孕妇血清及胎盘组织中晚期糖基化终末产物及其受体的表达水平变化与子痫前期发病的相关性

Correlation of the expressions of advanced glycation end products and its receptor in serum and placenta with the pathogenesis of preeclampsia

目的探讨孕妇血清及胎盘组织中晚期糖基化终末产物（AGE）及晚期糖基化终末产物受体（RAGE）的表达水平变化与子痫前期发病的相关性。方法选择2013年12月至2014年6月在青岛大学附属医院产科住院的重度子痫前期孕妇32例为重度子痫前期组，选取同期晚期妊娠健康孕妇30例为健康对照组，两组孕妇均为剖宫产术分娩。采用ELISA法检测孕妇血清中AGE、可溶性晚期糖基化终末产物受体（sRAGE）、肿瘤坏死因子α（TNF-α）及胎盘组织中AGE、TNF—α的水平；免疫组化SP法检测胎盘组织中AGE、RAGE蛋白的表达定位；实时荧光定量PCR技术检测胎盘组织中RAGE和TNF-α mRNA的表达水平；蛋白印迹法（western blot）检测胎盘组织中AGE、RAGE及TNF-α蛋白的表达水平。结果（1）重度子痫前期组孕妇血清中AGE、sRAGE及TNF-α水平分别为（538±75）、（367±86）及（322±40）ng，L，均显著高于健康对照组的（454±50）、（286±35）及（270±35）ng，L，两组分别比较，差异均有统计学意义（P〈0．05）。重度子痫前期组孕妇血清AGE水平与TNF．d水平呈显著正相关（r=0．588，P〈0．05），sRAGE水平与TNF-α水平无相关性（r=0．041，P〉0．05）。（2）重度子痫前期组胎盘组织中AGE及TNF-α水平分别为（500±82）及（334±57）ng，L，明显高于健康对照组的（431±74）及（263±46）ng／L，两组分别比较，差异均有统计学意义（P〈0．05）。重度子痫前期组孕妇胎盘组织中AGE水平与TNF．仪水平呈显著正相关（r=0．406，P〈0．05）。（3）重度子痫前期组及健康对照组胎盘组织中AGE及RAGE蛋白均主要定位表达于胎盘合体滋养细胞、巨噬细胞及血管内皮细胞，AGE主要定位于细胞质内，RAGE主要定位于细胞膜和细胞质内。（4）重度子痫前期组胎盘组织中RAGE及TNF—dmRNA的表达水平分别为12．6±4．6及10．4±2．4，均高于健康对照组的0．9±0.4及3．5±0．9，两组分别比较，差异均有统计学意义（P〈0．01）。（5）重度子痫前期组胎盘组织中AGE、RAGE及TNF-α蛋白的表达水平分别为0．68±0．06、0．82±0．08及0．76±0．08，均显著高于健康对照组的0．46±0．05、0．42±0．09及0．52±0．07，分别比较，差异均有统计学意义（P〈0．01）。结论重度子痫前期孕妇血清及胎盘组织中AGE及RAGE表达水平明显高于健康孕妇，同时伴有胎盘组织炎性因子TNF—α水平的明显升高，提示AGE及RAGE可能通过诱发孕妇全身和胎盘局部炎性反应参与子痫前期的发病过程。

Objective To investigate the correlation of the expressions of advanced glycation end products（AGE） and the receptor for advanced glycation end products（RAGE） in serum and placenta with the pathogenesis of preeclampsia. Methods From December 2013 to June 2014, 32 women with severe preeclampsia who received cesarean section in the Affiliated Hospital of Qingdao University were recruited in the study, defined as the severe preeelampsia group. 30 healthy pregnant women who received cesarean section in the same hospital were recruited as the control group. ELISA was used to measure the maternal serum AGE, soluble receptor for advanced glycation end products （sRAGE） and tumor necrosis factor-α （TNF-α） in these women. Furthermore, ELISA was also used to measure AGE and TNF-α in the placenta. The localizations of AGE and RAGE protein in placentas were detected by immunohistochemical SP method. RAGE and TNF-α mRNA expression in placentas were measured by real-time quantitative PCR. AGE, RAGE and TNF-α protein expression in placentas were measured by western blot, respectively. Results （1） The serum levels of AGE, sRAGE and TNF-α in the severe preeclampsia group were （538 ± 75）,（367 ± 86） and （322 ± 40） ng/L,respectively. They were significantly higher than those in the control group[（454 ± 50）, （286 ± 35） and （270 ± 35） ng/L, respectively] （P〈0.05）. The levels of AGE showed positive correlation with the levels of TNF-α（r=0.588,P〈0.05）,while the levels of sRAGE showed no correlation with TNF-α（r=-0.041, P〉0.05）. （2） In the severe preeclampsia group, the levels of AGE and TNF-α in placentas were （500 ± 82） and （334± 57） ng/L, which were higher than those in the control group [（431 ±74） and （263 ± 46） ng/L, respectively] （P〈0.05）. The levels of AGE showed positive correlation with the levels of TNF-α （r=0.406,P〈 0.05）. （3）AGE and RAGE protein mainly located in the syncytiotrophoblasts, macrophages and vascular endothelial cells in the placentas of the two groups. AGE expressed mainly in the cytoplasm, and RAGE expressed in the cytoplasm and cell membranes. （4） RAGE and TNF-α mRNA expression in the placentas of the severe preeelampsia group were 12.6 ± 4.6 and 10.4 ± 2.4, which were significantly higher than those in the control group （0.9 ± 0.4 and 3.5 ± 0.9,P〈0.01）. （5） The expressions of AGE ,RAGE and TNF-α protein in placentas of the severe preeclampsia group were 0.68 ±0.06, 0.82 ± 0.08 and 0.76 ± 0.08. All were significantly higher than those of the control group （0.46 ± 0.05,0.42 ± 0.09 and 0.52 ± 0.07;P〈0.01）. Conchtsions The levels of AGE and RAGE in serum and placentas elevated in the severe preeclampsia group, and the expression of TNF-α also elevated. These indicated that AGE and RAGE might be involved in the systemic inflammatory response and local inflammatory response in placentas, and then caused the preeclampsia.