Comparative Proteomic Analysis In Deciduas Of Preeclampsia

Pre-eclampsia,a common type of hypertensive disorder complicating pregnancy,which is a kind of idiosyncratic disease in pregnancy,has been considered as one of the most significant reasons for the death of pregnant women and perinatal babies.Early onset severe pre-eclampsia(ES-PE)is that pre-eclampsia onsets before 34 weeks' gestation while late onset severe pre-eclampsia(LS-PE)onsetting after 34 weeks' gestation.The mechanisms responsible for the pathogeneses of pre-eclampsia are unclear and it has no ideal markers.Recent research has found that there are significantly different in the time of the pre-eclampsia occurrence,mechanism of pre-eclampsia occurrence, clinical,organic hurt,clinical treatment and prognosis between ES -PE and LE-PE.Proteomics is a new technology,which includes protein separation,identification and bioinformatics.It has the virtue of high output and high sensitivity.Human deciduas are a major component of maternal-fetal interface,which contact directly with fetal trophoblast and participate in forming maternal fetal immune to tolerance and maintaining pregnancy.Using technology of proteomics,many scholars have studied peripheral blood,placenta tissues,trophoblast,cerebrospinal fluid and amniotic fluid.However proteomics research on pre-eclamptic deciduas has never been reported.Given this,we studied total proteins of deciduas by using proteomics technology in this study,by comparing the differences of protein expression in deciduas of ES-PE,LS-PE and normal term pregnant women in hopes of finding associated protein of ES-PE and LS-PE,so as to elucidate the pathogenic mechanism of preeclampsia and find out specific biomarkers.Chapterâ… :Separation and identification of differential proteins in deciduas of pre-eclampsiaObjective:To investigate comprehensively differential proteins expressive profiles in deciduas of full-term pregnancy women,early onset severe pre-eclampsia patients,late onset severe pre-eclampsia patients in order to scan out preeclampsia-associationed proteins.Methods:The deciduas from 6 normal women of full-term pregnancy,6 early onset severe pre-eclampsia patients and 6 late onset severe pre-eclampsia patients were collected to obtain the total proteins. The proteins from 3 groups underwent two-dimensional polyacrylamide electrophoresis(2-DE)and coomassie brilliant blue stained gel imaging with PDQuest image analysis software.The peptide mass fingerprinting(PMF)was acquired by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS).The resulted data were then searched against protein data bases for the identification of the proteins,and an initial analysis of identified 29 proteins was made accordingly.Results:1.The well-resolved,reproducible 2-DE patterns of the deciduas of normal full-term pregnancy group,ES-PE group and LS-PE group were acquired.According to them,(425±16)protein sports were seen in normal pregnancy group,(408±10)in ES-PE group and(436±13)in LS-PE group,respectively.2.Image analysis with PDQuest software,proteins sports whose expression difference was at least two times were regarded as significant differential expressional proteins.29 proteins were successfully identified.3.The differential expressed proteins could be divided into ten main groups based on their functions:(1)proteins related to cellur structure (chloride intracellular channel protein 1,F-actin capping proteinα₁ subunit,actin beta,Keratin 19,Vimentin,Myozenin-2,phosphatidylethanolamine binding protein);(2)proteins relative to sigal transduction(phosphogly cerate kinase[2.7.2.3],Annexinâ…¤,Annexinâ…£,Rho-GDP-dissociation inhibitor 1,Rho-GDP dissociation inhibitor 2); (3)energy and metabolic enzymes(protein disulfide isomerase [EC5.3.4.1],ATP synthase D chain,cytide deaminase,glutathione transferase[EC2.7.2.3]);(4)calcium-binding proteins(sorcin,transgelin 2,Tropomyosinα4 chain);(5)other protein(fibrinogen double fragment,fibrinogen beta chain precursor,hemoglobin beta chain,proteasome endopeptidase complex[3.4.5.1]p42 chain);(6)antioxygen enzyme (superoxide dismutase);(7)chaperones(endoplasmin precursor);(8) transport protein(prohibition);(9)preventation protein(DJ-1 protein,α₁-antitrypsin);(10)unknown protein(coiled-coil domain-containing protein 128).Conclusion:1.2-DE patterns of the deciduas of ES-PE group and LS-PE group were successfully established and 29 differential proteins related to the preeclampsia were successfully identified,which had made up the blank of this field.2.Many proteins related to energy and metabolic enzymes and many proteins related to the the function of blood coagulating were found differential expression between normal term pregnancy and preeclampsia, which suggested hypoxidative stress of maternal-fetal interface and the hypercoagulability state might play Crucial roles in the pathogenesis of preeclampsia.3.It was the first time to adopt proteomics technology to study early onset severe preeclampsia and found that there were differential proteins between ES-PE and LS-PE,which suggested ES-PE perhaps have different pathogenic mechanism.Chapterâ…¡:Differential expression of Annexinâ…¤and Rho-GDI in deciduas of each group and its significianceObjective:To confirm the proteomic results,detecting the expression of Annexinâ…¤and Rho-GDI in deciduas of each group and to explore its clinical significiance.Methods:The deciduas were collected from diagnosed clinically with early onset severe preeclampsia(ES-PE:n=10),late onset severe preeclampsia(LE-PE:n=10),and normal term pregnancy(CRL:n=10). Real time quantitative RT-PCR,Western blot and immunohistochemistry technology were preformed to detect expression of Annexinâ…¤and Rho-GDI.Results:1.The results of real time PCR,Western blot and immunohistochemistry were showed as follows:the mRNA and protein expression of Annexinâ…¤was the highest in normal pregnancy,LS-PE group was in the middle level and ES-PE group was the lowest.There was significant difference among three groups(P＜0.05).2.Expression of Rho-GDI mRNA and protein in deciduas of normal women of full term-pregnancy group was highest among three groups,ES-PE group was in the middle level and LS-PE group was the lowest.There was significant difference among three groups(P＜0.05).Conclusion:1.The differential expressional level of annexinâ…¤and Rho-GDI in deciduas of each group was paralleled with the results of proteomics.2.The lower expression of annexinâ…¤in the deciduas of preeclampsia might participate in the hypercoagulability state in PE patients.3.The lower expression of Rho-GDI in the deciduas of preeclampsia might play an important role for pathogenesis of preeclampsia.Chapterâ…¢:Research of differential expression ofα₁-antitrypsin in deciduas and the concentration ofα₁-antitrypsin in maternal peripheral blood of preeclampsiaObjective:To study the differential expression ofα₁-antitrypsin in deciduas of each group and to investigate the change ofα₁-antitrypsin in maternal peripheral blood of preeclampsia and to explore its clinical significiance.Methods:The patient groups and normal group were split as the same as Partâ… .Real time RT-PCR,Western blot and immunohistochemistry methods were used to detect the expression ofα₁-antitrypsin in the deciduas of each group.ELISA method was adopted to detect the concentration ofα₁-antitrypsin in 20 cases maternal peripheral blood of each group.Results:1.Expression ofα₁-antitrypsin in deciduas of normal term pregnancy women was the highest among three groups,the ES-PE group listed in the middle and LS-PE group was the lowest,there was significant difference among three groups(P＜0.01).2.The concentration ofα₁-antitrypsin in the normal pregnancy maternal peripheral blood was the highest(1.85±0.15g/L),ES-PE group in the middle(0.77±0.14g/L)and LS-PE group was the lowest (0.42±0.07 g/L),there was significant difference among three groups(P＜0.01).Conclusion:1.The differential expressional level ofα₁-antitrypsin in deciduas of each group was paralleled with the results of proteomics.2.The lower expression ofα₁-antitrypsin might play a crucial role for the pathogenesis of preeclampsia.3.The concentration ofα₁-antitrypsin in maternal peripheral blood decreased in preeclamptic patients.Perhapsα₁-antitrypsin would be a candicated biomarker of preeclampsia..