Study On The Mechanism Of Sequential Therapy Of Nourishing-Yin And Supplementing-Yang Intervention In Follicular Development With PCOS

Objective:Through sequential therapy of nourishing Yin and supplementing Yang of traditional Chinese medicine intervention to study the mechanism of the therapy controlling secretion regulation function and expression of cytokine genes in ovarian granulosa cells in PCOS rats.Furthermore, monitor stem cell factor(SCF) level in serum,follicular fluid by ovulation induction cycle or in vitro fertilization and embryo transfer (IVF-ET) cycle on infertile patients with PCOS,besides its relationship with the influence of dominant follicle and oocyte quality.Methods:1. Experiment Study:According to the experimental requirements establish PCOS rats model,then, isolate and culture ovarian granulosa cells in different periods.Prepare three different medicated serum namely blank serum, nourishing Yin serum, supplementing Yang serum, to observe their effects on PCOS rats’ovarian granulosa cell secretion function of E2, P,and the influence of stem cell factor, androgen receptor, steroidogenic acute regulatory protein gene expression.2. Clinical research:30cases of classic PCOS,30cases PCOS without hyperandrogenism,30patients without PCOS were selected to analysis correlation of SCF and PCOS pathogenesis in basic state. Another62cases PCOS patients are randomly divided into the treatment group (sequential therapy of nourishing Yin and supplementing Yang of traditional Chinese medicine/clomiphene citrate/Gonal-F,30cycles) and control group (clomiphene citrate/,Gonal-F,32cycles).Two groups were observed with treatment days, endometrial thickness, the dominant follicles number,serum E2, LH, SCF levels in the HCG injection day and compared ovulation rate, pregnancy rate,ovarian hyperstimulation syndrome (OHSS) and luteinized unruptured follicle syndrome (LUFS) etcetera.40cases of PCOS patients of IVF-ET were randomly divided into into2groups, experimental group (sequential therapy of nourishing Yin and supplementing Yang of traditional Chinese medicine with western medicine group) and control group (Western medicine group), each group of20cases. Observation serum E2, LH, P levels in HCG injection day,SCF levels in follicular fluid and the difference of retrieved oocyte, fertilization rate, cleavage rate, high quality embryo rate and pregnancy rate.Results:1. PCOS rat ovarian anatomy and HE staining light microscope ovarian are obviously different.2. In follicular phase, nourishing Yin Decoction10%,20%groups can improve PCOS rats’E2, SCF content in granulosa cell,similarly in luteal phase, supplementing Yang Decoction20%group displayed the strongest function of stimulating granulosa cells to secret P,but no enhancement when concentration increasing. No statistical significance among the groups of SCF level.3. There were significantly reduced the granulosa cells secretion of E2and P in model group.Both nourishing Yin Decoction10%,20%groups and supplementing Yang Decoction20%group can promote PCOS rats’ ovarian granulosa cells to secret E2, P.And that nourishing Yin Decoction concentration has a certain dose-effect relationship with granulosa cell secretion level of E2, P.4. Expression of AR mRNA in model group was significantly increased, but SCF, StAR mRNA expression decreased obviously.while,nourishing Yin Decoction10%,20%groups and supplementing Yang Decoction20%group can effectively reduce the granulosa cell AR mRNA expression, increase SCF, StAR mRNA expression.5. The without hyperandrogenism PCOS group AFC was significantly more than the classical PCOS group, normal group,and had a significantly elevated SCF levels,show a positive correlation with AFC. But,its T level,besides the normal group,was significantly lower than the classical PCOS group.6. The treatment group’s cure rate was36.7%and total efficiency of86.7%,meanwhile,control group, the cure rate was12.5%, the total efficiency of87.5%, two group had significant difference in the cure rate, endometrial thickness,above14mm and18mm follicles number in HCG injection day. Treatment group’s serum E2, SCF level in HCG injection day were significantly higher than those in control group. The test group was higher than that of the control group pregnancy.7. In IVF-ET cycles, the follicular fluid levels of SCF in retrieved oocyte day, fertilization rate, high quality embryo rate was significantly higher in treatment group han that of control group. In either group,pregnant patients’follicular fluid SCF levels were significantly higher than those not pregnant in retrieved oocyte day.Conclusion1. SCF relates to the pathogenesis of PCOS follicles developmental disorders.2. Sequential therapy of nourishing Yin and supplementing Yang of traditional Chinese medicine intervention can significantly improve the PCOS rats’ ovarian granulosa cells secretion function, down-regulate AR gene expression, up-regulate SCF, StAR gene expression.3. Sequential therapy of nourishing Yin and supplementing Yang of traditional Chinese medicine with western medicine to induce ovulation can increase the endometrial thickness, serum E2, SCF levels in HCG injection day, improve the pregnancy rate.4. IVF-ET patients with PCOS,intervent by sequential therapy of nourishing Yin and supplementing Yang of traditional Chinese medicine, can improve the ovarian microenvironment, improve the SCF level in follicular fluid, so as to improve the fertilization rate, high quality embryo rate, pregnancy rate.