| Objectives:To obserce the effect of reinforcing kidney Chinese Medicine on Mfn2 and mitochondrial membrane potential of granular cell in aged IVF patients with kidney qi and yin deficiency,and to explore the molecular mechanism of Chinese Medicine on clinical outcomes from perspectives of energy metabolism.Methods:(?)Clinical study:76 patients underwent IVF-ET with kidney qi and yin deficiency were randomly and equally divided into treatment group(Erzhi Tiangui Granule combined with IVF)and placebo group(placebo combined with IVF),another 43 cases caused by male factors were chosen as normal control group(placebo combined with IVF either).After the treatment of Gonadotropin-releasing Hormone Antagonist Protocol(GnRH-ant)until the human chorionic gonadotropin(hCG)injection day,syndrome score of kidney qi and yin deficiency,the number of oocytes,the ratios of cleavage?high-quality embryos and clinical pregnancy,time parameter and cleavage patterns were observed.(?)Mechanistic study:On oocyte retrieval day,granulosa cells of each groups were collected.Expression of Mfn2 in mRNA and protein level in granulosa cells were measured by real-time reverse transcription-polymerase chain reaction(RT-PCR)and Western-blot.Changes of mitochondrial membrane potential(??m)were observed by JC-1 staining method.Results:(?)1.Syndrome score of kidney qi and yin deficiency were significantly improved in the treatment group,and compared with the placebo group,changes of syndrome score before and after treatment were statistically significant(P<0.01).2.Compared with the placebo groups,the ratios of 2PN cleavage?high-quality embryos and clinical pregnancy in IVF were slightly increased in the treatment group(P>0.05),while in the normal control group,the differences were statistically significant(P<0.05).3.Compared with the placebo group,the ratios of 2PN cleavage and high-quality embryos in ICSI in the other two groups were statistically significant(P<0.05),while there was no significant difference between the treatment groups and normal control group(P>0.05).4.There were no significant differences among three groups in t2 and s2 period(P>0.05).Compared with the other two groups in t3,t4 and cc2 period,the development in normal control group were significantly faster(P<0.05).Compared with the placebo group in t5 period,development in the other two groups were statistically faster(P<0.05),while in normal control group,it is much faster than the treatment group,indicating a significant difference(P<0.05).5.Cleavage patterns in the normal control group were significantly higher than in placebo group(P<0.05),but in the treatment group,it is slightly higher with no statistically difference compared with the placebo group(P>0.05).(?)1.Compared with the placebo group,expression of Mfn2mRNA and protein were both significantly higher in the other two groups(P<0.05),while there was no significant difference between the treatment groups and normal control group(P>0.05).2.Compared with the placebo group,the fluorescence intensity ratio of ??m were significantly higher in the other two groups(P<0.01),while it is also statistically higher in the normal control group than in the treatment group(P<0.01).Conclusion:1.In IVF-ET treatment cycle,Erzhi Tiangui Granule for tonifying kidney could improve symptoms of kidney qi and yin deficiency,increase the number of oocytes,improve the ratios of 2PN cleavage?high-quality embryos and clinical pregnancy,slow down early embryonic development block in aged women and decrease the ratio of abnormal cleavage.2.Reinforcing Kidney Chinese Medicine may improve the outcomes of aged patients underwent IVF with kidney qi and yin deficiency by increasing expression of ??m and Mfn2 in granulosa cells,promoting the process of mitochondrial fusion,inhibiting the decline of ATP level and slowing down the decrease of ??m in order to promote the development of oocytes and improve and developmental potential of early embryos. |
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