전자간증 태반에서 프로테오믹스를 이용한 단백질 분석

다국어 초록 (Multilingual Abstract)

Preeclampsia is a major cause of maternal-fetal morbidity and mortality. The causes and pathogenesis of preeclampsia are unknown although many possibilities are being investigated. Recent studies have suggested that abnormal expression of a naturally occurring molecule of placental origin may contribute to the pathogenesis of preeclampsia. Analyzing the proteins expressed in the preeclamptic placenta is essential to understand the causes and pathogenesis of preecalmpsia. Proteomics provides a powerful approach to screen for alterations in protein expression under conditions of preeclampsia.
The objective of this study is to identify the quantitative changes of proteins in the preeclamptic placenta. In this study, twelve paired placental tissues of preeclampsia and control pregnancy were used for proteomic approach. A vertical, two-dimensional polyacrylamide gel electrophoresis (2-DE) system and PDQuest image analysis software were used to provide a high level of between-gel reproducibility and electrophoretic separation to accurately locate each differentially expressed protein. Matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS) and protein databases were used to characterize each differentially expressed protein.
Image analysis of silver stained two-dimensional gels revealed that 60 protein spots showed significantly quantitative variations, of which 14 spots were up-regulated and 46 spots were down-regulated in preeclamptic placenta. Among the 60 protein spots with significant difference, twelve protein were identified by MALDI-TOF-MS. Biological function of these identified proteins are known to be anti-oxidation, metabolism, chaperones, signalling molecules, transport protein and cellular structural Differential expression of some of the 12 identified protein spots was further confirmed by Western blot analyses and immunohistochemistry. The expression pattern of four protein spots - peroxiredoxin II, heat shock protein 27, human chorionic somatomammotropin and vimentin - was analysed. Western blotting analysis showed that peroxiredoxin II and heat shock protein 27 were up-regulated and human chorionic somatomammotropin and vimentin were down-regulated in preeclamptic placenta. The immunohistochemical localization of these four proteins were examined using an avidin-biotin peroxidase complex technique. Immunoreactivity of peroxiredoxin II, heat shock protein 27 and human chorionic somatomammotropin were observed mostly in the trophoblastic layer. Immunoreactivity of vimentin was observed mostly in the intravillous endothelial cells.
These result may serve important resources for subsequent proteomics studies for analyzing the protein variability in preeclamptic placenta tissues and also for the investigation of the molecular basis of preeclampsia pathophysiologic research.

목차 (Table of Contents)

• 1.서론 = 1
• 2.실험 재료 및 방법 = 4
• 3.결과 = 16
• 4.고찰 = 40
• 5.결론 = 46
• 참고문헌 = 48