The Investigation Of The Effects Of Epo And VEGF In Endometriosis

Endometriosis (EMs) is a frequent disorder that commonly presents with infertility and pelvic pain. Although the precise aetiology of endometriosis is unclear, it is generally considered to involve multiple genetic, environmental, immunological, angiogenic and endocrine processes. Although endometriosis is a benign disorder, recent studies of endometriosis suggest endometriosis could be viewed as a neoplastic process.Erythropoietin (Epo) has been accepted as an multiple functional cytokine,which plays an important role in the metastasis of some tumors. Vascular endothelial growth factor (VEGF) is a typical angiogenesis factor, which may be involved in the angiogenesis process of EMs. Up to now,it has not been reported whether or not Epo and VEGF are involved in endometriosis.AIM: To detect the expression of Epo and VEGF in the level of tissue and cells and their correlation between them, in order to reveal the etiology of endometriosis and quest the effective therapy project.METHODS: Our research work was divided into three parts. The first, detect the protein and mRNAs expression of Epo and VEGF by immunohistochemistry and in situ hybridization.The second, culture, purify and identify ectopic endometrial cells of 12 endometriosis patients and eutopic endometrial cells of IS women in order to draw cell growth curves and prepare for the next research work.The last, put the primary cells into anoxic enviroment, detect the expressions of Epo and VEGF by fluorescent immunohistochemistry , and the concentration of Epo in culture medium by ELISA , and apoptosis rate by FCM. The data was analyzed with SPSS 11.0 statistical software.RESULTS: Our results were spread out below:1. The protein and mRNA expressions of Epo and VEGF were detected in both endometriosis and normal endometrium ,while the expressions in endometriosis are enhansive (P<0.05) .2. There was no difference in the expressions of Epo and VEGF between proliferative phase and secretory phase,respectively, whether in endometriosis or in normal endometrium (P>0.05) .3. In endometriosis, the expression of Epo mRNA was closely correlated with that of VEGF mRNA (C=0.733, P<0.001) .4. Ectopic endometrial cells grew faster than eutopic endometrial cells during log growth phase.5. The expressions of Epo and VEGF in ectopic endometrial cells were higher than in eutopic endometrial cells (P<0.05).6. Hypoxia could lead to enhancement of expressions of Epo and VEGF in ectopic endometrial cells and eutopic endometrial cells, this reinforcement between Epo and VEGF is synergy (C=0.864,7. Both ectopic endometrial cells and eutopic endometrial cells could excrete Epo and the former secretion is higher than the latter. (9.1+ 1.5mIU/ml, 2.5+0.4mIU/ml, q=9.67,P<0.01). Hypoxia could lead to an enhancement of secretion of Epo (14.2+ 1.7mIU/ml, 10.6+1.4mIU/ml).8. Hypoxia could lead to an enhancement of apoptosis both in ectopic endometrial cells and in eutopic endometrial cells(13.1%,24.3%).The administration of 50IU/ml Epo could reduce the apoptosis of ectopic endometrial cells(5.3%).9. Hypoxia could induce an enhancement of the expression of Cleaved caspase-3 both in ectopic endometrial cells and in eutopic endometrial cells (70%, 91%, P<0.05). The administration of 50IU/ml Epo could reduce the expression of Cleaved caspase-3 in ectopic endometrial cells(34%).10. Ultrastructure of ectopic endometrial cells was observed also in anoxic enviroment and found that hypoxia could induce apoptosis and necrosis of ectopic endometrial cells (rate =1 .' 4), while SOIU/ml Epo could decrease the rate (1 : 1.5).CONCLUSION:1. The expressions of Epo and VEGF can be detected in normal endometrium, and there is no difference between the expressions of proliferative phase and the expressions of secretory phase.2. The expressions of Epo and VEGF in ectopic endometrium are obviously enhansive, which may play an important role in the pathogenesis of EMs.3. The exptession of Epo in EMs is closely correlated with that...