Heat shock protein-70 and 4-hydroxy-2-nonenal adducts in human placental vittous tissue of normotensive, preeclamptic and intrauterine growth restricted pregnancies
Heat shock protein-70 and 4-hydroxy-2-nonenal adducts in human placental vittous tissue of normotensive, preeclamptic and intrauterine growth restricted pregnancies
复制标题
DOI：
10.1016/j.ajog.2005.01.059
复制
发表时间：
2005-09-01
影响因子：
9.8
通讯作者：
Myatt, L
中科院分区：
文献类型：
--
作者：
Hnat, MD;Meadows, JW;Myatt, L
目的：比较正常血压、子痫前期和宫内生长受限(IUGR)孕妇胎盘绒毛组织中氧化应激标志物热休克蛋白-70(HSP70)和脂质过氧化标志物4-羟基-2-壬醛加合物(HNE)的免疫组织化学表达。研究设计：收集正常妊娠(n=5)、妊娠合并子痫前期(n=5)、IUGR(n=5)和子痫前期合并胎儿生长受限(n=4)的胎盘。用Vectaain Elite ABC试剂盒对冰冻切片进行多克隆抗HSP70和抗HNE单抗免疫组化染色。以正常兔血清或小鼠血清作为阴性对照。三名独立的观察者对组织的身份视而不见，检查了每一张载玻片，以确定细胞的定位和免疫染色的强度。结果：各组合体滋养细胞、绒毛外滋养细胞、血管平滑肌和内皮细胞胞浆均有HSP70和HNE加合物的免疫阳性染色。HSP70和HNE加合物的表达被报告为观察者的平均染色强度。总体而言，卡帕在观察员之间表现出了良好的一致性。除了先兆子痫组血管内皮细胞HNE加合物的免疫染色强度明显高于IUGR组(P=0.02)，而IUGR组HSP70的免疫染色强度显著低于IUGR组(P=0.013)外，每组所有组织类型的免疫染色强度均相似。免疫组织化学结果显示，子痫前期和IUGR患者胎盘血管内皮细胞氧化应激和脂质过氧化均有局部改变。然而，在研究组织匀浆时，这些变化被掩盖了。(C)2005 Elsevier Inc.保留所有权利。
Objective: The purpose of this study was to compare immunohistochemical expression of heat shock protein-70 (hsp70), a marker for oxidative stress, and 4-hydroxy-2-nonenal adducts (HNE), a marker for lipid peroxidation, in placental villous tissue of normotensive, preeclampsia, and intrauterine growth restricted (IUGR) pregnancies.Study design: Placentas were collected and flash frozen in liquid nitrogen after delivery from normotensive pregnancies (n = 5), and pregnancies complicated by preeclampsia (n = 5), IUGR (n = 5), and preeclampsia plus IUGR (n = 4). Cryosections were cut and immunostained with polyclonal anti-hsp70 and monoclonal anti-HNE antibodies using Vectastain Elite ABC kit. Normal rabbit serum or mouse IgG were used as negative controls. Three independent observers, blinded to identity of tissue, examined each slide to identify cellular localization and intensity of the immunostaining. Western blot analysis and scanning densitometry were used to quantify and compare the amount of hsp70 and HNE adducts present in tissue homogenates.Results: Positive immunostaining for both antibodies was observed in cytoplasm of syncytiotrophoblasts, extravillous trophoblasts, vascular smooth muscle, and endothelial cells for all groups. Expression of hsp70 and HNE adducts was reported as observers' mean stained intensity. Overall, kappa showed good agreement between observers. Immunostaining intensity was similar in all tissue types for each group with the exception that immunostaining was significantly more intense in the vascular endothelium of the preeclamptic group for HNE adducts (P = .02) and significantly less intense in the IUGR group for hsp70 (P = .013). Scanning densitometric analysis of the Western blots showed no significant difference in total hsp70 and HNE adducts expression in all 4 tissue groups.Conclusion: Immunohistochemistry showed local changes for oxidative stress and lipid peroxidation in the vascular endothelium from placentas of preeclamptic and IUGR pregnancies. However, these changes were masked when studying tissue homogenates. (c) 2005 Elsevier Inc. All rights reserved.