| Background:Endometriosis (EM) is a frequently-occurring disease in women of childbearing age, and ofgen causes pelvic mass, dysmenorrhea and infertility, which affecting reproductive health and quality of life of women. Syndrome is the comprehensive performance of etiology, location, quality and trend of disease, and it’s the central conception of traditional Chinese Medicine, and to reveal the scientific essence of syndrome is the crucial part of modernization of TCM. Blood stasis syndrome is common type in clinic including gynecology. Prof. Wei thinks that damp-blood stasis is the crucial pathegenesis of endometriosis. Proteomics has been one of the most important approaches in the researches of pathogenesis, biomarker screening, target to pharmacological action and gene interference,characterized by the analysis of global protein changes in certain tissue or cell in integrity and high flux.Objective:①To separate the global protein in3types with blood stasis syndrome of eutopic endometrium in women with endometriosis and normal endometrium tissues by2-DE and analyze the differential protein in2-D profiles, and identify them with MS.②To detect the expression of VEGF,Ang-2,Tie-2,survivin,PCNA,Livin,Cyclin D1,P16,MIF an Foxp3in the eutopic endometrium of blood stasis syndrome by immunohistrochemistry.③To discover the essence of blood stasis syndrome by proteomics and to build the differentiation and diagnosis system of blood stasis in endometriosis, and to find the valuable biomarker and target.④To bulid the protein database of TCM syndrome and serve the externalize and standardization of differentiation, to improve the accuracy of differentiation and effect.Methods:The globel proteins of6endometrium tissue specimens of blood stasis syndrome in endometriosis and the normal endometrium were extracted and separated by2-DE.The2-DE image profiles were analyzed and measured by PDQuest7.1.1including cut, spot detection.These differential expressing spots were cut for in-gel enzymolysis digestion, and the peptides were identified by MALDI-TOF-TOF/MS and the data were searched in protein database. Those proteins with stable and significant differential expression were selected as candidates to blood stasis syndrome and endometriosis associated protein. The differential expression of VEGF、Ang-2、Tie-2、MIF、survivin、PCNA、Livin、Cyclin D1、P16、 Foxp3were detected by immunohistrochemical methods, and mesured the IOD values by Imagepro-Plus5.0and analyzed the difference by statistics.Results:①6couples of2-DE images with high resolution about blood stasis syndrome with women endometriosis were obtained,66spots were selected for obvious diffential expression.22proteins were identified by MS and database search.The function and cell location of those proteins were classified.②According to the six groups of compare of those10indexes expression in three types of blood stasis syndrome of endometriosis, there were four groups results with significant differencial expression of Ang-2,MIF,Foxp3;three groups of VEGF with significant differencial expression; two groups of Livin,PCNA,P16with significant differencial expression; one group of Cyclin D1,Tie-2with significant differencial expression; there were on different expression of survivin.Conclusion:①By2-DE and MS, the differential expressed endometriosis-associated proteins of blood stasis syndrome and health women were separated and identified. Those differential proteins might be the candidate biomarkers of endometriosis. And there were different expression of eutopic endometrium of three types of blood stasis syndrome, which might be the origin of essence of syndrome.②Ang-2,Foxp3,MIF might be the potential differentiational biomarkers of blood stasis syndrome;Survivin was unsuited as the candidate for diagnosis and differentiation. |
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