子痫前期孕产妇胎盘组织中过氧化氢对人类白细胞相关抗原G表达的影响

Effect of hydrogen peroxide on human leukocyte antigen-G expression in placental trophoblasts in pre-eclampsia

目的探讨子痫前期孕产妇胎盘组织中氧化应激产物过氧化氢（H202）对胎盘滋养细胞中的人类白细胞相关抗原G（HLA-G）表达的影响。方法选择2008年10月一2009年10月南京医科大学第一附属医院住院的产妇40例，其中子痫前期组20例，健康妊娠组20例。采用比色法检测两组产妇胎盘组织中H2O2水平；采用蛋白印迹法检测两组产妇胎盘组织中HLA-G蛋白的表达水平；并对胎盘组织中H2O2水平与HLA-G蛋白表达水平进行相关性分析。人绒毛膜癌细胞株JEG-3细胞培养24h后贴壁完全，分为两组，干预组加入浓度为175μmol／L的H2O2，对照组不加H2O2，两组细胞培养24及48h后，采用蛋白印迹法和细胞免疫荧光法检测JEG-3细胞中HLA—G蛋白表达水平和强度的变化。结果（1）子痫前期组产妇胎盘组织中H2O2水平为（105±13）nmol·mg^-1·prot^-1，健康妊娠组产妇胎盘组织中H2O2水平为（62±18）nmol·mg^-1·prot^-1，子痫前期组较健康妊娠组明显升高，且差异有统计学意义（P〈0．05）。（2）子痫前期组产妇胎盘组织中HLA—G蛋白表达水平为0．20±0．08，健康妊娠组为1．67±0．65，子痫前期组比健康妊娠组下降了88％，两组比较，差异有统计学意义（P〈0．05）。（3）两组产妇胎盘组织中H2O2水平与HIJA-G蛋白表达水平呈显著负相关（r=一0．895，P=0．000）。（4）H2O2干预JEG一3细胞24h后，JEG一3细胞中HLA—G蛋白表达水平为1．95±0．25，干预48h后为0．65±0．08；与未干预细胞中的3．21±0．33比较，分别下降了39％和80％，干预24、48h分别与对照组比较，差异均有统计学意义（P〈0．05）；干预48h与干预24h比较，HLA—G蛋白表达水平下降了67％，两者比较，差异有统计学意义（P〈0．05）。H：O：干预JEG一3细胞48h后荧光显微镜下显示，干预组细胞中HLA—G表达强度明显弱于对照组。结论子痫前期产妇胎盘滋养细胞中的高水平H2O2能下调胎盘滋养细胞中HLA—G蛋白的表达，从而参与子痫前期的发病。

Objective To explore the effect of hydrogen peroxide （H202 ） on the expression of human leukocyte antigen-G （HLA-G） in placental trophoblasts in pregnant women with pre-eclampsia Methods Forty pregnant women, delivered through cesarean section in the Department of Obstetrics of and Gynecology, the First Affiliated Hospital of Nanjing Medical University from October 2008 to October 2009, were enrolled, including 20 women with pre-eclampsia and 20 healthy gravidas （control group）. Colorimetry and western blot were applied, respectively, to determine the level of H202 and the expression of HLA-G protein in placental tissues and the correlation between them were analyzed. After 24 hours of seeding, JEG-3 cells （the HLA-G positive cell line of choriocarcinoma） were divided into two groups： intervention group （exposure to 175 tLmol/L H202 ） and control group （without H202 ）. Immunofluorescence and western blot were used to investigate the expression of HLA-G protein in JEG-3 cells at 24 hours and 48 hours after incubation. Results （ 1 ） The level of H2 02 in placenta in the pre-eclampsia group was significantly higher than that in control group [（105 ＋13） nmol mg- prot- vs （62-＋18） nmol mg-1 prot-1, P〈 0.05]. （2） The expression of HLA-G protein in placenta of the pre-eclampsia group was reduced by 88%