Changes Of Adiponectin And Its Receptor Levels In Endometrium,Ovary And Embryo And Their Effects On Conception Rates In Mice With PCOS

Part?:Testing the expression of adiponectin and its receptors in the endometrium,ovary and embryo of mice with PCOSObjective:The expression of adiponectin and its receptors in the endometrium,ovary and embryo of mice with polycystic ovarian syndrome?PCOS?were detected to observe the changes of adiponectin and its receptor levels of endometrium,ovaries and embryonic tissues in mice with PCOS during peri-implantation process of embryos,then we can investigate the effects of adiponectin on conception rates in mice with PCOS.Methods:1.A rat model of PCOS was induced by subcutaneous injection of DHEA and they were divided randomly into 4 groups:control non-pregnant group?normal untreated mice,n=10?,control pregnant group?mating after stimulating ovulation in normal mice,n=20?,PCOS non-pregnant group?only made a model of PCOS,n=10?and PCOS pregnant group?mating after stimulating ovulation in PCOS mice,n=20?.Each mouse was subcutaneously injected once a day for 4 days.2.Extracting the endometrium,ovarian tissue and pregnant mice embryos from each group on the 4th day of pregnancy.The expression of adiponectin and its receptor 1and 2 in endometrium,ovaries of each group were detected by immunohistochemical test and in embryos were detected by immunofluorescence.3.Extracting RNA from the endometrium,ovaries and embryos of each group on the4th day of pregnancy.The expression of adiponectin and its receptors mRNA were detected by Real-time PCR.4.The experimental data were expressed as mean±standard deviation and SPSS13.0was used for statistical analysis.Differences were determined by t test.A P value<0.05 was considered to be significant.Results:1.A regular estrous cycle was observed in the control group,and the PCOS group presented irregular estrous cycle.In the control group,the ovaries showed bright red,and the follicles were not obviously dilated.In the PCOS group,the appearance of endometrium was congestive.The ovaries had a pale appearance,and the volume of ovaries increased with different sizes of cystic follicles.2.The number of embryo implantation in control pregnant group was significantly more than that in PCOS pregnant group?P<0.05?.3.Immunohistochemical test showed that adiponectin?AdipoQ?,adiponectin receptor1?AdipoR1?,adiponectin receptor 2?AdipoR2?could be found in the endometrium,ovaries and embryos of mice.The PCOS group had a lower expression of AdipoQ,AdipoR1 and AdipoR2 in the endometrium and ovaries of PCOS group were significantly lower than that of the control group?P<0.05?.The expression of Adipo Q,AdipoR1 and AdipoR2 had little difference in endometrium and ovaries between PCOS non-pregnant group and pregnant group?P>0.05?.AdipoQ,AdipoR1 and AdipoR2 in embryos of PCOS pregnant group were lower than those in control pregnant group by immunofluorescence?P<0.05?.4.AdipoQ,AdipoR1 and AdipoR2 mRNA levels were measured in the endometrium,ovaries and embryos of mice.AdipoQ mRNA levels of endometrium in PCOS pregnant group were deeply lower than those in control pregnant group?P<0.05?,while AdipoR1 and AdipoR2 mRNA levels of endometrium in PCOS pregnant group were significantly higher than those in control pregnant group?P<0.05?.The levels of AdipoR1 and AdipoR2 mRNA in the ovaries of PCOS pregnant group and non-pregnant group were higher than those of control group?P<0.05?.AdipoR1 and AdipoR2 mRNA levels in embryos of PCOS pregnant group were higher than those in control pregnant group?P<0.05?.Conclusions:The expression of adiponectin and adiponectin receptors in endometrium,ovaries and embryos of mice with PCOS were lower than those in normal mice.Adiponectin may regulate the endometrium,ovarian function through the changes of AdipoR1 and AdipoR2 expression,then accordingly affect the conception rates in mice with PCOS.Part?:Preliminary observation of the effects of different concentrations of exogenous adiponectin on conception rates of mice with PCOSObjective:The expression of adiponectin and its receptors in the endometrium,ovary and embryo after subcutaneous injection of adiponectin in mice with PCOS were detected to observe the effects of different concentrations of exogenous adiponectin on conception rates of mice with PCOS.Methods:1.A rat model of PCOS was induced by subcutaneous injection of DHEA and they were divided randomly into 4 groups:Group A?control group,injecting saline,n=5?,Group B(injecting adiponectin 1?g/kg·d^-1,n=10),Group C(injecting adiponectin10?g/kg·d^-1,n=10),Group D(injecting adiponectin 100?g/kg·d^-1,n=10).2.Extracting the endometrium,ovarian tissue and pregnant mice embryos from each group on the 4th day of pregnancy.The expression of adiponectin and its receptor 1and 2 in endometrium,ovaries of each group were detected by immunohistochemical test and in embryos were detected by immunofluorescence.3.Extracting RNA from the endometrium,ovaries and embryos of each group on the4th day of pregnancy.The mRNA expressions of adiponectin and its receptors were detected by Real-time PCR.4.The experimental data were expressed as mean±standard deviation and SPSS13.0was used for statistical analysis.Differences were determined by t test.A P value<0.05 was considered to be significant.Results:1.The number of embryo implantation in group C was significantly higher than that in group A,B,D?P<0.05?.2.AdipoQ?AdipoR1 and AdipoR2 in the endometrium of group B,group C and group D were higher than those of group A?P<0.05?.AdipoQ?AdipoR1 and AdipoR2 in the endometrium and ovaries of group C were higher than those of other groups?P<0.05?.There were no significant differences on the expressions of AdipoQ,AdipoR1 and AdipoR2 in embryos amang those groups?P>0.05?.3.AdipoQ,AdipoR1 and AdipoR2 mRNA levels in the endometrium,ovaries and embryos of group A were lower than those of other groups?P<0.05?.The levels of AdipoQ,AdipoR1 and AdipoR2 mRNA of group C were higher than those of group B and group D?P<0.05?.AdipoQ,AdipoR1 and AdipoR2 mRNA levels in embryos of group D were lower than those of control group?P<0.05?.Conclusions:Moderate concentration of exogenous adiponectin could increase the expressions of adiponectin and its receptors in the endometrium,ovary and embryo to enhance the conception rates of mice with PCOS.Higher concentration of exogenous adiponectin could decrease the expressions of adiponectin and its receptors in the endometrium,ovary and embryo and reduce the conception rates of mice with PCOS.