过表达SATB1保护人滋养细胞氧化应激损伤的机制研究

发布时间：2018-12-20 21:23

【摘要】：目的探讨富集AT序列的特异性结合蛋白1(SATB1)参与人绒毛外滋养细胞株(HTR8/SVneo)氧化应激损伤的机制。方法收集2013年9月至2014年9月在重庆医科大学附属第一医院产科行选择性剖宫产术的子痫前期孕妇(n=22)和正常足月妊娠行择期剖宫产术的孕妇的胎盘组织(n=22)。采用免疫组化和Western blotting(WB)检测正常晚孕期及子痫前期胎盘组织中SATB1的表达。构建SATB1过表达慢病毒载体(LV-SATB1)和阴性对照载体(LV-NC)。将滋养细胞分为正常培养对照组、缺氧/复氧(H/R)培养组、LV-SATB1+H/R培养组、LV-NC+H/R培养组。利用免疫荧光和WB法检测SATB1在各组细胞中的表达。采用流式细胞仪检测细胞凋亡指数及细胞内活性氧(reactive oxygen species,ROS)水平。利用Transwell小室模型检测细胞的体外侵袭率。结果 SATB1在子痫前期中的表达显著低于正常晚孕组(0.24±0.02 vs.0.12±0.01,t=9.35,P0.05)。转染LV-SATB1可显著提升H/R培养组的SATB1蛋白水平(0.92±0.17 vs.0.58±0.15,P0.01)。LV-SATB1可抑制H/R导致的细胞凋亡率上升及ROS聚集(P0.01)。H/R组细胞侵袭能力低于正常培养组;上调SATB1的表达可提升H/R组细胞的体外侵袭能力(34.33±10.08 vs.19.33±6.52,P0.05)。结论过表达SATB1可以逆转滋养细胞氧化应激损伤,其有望成为研究子痫前期发病机制的新靶点。
[Abstract]:Objective to investigate the mechanism of oxidative stress injury in extracorporeal trophoblastic cell line (HTR8/SVneo) enriched with AT sequence specific binding protein 1 (SATB1). Methods from September 2013 to September 2014, the placental tissues of preeclampsia pregnant women undergoing selective cesarean section in the first affiliated Hospital of Chongqing Medical University and normal full-term pregnant women undergoing elective cesarean section were collected. Immunohistochemistry and Western blotting (WB) were used to detect the expression of SATB1 in placenta of normal late pregnancy and preeclampsia. SATB1 overexpression lentiviral vector (LV-SATB1) and negative control vector (LV-NC) were constructed. Trophoblast cells were divided into normal culture control group, hypoxia / reoxygenation (H / R) culture group, LV-SATB1 H / R culture group and LV-NC H / R culture group. The expression of SATB1 was detected by immunofluorescence and WB assay. Apoptosis index and intracellular reactive oxygen species (reactive oxygen species,ROS) were measured by flow cytometry. Transwell chamber model was used to detect the invasiveness of cells in vitro. Results the expression of SATB1 in preeclampsia was significantly lower than that in normal late pregnancy group (0.24 卤0.02 vs.0.12 卤0.01 vs.0.12 卤9.35). Transfection of LV-SATB1 significantly increased the level of SATB1 protein in H / R culture group (0.92 卤0.17 vs.0.58 卤0.15). LV-SATB1 could inhibit the increase of apoptosis rate induced by H / R and the aggregation of ROS (P0.01). The invasiveness of cells in H / R group was lower than that in normal culture group. Upregulating the expression of SATB1 could enhance the invasiveness of H- R cells in vitro (34.33 卤10.08 vs.19.33 卤6.52 vs.19.33, P0.05). Conclusion overexpression of SATB1 can reverse oxidative stress injury of trophoblast, and it may be a new target to study the pathogenesis of preeclampsia.
【作者单位】：重庆医科大学附属第一医院产科;重庆医科大学病理科;广州医科大学附属第三医院妇产科;
【基金】：国家自然科学基金(81300509,81300508,81471472)
【分类号】：R714.244

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