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

异常妊娠滋养细胞细胞周期相关蛋白的表现分析我们在分娩时获得知情同意的磨牙妊娠、早期正常妊娠、早产和足月妊娠的绒毛膜组织，提取蛋白并包埋于常规石蜡中。在妊娠各孕期正常滋养层绒毛细胞和葡萄胎中，我们通过Western blot分析证实了细胞周期相关蛋白如PCNA、CDK2、p21、p27、p57和Cyclin e4的表达。PCNA、CDK2和Cyclin e蛋白在葡萄胎和妊娠早期滋养细胞中均有相同水平的表达。随着妊娠后期，这些蛋白的表达逐渐减少。p21在妊娠早期正常滋养细胞中高表达，在妊娠后期恶化，但在葡萄胎中比妊娠早期滋养细胞表达更强烈。在正常胎盘中表达的p57，在葡萄胎中未见表现。p27在妊娠早期在正常滋养细胞中表现增强，在妊娠后期表现减弱，而在妊娠晚期葡萄胎中表现较强。我们用免疫组化方法研究了p21、p27、p57在葡萄胎中的定位，并与正常妊娠滋养细胞的定位进行了比较。氧化应激对妊娠滋养层组织的影响分析我们在足月妊娠和早产时获得胎盘，提取RNA和蛋白质，并将这些组织的一部分包埋在石蜡中。免疫组化染色和Western blot检测氧化应激产生的过氧化脂质分解产物4 -羟基-2壬烯醛（HNE）的积累情况。对于子痫前期患者的滋养层组织，与滋养层组织相比，1- ine1的积累显著，无任何并发症。同样，在绒毛膜羊膜滋养层组织中，HNE的积累也非常显著。我们在培养的滋养细胞组织中添加HNE，并通过RT-PCR和Western blot分析环氧化酶-2 （COX-2）的表达。COX-2在转录和翻译水平上表达，并呈时间和浓度依赖性水平升高。前列腺素的产生也呈时间依赖性增加。这些结果表明，滋养层组织受到氧化应激后产生的脂质过氧化分解代谢增加，并激活了前列腺素合成系统的级联反应。少

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