Study On The Mechanism Of Cangfu Daotan Decoction In Patients With Phlegm-dampness Polycystic Ovary Syndrome Based On HSP70/NF-κB/Caspase-3 Signaling Pathway

Objective: Look for the differences in the expression of the factors in HSP70/NF-κB/Caspase-3 pathway in ovarian granulosa cells(GCs)of patients with phlegm-dampness type of Polycystic Ovary Syndrome(PCOS),and to explore the function of granular cell apoptosis in the pathogenesis of phlegm-dampness type of PCOS,to clarify the molecular mechanism of Cangfu Daotan Decoction granule in the treatment of phlegmdampness type of PCOS.Method: A total of 66 patients with phlegm-dampness type of PCOS who planned to receive assisted reproductive technology for pregnancy were selected and randomly divided into the treatment arm(Cangfu Daotan Decoction granules + IVF-ET)and the control arm(placebo granules + IVFET),with 33 patients in each group.33 normal patients who received IVFET only for male factors were selected as blank control arm(placebo granule + IVF-ET).Antagonist regimen was adopted in all three groups.The phlegm-dampness syndrome scores of PCOS two groups before and after treatment were compared,and the levels of estradiol(E2),luteinizing hormone(LH),progesterone(P),total amount and days of gonadotropin(Gn)on HCG day,body mass index(BMI),number of follicle obtained,2PN fertilization rate and fine embryo rate of the three groups were compared.The expression levels of heat shock proteins 70(HSP70),nuclear transcription factor-κB(NF-κB),NF-kappa-B inhibitor alpha(IкBα)and Caspase-3 in GCs were determined and statistically compared.Results:1.Phlegm-dampness syndrome: There was statistical significance in the phlegm-dampness syndrome score between the treatment arm and the control arm after treatment(P<0.05);The phlegm-dampness syndrome score after treatment in the treatment arm was significantly lower than that before treatment(P < 0.01);There was no statistical significance in the phlegm-dampness syndrome score in the control arm before and after treatment(P > 0.05).2.BMI: BMI of treatment arm and control arm was significantly higher than that of blank control arm before treatment(P<0.01),but there was no significant difference in BMI between the treatment arm and control arm before treatment(P>0.05);The BMI of the treatment arm was significantly decreased after treatment(P<0.01),while the BMI of the control arm and the blank control arm was not significantly changed before and after treatment(P>0.05).3.Sex hormone levels among the three groups on trigger day: there were no significant differences in LH and P levels among the three groups on trigger day(P>0.05);E2 level of the treatment arm was higher than that of the blank control arm(P < 0.05),but there was no significant difference between the control arm and the treatment arm,and between the control arm and the blank control arm(P>0.05).4.Dosage and days of Gn in the three groups: The total amount and days of Gn were not significantly different among the three groups(P>0.05).5.Comparison of IVF-ET related data: The number of follicle obtained in the treatment arm and the control arm was higher than that in the blank control arm(P< 0.05),there was no significant difference between the treatment arm and the control arm(P>0.05);2PN fertilization rate and high-quality embryo rate of the treatment arm and blank control arm were increased compared with the control arm(P<0.05),and no significant differences were found between the treatment arm and the control arm(P>0.05).6.Comparison of the m RNA and protein contents of HSP70,NF-κB,Caspase-3and IкBα in GCs of three groups: The m RNA and protein contents of HSP70,NF-κB,Caspase-3 and IкBα in control arm were significantly increased compared with the other two groups(P < 0.05);The m RNA and protein contents of HSP70,NF-κB,Caspase-3 and Iк Bα in treatment arm were higher than those in blank control arm(P<0.05).Conclusions: 1.Overexpression of HSP70,NF-κB,IкBα and Caspase-3increased the apoptosis rate of GCs in PCOS patients,may cause the occurrence of PCOS.2.Cangfu Daotan Decoction granules can improve the phlegm-dampness symptoms of PCOS patients,and optimize the outcome of IVF-ET.3.The therapeutic effect of Cangfu Daotan Decoction granules on PCOS patients may be achieved by reducing the contents of HSP70,NF-κB,IкBα,Caspase-3 in ovarian GCs to reduce the apoptosis of GCs.