Study On The Expression Of VEGF And Its Regulation In Human Endometrium

[Objectives] To establish a model of primary endometrial cell culture from eutopic endometrium. In order to analyze the expression and regulation of vascular endothelial growth factor (VEGF) in human endometrium, approaching the contribution of VEGF in the happening and developing course of endometriosis.[Methods] Selecting eight patients whose hysterectomy will be performed for adenomyoma, to obtain the endometrium when the operation is on. Getting primary cultures of stromal cells from endometrium by techniques of enzymolysis,filtration,adherence and purification,etal. When cells are passaged between 2-5 generations, divide them into two separated groups: normoxia(the differential pressure of O₂ is 19-21Kpa), hypoxia(the differential pressure of O₂ is 8-11Kpa). Exert four kinds of stimulus to every culture medium: estrodiol(E₂,the concertration is 10^-8mol/L), progesterone(P, the concentration is 5×10^-7mol/L) , estrodiol together with progesterone(E₂+P,every concentration is the same)and pure culture medium as blank comparision. After 24 hours, removing the stimulus at the same time, collecting the cells. To detect VEGF mRNA of the cells by the method of real-time fluorescent quantitative RT-PCR.[Results] We succeeded in isolating and culturing 6 samples of eutopic endometrial stromal cells in the 8 ones. VEGF expressed in endometrium is significantly up-regulated by the influence of hypoxia and steroid hormone.Relative expression of VEGF mRNA in stromal cells detected by quantitative RT-PCR :group normoxia, datas from E₂,P,E₂+P and blank comparision are 7.22±1.09,7.92±1.10,7.87±1.13及8.23±1.18 respectively;group hypoxia, the corresponding datas are 6.07±1.04,6.75±1.02,6.19±1.06及9.94±1.34;Contrasting pre-and-post intervention inter-class, VEGF mRNA is increased remarkably(P<0.05). Comparing intra-class, VEGF mRNA is more than that of blank comparision afer stimulated by estrodiol and progesterone(P<0.05). Further, hypoxia combining with steroid hormone will not elevate the expression of VEGF in both cells and supernatants compared to single condition(P>0.05).[Conclusions] It is effective to use our method to obtain endometrial stromal cells as a model of primary endometrial cell culture from eutopic endometrium . Human eutopic endometrium can express VEGF. Hypoxia and Steroid hormone may impact the expression. Hypoxia is a most important influential factor to up-regulate the expression of VEGF in endometrium and would not be effected by simultaneously existed steroid hormone. Either estrodiol or progesterone will up-regulate expression of VEGF. It is possible to annotate a new therapy for endometriosis from a preventative point of view:to decrease the expression of VEGF in endometrium by the regulating factors.