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Abnormality of proliferation and differentiation of the trophoblasts in preeclampsia -Correlation between the cell cycle and oxidative stress-

子痫前期滋养细胞增殖分化异常 -细胞周期与氧化应激的相关性-

基本信息

批准号：
14571557
负责人：
KOYAMA Masayasu
金额：
$ 1.86万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

项目摘要

Manifestation analysis of a cell cycle related proteins of trophoblasts in abnormal pregnancyWe obtained chorionic tissues of molar pregnancy, early normal pregnancy, preterm pregnancy andfull-term pregnancy with informed consents at delivery, and we extracted protein and embedded tissues in conventional paraffin. In normal trophoblastic villous cells of each trimester of pregnancy and hydatidiform mole, we confirmed manifestation of cell cycle related proteins such as PCNA, CDK2, p21, p27, p57 and Cyclin Eby Western blot analysis. PCNA, CDK2 and Cyclin Eproteins were strongly expressed in hydatidiform mole and early gestational trophoblasts at the same level. With the late gestation period, the expression of these proteins was diminished. As for p21 which we showed high manifestation in normal trophoblasts of early pregnancy, and deteriorated in the latter period, its stronger expression was found in hydatidiform mole than early pregnancy trophoblasts. As for p57 which is expressed in … More the normal placenta, the manifestation was not found in hydatidiform mole. As for p27 which manifestation enhanced in normal trophoblasts at the early trimester and getting weaker in the latter period, strong manifestation was found in hydatidiform mole than a late gestational trophoblasts. We investigated the localization of p21, p27, p57 in hydatidiform mole by immunohistochemical analysis and compared its localization with the normal gestational trophoblasts.Analysis of influence of oxidative stress in gestational trophoblastic tissueWe obtained placentae at the time of a full-term pregnancy and preterm labor and extracted RNA, protein, and encapsulated a part of these tissues in paraffin. Accumulation of 4hydroxy-2nonenal (HNE), the catabolite of peroxide lipid produced by oxidative stress was investigated by Immunohistochemical staining and Western blot analysis. As for the trophoblastic tissue obtained from the patient suffering from preeclampsia, accumulationof 1-INEwas remarkable in comparison with trophoblastic tissue without any complication. In the same manner, accumulation of HNE was significantly remarkable in trophoblastic tissue with chorioamnionits. We supplemented HNE in cultured trophoblastic tissue and investigated the expression of cyclooxygenase-2 (COX-2) by RT-PCR and Western blot analysis. COX-2 was expressed at transcriptional and translational level at increased for level at time and concentration dependent manner. Production of Prostaglandin also increased at the time dependent manner. These results indicated that trophoblastic tissue caught oxidative stress produced lipid peroxide catabolite increased and activate the cascade of prostaglandin synthetic system. Less

异常妊娠滋养细胞a细胞周期相关蛋白的表达分析我们收集了葡萄胎妊娠、正常早孕、早产和足月妊娠的绒毛组织，并在分娩时知情同意，提取蛋白，常规石蜡包埋。在正常妊娠各期绒毛细胞和葡萄胎绒毛细胞中，我们通过Western blot分析证实了细胞周期相关蛋白如PCNA、CDK 2、p21、p27、p57和Cyclin E的表达。PCNA、CDK 2和Cyclin E蛋白在葡萄胎和早孕滋养细胞中均呈强阳性表达，且表达水平相同。随着妊娠晚期，这些蛋白的表达减少。而p21在正常早孕滋养细胞中呈高表达，在妊娠后期逐渐减弱，在葡萄胎中的表达较早孕滋养细胞强。至于p57，它表达在 ...更多信息正常胎盘，葡萄胎无此表现。而p27在正常滋养细胞中的表达在妊娠早期增强，在妊娠后期减弱，在葡萄胎中表达较晚。本研究应用免疫组化方法检测了p21、p27、p57在葡萄胎中的定位，并与正常妊娠滋养细胞进行了比较。氧化应激对妊娠滋养细胞组织的影响分析用免疫组织化学染色和Western blot分析方法研究了氧化应激产生的脂质过氧化物的分解产物4羟基-2壬烯醛（HNE）的积累。从子痫前期患者的滋养层组织中，1-INE的积累与无任何并发症的滋养层组织相比是显著的。同样地，在有绒毛膜炎的滋养层组织中，HNE的蓄积也非常显著。我们在培养的滋养细胞组织中加入HNE，通过RT-PCR和Western blot分析来检测环氧化酶-2（考克斯-2）的表达。考克斯-2在转录和翻译水平上表达，表达水平呈时间和浓度依赖性增加。前列腺素的产生也以时间依赖的方式增加。这些结果表明滋养层组织受到氧化应激时产生脂质过氧化物分解代谢产物增加，激活前列腺素合成系统的级联反应。少