Clinical And Mechanism Study On Guiyuan Decoction In The Treatment Of Decreased Ovarian Reserve Functio

Part 1:Clinical studyObjective:To observe the effects and safety of Guiyuan decoction in the treatment of decreased ovarian reserves.Methods: Clinical research used randomized control as methods.158 patients with DOR received by Longhua Hospital affiliated to Shanghai University of Traditional Chinese Medicine,randomly into Guiyuan decoction group(53 cases),Complex Packing Estradiol Tablets/Estradiol and Dydrogesterone Tablets(Femoston)group(52 cases),combination treatment Group(53 cases).Used corresponding medicine for 3 months.Before and after treatment,Observed serum hormones,anti-Mullerian Hormone(AMH),antral follicle count(AFC),ovarian hemodynamic indicators,endometrial thickness,Chinese medicine syndromes scale,menstrual quantitative score,anxiety self-evaluation scale(SAS),depression self-evaluation scale(SDS),and security indicators.Results:1.After treatment,three groups of serum exgrad(E2)were higher(P <0.01),serum follicle stimulating hormone(FSH),ratio of serum follicle stimulating hormone to lurosulent hormone(FSH / LH)were decreased(P <0.01).The E2 level had no difference.The combination of pharmaceutical group and Femoston group were more obvious(P <0.05).2.Three sets of AMH and AFCs increased after treatment(P <0.01).There was no difference between AFC levels.The combination medicine group compared with Chinese medicine groups and Femoston group had better efficacy in raising AMH(P<0.05).3.After treatment,three groups of Chinese medical test scores,menstrual quantitative scores were lower(P <0.01).Chinese medicine group and combination medicine group,had better score compared wiyth Femoston group(P <0.05).The scores of Chinese medicine syndrome were 50.94%(Chinese medicine group),17.31%(Femoston group),45.28%(combination medicine group)and the difference is significantly different(P ’<0.0125).4.Three groups of ovarian hemodynamic indicators: the resistance index(RI),the pulsatility Index(PI)were lower,and the peak systolic velocity(PSV)were higher,endometrial thickness were increased(P <0.01).There were no difference between RI and PI.Chinese medicine group raised PSV and endometrial thickness more than Femoston group(P <0.05).5.Guiyuan decoction group and combination of pharmaceutical group had lower SAS,SDS score after treating(P <0.01).There was no significant improvement of SAS,SDS score with Femoston(P> 0.05).6.The total efficiency of the Chinese medicine syndrom were: 83.02%(Guiyuan decoction group),59.62%(Femoston group),86.79%(combination medicine group),Combination medicine group and Chinese medicine group were more effective(P’<0.0125).7.Safety indexes:there were no abnormalities in blood routine,urine routine,liver and kidney function in the three groups.During the treatment,one case of dyspepsia occurred in Guiyuanjian group and combination medicine group(bloating and irregular stool dryness and thinning,respectively);One patient had migraine and one patient had itchy skin in Femoston group.Symptoms all decreased after symptomatic treatment.Conclusion:1.Guiyuan decoction can improve the ovarian reserve function of DOR,and have safety.2.Compared with Femoston group,There is obvious advantage of Chinese medicine in adjusting the ovarian blood flow,increasing the thickness of the endometrium,reducing clinical symptoms,reducing anxiety depression.3.Combination of pharmaceutical group can significantly increase the serum AMH and reduce clinical-related symptoms.Part 2:Experiment studyObjective:To investigate the effects and mechanism of Guiyuan decoction on mice with decreased ovarian reserve.Methods:60 female ICR mice were randomly divided into control group,model group,estradiol valerate group(0.15 mg/kg)and low,medium,high dose groups of Guiyuan decoction(0.75、1.49、2.98 g/kg),with 7 mice in each group.Except the control group,cisplatin was injected intraperitoneally to establish model of decreased ovarian reserve.After modeling,mice in estradiol valerate group and Guiyuanjian decoction groups were given 0.3 m L of corresponding drugs,mice in model group and control group were given the same volume of normal saline.Intragastric administration were taken once a day for 4 weeks.After the last Administration,enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of anti-mü llerian hormone(AMH)and follicle-stimulating hormone(FSH)in mouse serum,histopathological morphology of ovarian were observed by hematoxylin-eosin(HE)staining,protein distribution of AMHRII and Smad4 in ovarian tissue were observed by immunohistochemistry;protein quantity of AMHRII and Smad4 were detected by Western blot.The mitochondrial structure of ovarian granulosa cells was observed by transmission electron microscope.Result:1.Compared with control group,level of serum AMH,expression of AMHRII and Smad4 protein in ovarian tissue in model group,were significantly decreased,while the FSH level in serum was significantly increased(P<0.05).Follicles were crumpled and lost nucleus,ovarian interstitial were fibrosis,luteum were loose,AMHRII and Smad4 protein in ovarian tissue were mainly distributed in the follicle membrane and ovarian interstitial.2.Compared with the model group,expression of AMHRII and Smad4 protein in ovarian tissue was ascended,serum AMH were rising,FSH were descended in estradiol valerate group and Guiyuan decoction groups.Ovarian tissue morphology of each administration group were improved,mature follicles grow,AMHRII and Smad4 were mainly distributed in the granular cell membrane and follicular nucleus.3.DOR model mice caused by Cisplatin had destroyed mitochondrial of ovarian particulate cells.The shape and function of mitochondrial were damaged.In each treatment group,various amounts of mitochondrial showed up,but the morphology did not improve significantly.Conclusion:Guiyuan Decoction has a certain effect on improving ovarian function in DOR model mice.The mechanism may be related of promoting serum AMH,increasing number of AMHRII and Smad4 receptors,reducing FSH levels...