Study On The Regulation And Related Mechanism Of DHEA On Oxidative Stress Endometrial Cell Receptivity In Mice

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The effect of Dehydroepiandrosterone(DHEA)supplementation on women with diminished ovarian reserve(DOR)in IVF cycle: Evidence from a meta-analysisObjective In order to evaluate that whether DHEA supplemtntation could improve the ovarian response and pregnancy outcome in diminished ovarian reserve(DOR).Methods PubMed?EMbase?Cochrane Library were searched before 31.8.2015,the researches that compared pre-treated with DHEA or not to DOR patient in IVF cycle,included RCTs and observation studies,were selected for our meta-analysis.Outcome measures were the number of retrieved oocytes,cancellation rate of IVF cycles,clinical pregnancy rate and miscarriage rate.We adopted Revman 5.0 software to pool the data from the eligible studies.The data was analysed by Rev Man 5.0.Results Nine studies were eligibled for analysis,four RCTs,four observation studies and one prospective study non RCT.Funned plot showed unsymmetry,the publication of potential deviation could not be excluded.A total of 1208 patients,540 in DHEA groups,668 in control groups.Combinded the results of analysis showed that DOR patient pre-treated with DHEA,the clinical pregnancy rate in DHEA groups was significant higher than control groups(OR=1.47,95%CI: 1.09-1.99),the heterogeneity between each study was small(I2 20%).However,the numbers of oocytes retrieved,the cancellation rate of IVF cycles and the miscarriage rate were no difference between the two groups(WMD=-0.69,95%CI:-2.18-0.81;OR=0.74,95%CI: 0.51-1.08;OR=0.34,95%CI: 0.10-1.24).Combinded the results of oocytes retrieved analysis showed that there was significant heterogeneity between each study(I2 97%),but little in he cancellation rate of IVF cycles and the miscarriage rate.It is worth noting,when data were restricted to RCTs,there was a non-significant difference in the clinical pregnancy rate(OR=1.08,95%CI: 0.67-1.73),there was no heterogeneity between each study(I2 0%).Conclusion We concluded that DHEA supplementation in DOR patients might improve the pregnancy outcomes.To further confirm this effect,more randomized controlled trials with large sample sizes are needed.Part ? The effect of oxidative stress on mouse decidual endometrial stromal cellObjective The purpose of the study was to establish the oxidative stress state of decidual mouse endometrial stromal cells(ESCs),determine the proper action concentration of H2O2,as a oxidative stress revulsant,and to provide the foundation for further research the relationships between DHEA,oxidative stress and the receptivity of endometrial.Methods Primary ESCs were induced decidualization by 10 n M E2?1?M P4 for 72 h in vitro after successfully isolated and cultured from mice.Different concentration of H2O2 was added.The primary ESCs was identified by immunohistochemistry,the expressing of d PRP m RNA was analyzed by PT-PCR,the growth and proliferation of decidual ESCs treated with H2O2 was analyzed by CCK-8 and the level of ROS was analyzed by flow cytometry.The data was analysed by SPSS 20.0.Result Primary mouse ESCs were successfully isolated and cultured,it was spidle and polygon,radial alignment under microscope.Immunofluorescence analysis showed that expression of Vimentin(+),Cytokeratin(-).The purity of primary mouse ESCs was(96.3±0.49)%.The expression of d PRP m RNA was distinctly higher when primary mice ESCs treated with 10 n M E2?1?M P4,the difference was statistically significant(2.0437±0.0110 vs 1.0001±0.0000;t=-164.202,P<0.01).The ability of growth and proliferation of decidual primary ESCs was distinctly repressed when the concentration of H2O2 was greater or equal to 150 ?mol/L.The maximum inhibitory was about 70.6% at the concentration of 300?mol/L.Concentrations of intracellular ROS are significantly increased when decidual primary endometrial stromal cells treated with 50 ?mol/L?100 ?mol/L H2O2,the difference was statistically significant(27.77±4.20 vs 17.47±0.61;43.57±6.58 vs 17.47±0.61,P<0.01).Conclusion 100 ?mol/L was the proper concentration of H2O2 for further research that can significantly produce intracellar ROS and not affect the ability of growth and proliferation of decidual primary endometrial stromal cells.Part ? Studies of the effect and mechanism of DHEA on mouse decidual ESCs receptivityObjective To investigate the effect of dehydroepiandrosterone(DHEA)on mouse decidual endometrial stromal cells(ESCs)and to explore mechanisms regulating endometrial receptivity.Methods Primary ESCs were treated with different concentrations of DHEA and flutamide(FLU),a specific androgen receptor(AR)antagonist during inducing decidualization by 10 n M E2?1?M P4 for 72 h in vitro after being successfully isolated and cultured from mice.After decidualization,100 ?mol/L H2O2 was added to the ESCs.Flow cytometry was used to measure intracellular reactive oxygen species(ROS).Real time-PCR was used to determine m RNA expression of decidual PRL-related protein(d PRP),AR,and Homeobox A10(HOXA10).Protein levels of AR and HOXA10 were measured by western blot.Results Primary mouse ESCs were successfully isolated and induced decidualization.The ability of growth and proliferation of decidual primary ESCs were significantly inhibited at the conditions of DHEA ?1×10-6 M with a trend of concentration dependent,the maximum inhibitory ratio was about 84% at the concentration of 1x10-3 M,there was a significantly statistical difference(P<0.01).The ability of proliferation of was not affected when consentrations of DHEA are from 1×10-9 to 1×10-5M,the intracellular ROS of Decidual the mice induced by 100 ?mol/L H2O2 was significantly reduced in a dose-dependent manner when treated with 10 n M DHEA or 100 n M DHEA(34.70 ± 5.57 RIF;25.50 ± 6.42 RIF),There is a significant statistical difference(P < 0.01).The expression of d PRP was minimally affected by DHEA at concentrations of 1 to 100 n M.However,expressions of HOXA10 m RNA(2.0380 ± 0.2215 vs 1.0029 ± 0.0041)and protein(0.0545 ± 0.0085 vs 0.0152 ± 0.0037)were significantly increased by 100 n M DHEA,there was significantly statistical difference(P<0.01).Importantly,the increase of HOXA10 m RNA and protein expression induced by DHEA can be inhibited by treatment with FLU,but,neither DHEA nor FLU influenced the expression of AR m RNA or protein.Conclusion Low concentration of DHEA improves the antioxidant capacity of decidual ESCs.DHEA treatment may also improve endometrium receptivity via AR.