TRAIL在子痫前期患者胎盘滋养细胞及人脐静脉内皮细胞中的表达及作用研究

Study on the Expression of TRAIL in Placental Trophoblasts and Umbilical Vein Endothelial Cells of Patients with Preeclampsia

目的 探讨肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor-related apoptosis inducing ligand, TRAIL)在子痫前期(preeclampsia, PE)胎盘滋养细胞HTR8-SVneo及人脐静脉内皮细胞(human umbilical vein endothelial cells, HUVEC)中的表达及作用。方法 收集在作者医院住院并分娩的重度子痫前期(severe preeclampsia, SPE)(SPE组)、轻度子痫前期(mild preeclampsia, MPE)(MPE组)患者胎盘及脐静脉组织各20例,同时收集20例无高血压的正常产妇组织样本作为对照组;利用免疫组织化学法分别检测3组患者HTR8-SVneo细胞及HUVEC中TRAIL的表达,应用Image-ProPlus系统对图片进行分析。采用转染法研究TRAIL对HTR8-SVneo细胞及HUVEC凋亡的影响。结果 免疫组织化学法结果显示:HTR8-SVneo细胞及HUVEC中TRAIL的表达比较,SPE组显著高于MPE组及对照组,同时MPE组高于对照组(P均<0.05)。组内比较,SPE组及MPE组HTR8-SVneo细胞中TRAIL均显著高于HUVEC(P均<0.05);对照组中,两种组织中的TRAIL含量差异无统计学意义(P>0.05)。于HTR8-SVneo细胞中过表达TRAIL后,TRAIL mRNA及其蛋白、凋亡相关蛋白半胱氨酸天冬氨酸蛋白酶-3 (cysteinyl aspartate specific proteinase-3,caspase-3)显著升高(P<0.05);HTR8-SVneo细胞凋亡率显著升高(P<0.05)。将TRAIL转染后的HTR8-SVneo细胞上清液与HUVEC共同培养,利用流式细胞术发现SPE组和MPE组中HTR8-SVneo细胞及HUVEC凋亡率均显著高于对照组(P均<0.05)。结论 PE患者胎盘滋养细胞及HUVEC中TRAIL的表达显著上升,可能是通过介导caspase-3表达,从而诱导PE胎盘滋养细胞及HUVEC凋亡,参与PE的发生及发展。

Objective To investigate the expression and role of tumor necrosis factor-related apoptosis inducing ligand(TRAIL) in placental trophoblast cell HTR8-Svneo and human umbilical vein endothelial cells(HUVEC) in preeclampsia(PE). Methods A total of 20 placental and umbilical vein tissues were collected from patients with severe preeclampsia(SPE) and mild preeclampsia(MPE), and 20 normal maternal tissues without hypertension were collected as control group. The expression of TRAIL in HTR8-SVneo cells and HUVEC was detected by immunohistochemistry. The images were analyzed by Image-Proplus system. The effect of TRAIL on the apoptosis of HTR8-SVneo cells and HUVEC was investigated by transfection method. Results Immunohistochemical results showed that the expression of TRAIL in HTR8-SVneo cells and HUVEC was significantly higher in the SPE group than in the MPE group and the control group, while the MPE group was higher than the control group(all P<0.05). In the intra-group comparison, TRAIL in HTR8-SVneo cells was significantly higher in both SPE and MPE groups than in HUVEC(all P<0.05), while in the control group, the difference in TRAIL content between the two tissues was not statistically significant(P>0.05). After TRAIL overexpression in HTR8-SVneo cells, TRAIL mRNA, TRAIL protein and cysteinyl aspartate specific proteinase-3(caspase-3) protein were significantly increased(P<0.05). The apoptosis rate of HTR8-SVneo cells was significantly increased(P<0.05). The supernatant of TRAIL transfected HTR8-SVneo cells was co-cultured with HUVEC. Flow cytometry showed that the apoptosis rate of HTR8-SVneo cells and HUVEC in the experimental group was significantly higher than that in the control group(all P<0.05). Conclusion The expression of TRAIL was significantly increased in placental trophoblast cells and HUVEC of PE patients, which may be involved in the development and progression of PE by mediating caspase-3 expression and thus inducing apoptosis in PE placental trophoblast cells and HUVEC.