| Objection1.Explore the differences of BMP-15 content in follicular fluid between endometriosis infertile patients and just male severe less or weak sperm infertility or without hydrosalpinx infertile patients.How the BMP-15 influence the reproduction.2.To evalute how the apoptosis rate of granulocyte influence the oocytes quality.Methods1.choose the infertile patients in reproductive center of Sir Run Run Shaw Hospital College Of Medicine Zhjiang University from June 2009 to november 2010. We put the patients that diagnosed as endometriosis after laparoscopic or ovarian cyst puncture into the experimental group, just male severe less or weak sperm infertility or without hydrosalpinx patients are control group members.2.All patients we chosed recorded their age,eggs,fertilization number,normal embryo,good embryo number.we statistic their pregnancy rate.3.We collect the follicular fluid without blood into EP tube when ovum pick up.We make the follicular fluid rotating to remove such as cell visible part.Adopting Western blot method to determine the content of BMP-15 in follicular fluid between the infertile patients with endometriosis (EMS) and the patients with tube sexual factors or the man factors. 4.We return the granulosa cells around the maturation oocytes into the EP tube and remove the fluid.We put hyaluronidase into the EP tube. Using the FITC and PI coloured the granulosa cells,then make the granulosa cells away from the light.Using the flow cytometry analysis apoptosis rate of granulosa cell of two groups patients.5 We return the granulosa cells around the maturation oocytes into the EP tube and remove the fluid.Conserving the granulosa cells in the-80℃freezer.When we begin to do experiment,we defrosted the granulosa cells.Using Trizol method to extract the mRNA of every example,then we reverse transcription cDNA, using Real time PCR method to detect the PFKP mRNA and LDHA mRNA gene expression of granulosa cell.ResultsThe eggs number,good oocytes rate of the patients with endometriosis were (8.2±5.6),(37.6±33.2)%, while the eggs,good oocytes rate of the controls were (11.9±5.8),(48.0±31.4)%, the eggs and good oocytes had the significant difference between these two groups (P<0.05).Obtaining eggs rate,fertilization rate,Obtaining oocytes rate were(72.4±28.3)%,(65.3±24.7)%,(98.8±5.7)%, in the endometriosis group; (73.4±22.9)%,(67.4±21.0)%,(99.4±5.4)%, in the controls.They had no differenences between the two groups (P>0.05). The pregnancy rate of the endometriosis group was 38.8%, while the The pregnancy rate of the controls was53.5% there was no differenences between the two groups (P>0.05). Adopting Western blot methods to determine the content of BMP-15 in follicular fluid of the infertile patients, the content of BMP-15 in follicular fluid of the endometriosis patients was (0.67±0.18), of the controls was (0.94±0.33), the content of BMP-15 in follicular fluid had the significant difference between these two groups(P<0.05).The content of BMP-15 in follicular fluid Using the flow cytometry analysis apoptosis rate of granulosa cell of the two groups patients, the apoptosis rate was (37.82±15.81)%, in the endometriosis groups; (8.85±5.58)%, in the controls. The apoptosis rate had the significant difference between these two groups (P<0.05). The expression of the PFKP mRNA and the LDHA mRNA in the oocytes of the endometriosis patients were (1.22±0.63) and (1.22±0.60), of the controls were(0.96±0.19)and (0.97±0.29). No statistic difference was found in the expression of PFKP mRNA and LDHA mRNA in the oocytes between endometriosis patients and controls (P>0.05).ConclusionsThe expression of BMP-15 in follicular fluid of the endometriosis patients was fewer than the controls, But the apoptosis rate of the granulosa cells of the endometriosis patients was elevated.The reducing of the BMP-15 in follicular fluid maybe affect the granulosa cell proliferation and the glycolysis,then increase the apoptosis rate of granulosa cells. All of these will affect the quality of oocytes,the growth of follicle and embryo,so pregnancy rate will decrease.
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