The Study Of The Stromal Cells' Culture From Women With Endometriosis In Vitro And GnRH Ⅰ,GnRH Ⅱ Inhibits Its Proliferation

ObjectiveTo culture the eutopic and ectopic endometrial stromal cells from patients with endometriosis and identify them in vitro,then different concentrations of GnRHⅡwere added in the stromal cells in vitro culture, and compared to GnRHⅠ,to investigate the direct effect of GnRHⅡand GnRHⅠon the endometrial stromal cell in vitro,to compare and analyse them.MethodsEutopic and ectopic endometrium were obtained from thirty patients who underwent gynaecological surgery for endometriosis.They did not take hormone in past six months,and had no other gynecologic,endocrine secretion diseases,immune and metabolismic diseases. Endometrial tissues were collected by cerettage at the time of surgery in women with ovarian endometrioma,endometrial tissues were collected from the walls of endometriomas.The stromal cells were separated from the grandular epithelium and cultured,identificated in vitro,which were divided into three groups:1.treated by 10^-10,10^-8,10^-6M GnRHⅡ; 2.treated by 10^-10,10^-8,10^-6M GnRHⅠ;3.control group,not treated by GnRH,and then were continued to culture for 24 hours,48 hours,72hours.MTT was used to measure survival cell,and calculated to compare the rate of stromal cells' inhibition in vitro.Results1.Eutopic endometrial stromal cells of survival rate with the method of trypsin,collagenase,mesh filtration and centeifugation was 83.33%,ectopic endometrial stromal cells of survival rate is 66.67%.2.The eutopic endometrial stromal cell was cultured and treated with graded concentrations of GnRHⅡ(10^-10M,10^-8M,10^-6M) for 24 hours,cell inhibition rate(%)were(15.32±2.43,26.41±2.75,38.06±4.15) respectively,for 48 hours(%)were(30.41±3.50,41.78±4.49,53.34±5.83) respectively,for 72 hours(%)were(50.01±3.70,63.29±4.47,81.58±3.44) respectively.On the ectopic endometrial stromal cell,for 24 hours(%)were (38.42±2.67,51.35±3.70,62.84±4.13)respectively,for 48 hours(%)were (53.41±3.79,64.47±4.78,76.39±5.71)respectively,for 72 hours(%)were (70.29±2.87,83.25±3.73,93.39±4.85)respectively,the cell inhibition rate of the same endometrial stromal cell with different graded concentrations of GnRHⅡat the same time was gradually increased,significant difference was observed(P＜0.05),the cell inhibition rate of the same endometrial stromal cell with same concentrations of GnRHⅡat different time was gradually increased,significant difference was observed(P＜0.05), in dose-and time-dependent manner,significant difference was observed (P＜0.05),the cell inhibition rate of ectopic endometrial stromal cell was higher than that of eutopic endometrial stromal cell,significant difference was observed(P＜0.05).3.The eutopic endometrial stromal cell was cultured and treated with graded concentrations of GnRHⅠ(10^-10M,10^-8M,10^-6M) for 24 hours,cell inhibition rate(%)were(5.42±0.93,11.19±1.29,23.97±3.00)respectively, for 48 hours(%)were(18.00±2.02,28.98±3.91,41.98±4.86)respectively, for 72 hours(%)were(32.20±2.91,46.94±4.08,60.73±4.70)respectively. On ectopic endometrial stromal cell,for 24 hours(%)were(25.63±1.24, 37.53±2.35,48.70±3.15)respectively,for 48 hours(%)were(38.54±2.77, 50.70±3.46,60.65±4.25)respectively,for 72 hours(%)were(58.38±1.36, 69.75±2.18,78.69±3.75) respectively,the cell inhibition rate of the same endometrial stromal cell with different graded concentrations of GnRHⅠat the same time was gradually increased,significant difference was observed.(P＜0.05),the cell inhibition rate of the same endometrial stromal cell with same concentrations of GnRHⅠat different time was gradually increased,significant difference was observed(P＜0.05),in dose and time dependent manner,significant difference was observed(P＜0.05), the cell inhibition rate of ectopic endometrial stromal cell was higher than that of eutopic endometrial stromal cell,ignificant difference was observed(P＜0.05).4.GnRHⅡhad higher cell inhibition rate on endometrial stromal cell in vitro than GnRHⅠ,significant difference was observed(P＜0.05).Conclusion 1.Highly purified eutopic and ectopic endometrial stromal cells were seperated and cultured successfully with the method of trypsin,collagenase,mesh filtration and centeifugation,providing a successful cell model for EMs,providing expremental basis for the pathogenesis and drug therapy of EMs.2.The successful culture of ovarian stromal cells for EMs was its site.3.GnRHⅡhad more antiproliferative effects on endometrial stromal cell than GnRHⅠanalogues(goserelin) in vitro,especially on ectopic endometrial stromal cells,it suggested GnRHⅡwas more effective than GnRHⅠ,providing a new theory for finding new drug for EMs.