTR-8/SVneo)侵袭和血管生成的影响。方法:本实验设置了不同浓度二十碳五烯酸(EPA)和二十二碳六烯酸(DHA)处理组,依次为0、1、50和100μmol/L EPA组;0、1、50和100μmol/L DHA组。各组HTR-8/SVneo细胞分别用相应浓度的EPA和DHA培养48 h。然后通过CCK-8法检测细胞增殖,Matrigel Transwell实验检测细胞侵袭。使用EPA和DHA处理的HTR-8/SVneo细胞的上清液培养人脐静脉内皮细胞(HUVEC)6 h,然后检测HUVEC的小管形成能力。通过qRT-PCR和Western blot检测HTR-8/SVneo细胞中三结构域蛋白22 (TRIM22)、信号转导和转录激活因子1(STAT1)、p-STAT1(Tyr701)、基质金属蛋白酶2(MMP2)、MMP9和VEGF的表达。结果:与0μmol/L EPA组或0μmol/L DHA组相比,50μmol/L EPA组、100μmol/L EPA组、50μmol/L DHA组和100μmol/L DHA组的OD450nm、侵袭细胞数量、MMP2和MMP9的蛋白相对表达量均升高(P<0.05)。与0μmol/L EPA组或0μmol/L DHA组相比,50μmol/L EPA组、100μmol/L EPA组、50μmol/L DHA组和100μmol/L DHA组的相对小管长度和VEGF蛋白相对表达量均升高(P<0.05)。与0μmol/L EPA组或0μmol/L DHA组相比,50μmol/L EPA组、100μmol/L EPA组、50μmol/L DHA组和100μmol/L DHA组的TRIM22 m RNA和蛋白相对表达量均升高,而STAT1 m RNA相对表达量和p-STAT1 (Tyr701)蛋白相对表达量均降低(P<0.05)。结论:ω-3脂肪酸处理可促进滋养层细胞的侵袭性和血管生成,其机制可能与TRIM22的上调和STAT1活性的抑制有关。

Objective:To investigate the effects of omega-3 fatty acids on the invasion and angiogenesis of human trophoblast cells(HTR-8/SVneo).Methods:Different concentrations of eicosapentaenoic acid(EPA)and docosahexaenoic acid(DHA)groups were set up:0μmol/L EPA group,1μmol/L EPA group,50μmol/L EPA group and 100μmol/L EPA group;0μmol/L DHA group,1μmol/L DHA group,50μmol/L DHA group and 100μmol/L DHA group.The HTR-8/SVneo cells in each group were cultured with corresponding concentrations of EPA and DHA for 48 h.Then the cell proliferation was detected by the CCK-8 method,and the cell invasion was detected by the Matrigel Transwell test.Human umbilical vein endothelial cells(HUVEC)were cultured for 6 h with the supernatant of HTR-8/SVneo cells treated with EPA and DHA,and then the tubule forming ability of HUVEC was detected.The expression level of tripartite motif-containing 22(TRIM22),signal transducer and activator of transcription 1(STAT1),p-STAT1(Tyr701),matrix metalloproteinase 2(MMP2),MMP9,MMP9 and VEGF in HTR-8/SVneo cells were detected by qRT-PCR and Western blot.Results:Compared with the 0μmol/L EPA group or 0μmol/L DHA group,OD450 nm,number of invaded cells,and relative protein expression level of MMP2 and MMP9 in the 50μmol/L EPA group,100μmol/L EPA group,50μmol/L DHA group and 100μmol/L DHA group were increased(P<0.05).Compared with the 0μmol/L EPA group or 0μmol/L DHA group,the relative tubule length and the relative expression of VEGF protein in the 50μmol/L EPA group,100μmol/L EPA group,50μmol/L DHA group and 100μmol/L DHA group were increased(P<0.05).Compared with the 0μmol/L EPA group or 0μmol/L DHA group,the relative expression levels of TRIM22 mRNA and protein in the 50μmol/L EPA group,100μmol/L EPA group,50μmol/L DHA group and 100μmol/L DHA group were increased,while the relative expression of STAT1 mRNA and p-STAT1(Tyr701)protein were decreased(P<0.05).Conclusion:Omega-3 fatty acid treatment can promote the invasiveness and angiogenesis of trophoblast cells.The mechanism may be related to the up-regulation of TRIM22 and the inhibition of STAT1 activity.