Kristensen Karl, Larsson I, Hansson S R
Department of Obstetrics and Gynaecology, University Hospital, Lund, Sweden.
Mol Hum Reprod. 2007 Mar;13(3):189-95. doi: 10.1093/molehr/gal111. Epub 2007 Jan 16.
Trophoblast invasion is regulated by proteinases and their inhibitors. Cystatin C inhibits cysteine proteinases. The serum concentration of cystatin C is increased in late pregnancy and pre-eclampsia. We aimed to investigate whether the expression of cystatin C is increased in the pre-eclamptic placenta and to investigate the expression pattern of cystatin C mRNA and protein in placental tissue. Tissue samples from the central part of the placenta from 13 normal and 22 pre-eclamptic pregnancies were included. We used real-time polymerase chain reaction (RT-PCR) and in situ hybridization for mRNA expression analysis and immunohistochemistry and Western blotting for protein expression analysis. RT-PCR showed a significantly higher expression of cystatin C mRNA in pre-eclampsia than in normal pregnancy, with the highest expression in cases with severe pre-eclampsia. In situ hybridization revealed a distinct pattern of high expression in the extravillous trophoblast cells of the basal plate and low expression in the syncytiotrophoblast covering villi. The cystatin C protein distribution matched the mRNA expression pattern. Western blot analysis revealed an increased protein expression in cases with severe pre-eclampsia and confirmed the presence of cystatin C in amniotic fluid samples. The high expression of cystatin C mRNA in the extravillous trophoblast cells of the basal plate suggests a role for cystatin C in the regulation of proteases in placentation. Placental expression and secretion of cystatin C could contribute to the elevated maternal plasma levels seen in pre-eclampsia.
滋养层细胞侵袭受蛋白酶及其抑制剂调控。胱抑素C抑制半胱氨酸蛋白酶。妊娠晚期及子痫前期患者血清胱抑素C浓度升高。我们旨在研究子痫前期胎盘组织中胱抑素C的表达是否增加,并研究胎盘组织中胱抑素C mRNA和蛋白的表达模式。纳入了13例正常妊娠和22例子痫前期妊娠胎盘中央部分的组织样本。我们采用实时聚合酶链反应(RT-PCR)和原位杂交进行mRNA表达分析,采用免疫组织化学和蛋白质印迹法进行蛋白质表达分析。RT-PCR结果显示,子痫前期患者胱抑素C mRNA的表达显著高于正常妊娠,重度子痫前期患者表达最高。原位杂交显示,基底板绒毛外滋养层细胞高表达,覆盖绒毛的合体滋养层细胞低表达。胱抑素C蛋白分布与mRNA表达模式相符。蛋白质印迹分析显示,重度子痫前期患者蛋白表达增加,并证实羊水样本中存在胱抑素C。基底板绒毛外滋养层细胞中胱抑素C mRNA的高表达表明,胱抑素C在胎盘形成过程中对蛋白酶的调控发挥作用。子痫前期患者母体血浆中胱抑素C水平升高可能与胎盘表达和分泌胱抑素C有关。