SB203580对氧化应激下人绒毛外滋养细胞生物学功能的影响

Effects of SB203580 on biological functions of human extravillous trophoblast cells under oxidative stress

目的·探讨p38丝裂原活化蛋白激酶(p38 MAPK)抑制剂SB203580对缺氧/复氧条件下人绒毛外滋养细胞生物学功能的影响。方法·选取体外培养的早孕绒毛外植体和人绒毛外滋养细胞株HTR8/SVneo,根据干预方式的不同各分为4组:对照组、SB203580组(阻断p38 MAPK信号通路)、缺氧/复氧组(氧化应激模型体外模拟子痫前期)和SB203580+缺氧/复氧组。观察SB203580对氧化应激下早孕绒毛外植体中绒毛外滋养细胞生物学功能的影响。Western blotting检测HTR8/SVneo中p38 MAPK蛋白表达及磷酸化水平,Transwell实验观察细胞侵袭和迁移能力,明胶酶谱法和ELISA法分别检测细胞培养上清液中基质金属蛋白酶2/9(MMP2/9)的活性及可溶性fms样酪氨酸激酶-1(s Flt-1)和可溶性内皮因子(s Eng)的水平。结果·SB203580可促进氧化应激下早孕绒毛外植体中绒毛外滋养细胞的外生性迁移。缺氧/复氧可使HTR8/SVneo细胞的迁移和侵袭能力下降,p38 MAPK磷酸化水平升高,s Flt-1和s Eng分泌增加;而SB203580可使氧化应激下HTR8/SVneo中p38 MAPK磷酸化水平降低,细胞迁移和侵袭能力提高,细胞培养上清液中MMP2/9活性增加、s Flt-1和s Eng的分泌量减少。结论·SB203580对氧化应激下人绒毛外滋养细胞的生物学功能具有保护作用。

To investigate the effects of a p38 mitogen-activated protein kinase (p38 MAPK) inhibitor SB203580 on biological function changes of human extravillous trophoblast cells induced by hypoxia/re-oxygenation (H/R). Methods · In-vitro cultured early pregnancy villus explants and human extravillous trophoblast cell line HTR8/SVneo were assigned to 4 groups according to different interventions, i.e. control group, SB203580 group (p38 MAPK inhibition), H/R group (simulation of preeclampsia by the oxidative stress model), and SB203580+H/R group. The effects of SB203580 on biological functions of human extravillous trophoblast cells under oxidative stress in early pregnancy villus explants were observed. Protein expression and phosphorylation of p38 MAPK in HTR8/SVneo cells were measured with Western blotting. Cell migration and invasion were observed with Transwell migration and Matrigel invasion assay, respectively. Gelatin zymography was used to measure the activity of matrix metalloproteinase (MMP) -2/9 in supernatant. ELISA was used to detect the levels of soluble fms-like tyrosine kinase-1 (sFlt-1) and soluble endoglin (sEng). Results · SB203580 could promote the exogenous migration of human extravillous trophoblast cells in early pregnancy villus explants under oxidative stress. H/R could decrease the migration and invasion of HTR8/SVneo cells , and increase the phosphorylation level of p38 MAPK in HTR8/SVneo cells and the secretion of sFlt-1 and sEng. SB203580 could increase the activity of MMP2/9 in supernatant and cell migration and invasion, decrease the phosphorylation level of p38 MAPK in HTR8/SVneo cells under oxidative stress and the secretion of sFlt-1 and sEng. Conclusion · SB203580 can protect biological functions of human extravillous trophobalst cells under oxidative stress.