Effect Of Qigong Pill On The Expression Of PDCD4 And PCNA In Ovarian Granulosa Cells In PCOS Patients With Phlegm-dampness Syndrome

Objective:To observe the effect of Qigong Pill on the expression of programmed cell death 4（PDCD4）and proliferating cell nuclear antigen（PCNA）in ovarian granulosa cells（GCs）with polycystie ovarian syndrome（PCOS）.From GCs’proliferation and apoptosis of phlegm-dampness PCOS,to explore the mechanism of Qigong Pill in improving the ovulation function and the quality of oocytes.Methods:A total of 66 patients with phlegm-dampness PCOS undergoing in vitro fertilization and embryo transfer（IVF-ET）were selected and divided into observation group（Qigong Pill+western medicine）and control group（western medicine）according to the random number table method,33 patients in each group,all groups were treated with the inhibitor protocol for controlled ovarian hyperstimulation.The observation group taked Qigong Pill one month before the IVF cycle to the day of egg collection,while the control group did not take traditional Chinese medicine.Recording the general information,serum basal endocrine level and serum anti-mullerian hormone（AMH）,counting body mass index（BMI）,and scoring the phlegm-dampness syndrome before and after treatment.The dosage and the number of days of gonadotropins（Gn）,luteinizing hormone（LH）,estradiol（E₂）and progesterone（P）levels on the day of HCG injection,the number of oocytes obtained,2PN fertilization rate,high-quality embryo rate were calculated and compared between the two groups.At the same time,the ovarian GCs were separated and extracted on the day of HCG injection,using quantitative-real time polymerase chain reaction（Q-PCR）and western blot（WB）techniques to detect the expressions of PCNA and PDCD4 in the two groups of GCs.Results:1.Phlegm-dampness syndrome score:the scores of phlegm-dampness syndrome in the observation group after treatment were significantly lower than those before treatment,and the difference has statistics significance（p<0.01）;there was no significant difference in the scores of phlegm-dampness syndrome before and after treatment in the control group（p>0.05）;after treatment,a statistically significant difference between the observation group and the control group in the score of phlegm-dampness syndrome（p<0.01）.2.Gn days and Gn dosage:the difference between the observation group and the control group was not statistically significant（p>0.05）.3.Serum LH,E₂and P levels on the day of HCG injection:the E₂ level of the observation group were higher than that of the control group,and the difference was statistically significant（p<0.01）,but the LH level and the P level was not statistically significant（p>0.05）.4.The number of oocytes obtained:there was no significant difference in the number of oocytes obtained between the two groups（p>0.05）.5.The 2PN fertilization rate and high-quality embryo rate:the 2PN fertilization rate and high-quality embryo rate of the observation group were significantly higher than those of the control group,and the difference was statistically significant（p<0.01）.6.The relative expression levels of PDCD4 and PCNA genes and proteins in the ovarian GCs:compared with the control group,the expression of PDCD4 and PCNA was down-regulated in the observation group,and the expression of PCNA was up-regulated.These differences between the two groups were statistically significant（p<0.05）.Conclusion:In IVF-ET,Qigong Pill can increase the 2PN fertilization rate and high-quality embryo rate of PCOS patients with phlegm-dampness syndrome.It may be through down-regulating the expression of PDCD4 and enhancing the expression of PCNA in ovarian GCs to intervene the development of follicles,regulate hormone levels,inhibit the apoptosis and promote the growth of GCs cells in PCOS patients with phlegm-dampness syndrome,thus improves the quality of oocytes and embryonic development potential.