Studies Of The Correlation Between Vascular Endothelial Growth Factor Soluble Receptor (sflt-1) And Endometriosis Angiogenesis Mechanism

Objective: To study the possible relationship between vascularendothelial growth factor soluble receptor (sflt-1),VEGF,VEGF activity index(VEGF/sflt-1) and angiogenesis in eutopic endometrium from women withand without endometriosis and ectopic endometrium,to explore the role ofangiogenesis inhibitors in angiogenesis of endometriosisMethods：Patients with endometriosis (n=30) accepted operation thespecimen were collected at gynecological department of the the SecondHospital of Hebei Medical University.The age of patients ranged from20to48, according to the American (R-AFs) staging criteria, I-II of15cases, Ⅲ-Ⅳ of33cases.We obtain48parts os ectopic endometrium and32parts ofentopic endometrium.Select the same period of30control endometriums inthe same hospital (proliferative phase18cases,12cases of secretory phase),The age of patients ranged from32to50. All patients were menstrualregularity (28-32days), not combined with other surgical diseases;And theyhad not received hormone therapy six months before operation; withoutpregnancy and breastfeeding history in six months, each group of age was notstatistically differences, all selected cases were pathologically diagnosed.We detect the expressions of VEGF and sflt-1in48ectopicendometrium,32eutopic endometrium and30nomal endometrium byimmunohistchemistry SP method and marked with CD105to measure themicrovascular density (MVD), and the results were analyzed using Image-ProPlus6.0.;after we Using real-time quantitative PCR assay groups ofVEGFmRNA,sFlt-1mRNA expression levels; calculation VEGF activityindex.Results： 1The results of immunohistchemistry:①Immunohistchemistry results showed VEGF and sflt-1is mainly expressedin cytoplasm and membrane of endometrial glandular epithelial cells, stromalcells and vascular endothelial cell in the study groups,and the levels ofexpression in gland epithelial cells is stronger than in the stromal cells. CD105is mainly expressed in the cytoplasm and membrane of the vascularendothelial cells.②Immunohistchemistry results showed that the expression levels of VEGFin eutopic endometrium is significantly higher than ectopic endometrium andnomal endometrium(P<0.01); the expression levels of sflt-1in nomalendometrium is significantly higher than ectopic endometrium and nomalendometrium(P<0.01); And ectopic endometrium, eutopic endometrium andcontrol groups MVD were gradually increased, and pairwise comparisonswere statistically significant (P <0.05).③The expression of VEGF in the secretory phase was significantly higherthan that in the proliferative phase in the control group (P <0.01), but theexpression of sflt-1was significantly higher in the proliferative phase than inthe secretory phase (P <0.01);The MVD in control group is no significantcyclical changes(P>0.05）;VEGF expression was not significant cyclicalchanges in eutopic endometrium (P>0.05), but the expression of sflt-1wassignificantly higher in the proliferative phase than in the secretory phase (P<0.01). MVD was significantly higher in secretory phase than that inproliferative phase (P <0.05).④In the correlation analysis，sflt-1and MVD had obviously a negativecorrelation (r=-0.638, P <0.05) in eutopic endometrium;but there is nocorrelation in ectopic endometrium.In control group VEGF,sflt-1as wellsflt-1and MVD was no significant correlation.2The results of Real-time quantitative PCR:①VEGFmRNA, sflt-1mRNA detected by real-time quantitative PCRstandard curve R2respectively0.99, indicating there is a high degree oflinearity; and the amplification efficiency of over90%, and the melting curve showed a single peak distribution on the specificity of the PCR product isnothing more than an extension increased (Fig.4-6), The is a amplificationcurve peak (Fig.7-9),indicate the experimental primer designed and the genepresent in the substrate.②sflt-1mRNA expression in eutopic endometrium and ectopic endometriumgroup were3.898±1.287,3.278±0.997and1.866±0.916, pairwisecomparisons were statistically significant (P <0.05).VEGFmRNA expressionin ectopic endometrium group, eutopic endometrium and control group were1.961±1.130、2.773±0.854和1.109±0.715，pairwise comparisons werestatistically significant (P <0.05),The VEGF activity index in the controlgroup, eutopic endometrium and ectopic endometrium group values were0.698±0.461,0.918±0.375and0.572±0.397,the value of eutopicendometrium was significantly higher than the ectopic endometrium and thecontrol group (P<0.05),but the ectopic endometrium group was higher, but thedifference was not statistically significant (P>0.05).③There were no menstrual cycle phase-related differences forVEGFmRNA,sflt-1mRNA and VEGF activity index in eutopicendometrium.In the conrol group,the expression of VEGFmRNA in theproliferative phase and the secretory phase were1.425±0.800and2.766±1.096, VEGF activity index in the proliferative phase and the secretoryphase were0.350±0.215and0.905±0.377,It was significantly higher in thesecretory phase than the proliferative phase（P<0.01）。In the conrol group,theexpression of sflt-1mRNA in the proliferative phase and the secretory phasewere4.386±1.400and3.165±0.600， the proliferative phase wassignificantly higher than the secretory phase(P <0.01).④VEGFmRNA expression expression in stage Ⅰ-Ⅱ and Ⅲ-Ⅳstage were1.447±0.749and0.956±0.655，in stage ofⅠ-Ⅱ was significantly higherthan in stage Ⅲ-Ⅳ(P <0.05)；sflt-1mRNA expression expression in stage Ⅰ-Ⅱ and Ⅲ-Ⅳ stage were2.517±0.952and1.570±0.740，in stage ofⅠ-Ⅱwas significantly higher than in stage Ⅲ-Ⅳ(P <0.05)；The expression of forVEGF activity index inⅠ-Ⅱand Ⅲ-Ⅳ stage were0.773±0.649and0.651± 0.351, but the difference was not statistically significant (P>0.05).Conclusion：1VEGF and sflt-1in patients with endometriosis are abnormalexpression in eutopic and ectopic endometrial tissueSflt-1may affect VEGF-VEGFR-sVEGFR axis, leading to the occurrence ofendometriosis.2Sflt-1in endometriosis angiogenesis may play a negative feedbackmechanism of regulation, and possibly by affecting VEGF-VEGFR-sVEGFRaxis trigger the occurrence of endometriosis.