The Effect Of Total Flavonoids Of Dodd Vder Seed On Rat Model Of Polycystic Ovary Syndrome

Objective:To observe the effect of total flavonoids of dodder seed on rat model of polycystic ovary syndrome?PCOS?induced by dehydroepiandrosterone combined with human chorionic gonadotropin and Letrozole,and to investigate the therapeutic effect and characteristics of total flavonoids of dodder seed on PCOS.Methods:Using dehydroepiandrosterone combined with human chorionic gonadotropin to establish the rat model of PCOS.70 21-day-old female SD rats were fed to the age of 23 days,ten rats of which were randomly selected as the blank control group,and the rest of which were used as model groups.On every morning,hypodermic injecting DHEA6mg/?100g.D?+0.2mL injectable oil agent on the back of the neck,and in the afternoon,hypodermic injecting 1.5 IU/d HCG+0.4mL aquae pro injectione.The moulding lasted 21 days.Rats in blank control group were injected with equal volume of solvent injection.From the 16th day of modeling,observing the vaginal smears of rats for 1 cycle.The criteria for the success of the model were as follows:Vaginal epithelium continues to keratinize.Selecting fifty rats from the successful model and randomly divided into 5 groups:model group,Diane-35 group(0.3392mg·kg^-1),total flavonoids of dodder seed large,medium and small dose groups(0.2g·kg^-1,0.1g·kg^-1,0.05g·kg^-1),10 in each group.Except blank group and model group,the other groups are given corresponding drugs for 3 weeks.2 hours after the last administration,the rats are weighed and anesthetized.then to determine the level of E2,INS,LH,FSH,P and PRL in serum.To remove the uterus and ovary and calculate its viscera index.After removal of ovary,uterus and pancreas,observe HE staining and record the morphological changes of each organ under light microscope.In addition,by immunohistochemistry,to observate the mean optical density of ovary AR,BCL-2,Bax,TNF-?,IGF-l,and hypothalamus GnRH,pituitary GnRH-R.Using Letrozole to establish the rat model of PCOS.Seventy 6-week-old female SD rats were fed to the age of 23 days,ten rats of which were randomly selected as the blank control group,and the rest of which were used as model groups.Model rats were given letrozole1mg/?kg.d?daily,The blank group was infused with the same amount of solvent for21d.From the 16th day of modeling,observing the vaginal smears of rats for 1 cycle.The criteria for the success of the model were as follows:Vaginal epithelium continues to keratinize.Selecting fifty rats from the successful model and randomly divided into 5 groups:model group,Diane-35 group(0.3392mg.kg^-1)and total flavonoids of dodder seed large,medium and small dose groups(0.2g·kg^-1,0.1g·kg^-1,0.05g·kg^-1),10 in each group.Except blank group and model group,the other groups are given corresponding drugs for 3 weeks.2 hours after the last administration,the rats are weighed and anesthetized.then to determine the level of T,E2,LH,FSH,P,PRL,INS,GnRH,TNF-?,HDL-C and LDL.To remove the uterus and ovary and calculate its viscera index.After removal of Right ovarian tissue,adrenal tissue,pancreatic tissue,observe HE staining and record the morphological changes of each organ under light microscope.In addition,by immunohistochemistry,to observate the mean optical density of AR in the hypothalamus,pituitary,and left ovary.Results:The rat model of PCOS induced by dehydroepiandrosterone combined with human chorionic gonadotropin was successfully established.The model group was compared with the blank group,in the model group,the indexes of ovary and uterus in the model group were significantly higher?P<0.01?,The serum levels of serum T,E2,LH,LH/FSH,PRL and INS were significantly higher?P<0.01?,P level decreased significantly?P<0.01?,Serum FSH level decreased obviously?P<0.05?,The mean optical density of GnRH in hypothalamus was significantly increased?P<0.01?,the mean optical density of GnRH-R in pituitary was significantly decreased?P<0.01?,and the mean optical density of AR,Bax,TNF-?and IGF-l in ovary was significantly increased?P<0.01?,but in ovary BCL-2 decreased significantly?P<0.01?.Compared with the model group,larger,medium,small dose group of total flavonoids of dodder seed and Diane-35group could significantly reduce the ovarian and uterine index of PCOS in rats?P<0.01?,and significantly decreased the level of serum T,E2,LH,LH/FSH,PRL and INS Significant increase in serum P level?P<0.01?.The larger,medium dose group of total flavonoids of dodder seed can obviously increase serum FSH level?P<0.05?.The larger,medium,small dose group of total flavonoids of dodder seed can significantly decrease the mean optical density of GnRH in hypothalamus and AR,Bax and TNF-?in ovary in rats with PCOS.The larger,medium dose group of total flavonoids of dodder seed can obviously increase GnRH-R in pituitary and BCL-2in ovary?P<0.05?and significantly decrease mean optical density of IGF-1 in ovary?P<0.01?.The rat model of PCOS induced by Letrozole was successfully established.The model group was compared with the blank group,the ovarian index was significantly increased?P<0.01?,Uterine index decreased significantly?P<0.01?,the level of T,LH,LH/FSH ratio,LDL,PRL,INS,GnRH and TNF-?in serum was significantly increased?P<0.01?.The serum level of P,E2 and HDL-C of rats in the model group was significantly decreased.The level of FSH in serum was obviously decreased?P<0.05?.Compared with the model group,in larger,medium,small dose group of total flavonoids of dodder seed,the ovarian index of rats decreased significantly?P<0.01?,Uterine index increased significantly?P<0.01?,the serum level of LH,LH/FSH ratio and T were significantly decreased in rats?P<0.01?,the serum level of PRL,LDL and GnRH significantly reduced?P<0.01?,the serum level of P and HDL-C Significantly increased?P<0.01?.in the larger,medium dose group of total flavonoids of dodder seed,the serum level of INS and TNF-?significantly reduced?P<0.01?,the serum level of E2 increased significantly?P<0.01?and FSH level increased obviously?P<0.05?;the larger,medium,small dose group of total flavonoids of dodder seed can significantly reduce average optical density of AR in hypothalamus,pituitary and ovary of PCOS rats?P<0.01?.Conclusion:It is suggested that the total flavonoids of dodder seed can improve the effect of dehydroepiandrosterone combined with human chorionic gonadotropin and letrozole on the model of PCOS in rats,and improve the PCOS caused by high androgen level and hyperfunction of hypothalamus-pituitary-ovary axis.the total flavonoids of dodder seed also can effectively relieve infertility caused by PCOS.