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Alteration in the Expression of Proteins in Unexplained Recurrent Pregnancy Loss Compared with in the NormalPlacenta

机译:与正常人相比无法解释的复发性妊娠损失中蛋白质表达的变化胎盘

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摘要

The placenta is a unique pregnancy-related tissue and plays a key role in occurrence of unexplained recurrent pregnancy loss (URPL). Abnormal placentation might play a key role in occurrence of URPL. Therefore, the purpose of this study was to compare the human placental proteome between URPL placentas and normal placental matched for gestational week. Total placental proteins were extracted, and the two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) technique was used for separation of the placental proteomes. Protein spots differentially expressed between URPL and normal placentas were selected and identified by the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF/TOF) technique after being digested in the gel. Moreover, quantitative real-time PCR and Western blot techniques were used to confirm the differential expression mass results for some differentially expressed proteins. The results indicated that at least 19 protein spots were differentially expressed between URPL and normal placentas (P < 0.05), and twelve of them were successfully identified. While only two proteins were downregulated (calumenin and enolase 1), the remaining ten spots (actin gamma 1 propeptide, cathepsin D prepropeptide, heat shock protein gp96, tubulin beta, tubulin alpha 1, glutathione S-transferase, vitamin D binding protein, prohibitin, actin beta, apolipoprotein A-I) showed increased expression in URPL cases in comparison with normal placentas. Real-time PCR also confirmed the downregulation of calumenin and upregulation of prohibitin and apolipoprotein A-I at the mRNA levels. In conclusion, the results of the present study showed that alteration in the expression of proteins involved in proliferation and migration of endothelial cells as well as control of coagulation by these cells might play an important role in the pathogenesis of URPL.
机译:胎盘是独特的妊娠相关组织,在无法解释的反复妊娠丢失(URPL)发生中起关键作用。胎盘异常可能在URPL的发生中起关键作用。因此,本研究的目的是比较妊娠周匹配的URPL胎盘和正常胎盘之间的人胎盘蛋白质组。提取总胎盘蛋白,并使用二维聚丙烯酰胺凝胶电泳(2D-PAGE)技术分离胎盘蛋白质组。选择URPL和正常胎盘之间差异表达的蛋白斑点,并在凝胶中消化后,通过基质辅助激光解吸/电离飞行时间质谱(MALDI TOF / TOF)技术进行鉴定。此外,定量实时PCR和蛋白质印迹技术被用于确认某些差异表达蛋白质的差异表达质量结果。结果表明,URPL和正常胎盘之间至少有19个蛋白点差异表达(P <0.05),成功鉴定了其中的12个。虽然只有两种蛋白被下调(钙铝蛋白和烯醇酶1),其余十个蛋白点(肌动蛋白γ1前肽,组织蛋白酶D前肽,热激蛋白gp96,微管蛋白β,微管蛋白α1,谷胱甘肽S-转移酶,维生素D结合蛋白,禁止蛋白,肌动蛋白β,载脂蛋白AI)与正常胎盘相比在URPL病例中表达增加。实时PCR还证实了mRNA水平上的钙调蛋白的下调以及禁止素和载脂蛋白A-1的上调。总之,本研究的结果表明,参与内皮细胞增殖和迁移以及这些细胞对凝血的控制的蛋白质表达的改变可能在URPL的发病机理中起重要作用。

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