Effects Of Pre-Transplant Acupuncture On Methylation Of Imprinted Genes For Ivf In Mouse Oocyte With Poor Ovarian Response

Objectives:With the growing fertility needs of poor ovarian response patients,acupuncture was confirmed that has definite effects on promoting follicular development and improving oocyte quality,but no breakthrough has been made in its deep mechanism.This study evaluated the effect of pre-transplant acupuncture intervention on ovarian reserve function and methylation of imprinted genes in mice with poor ovarian response,which aims to explore the mechanism of acupuncture to improve in vitro fertilization(IVF)effect.Methods:Female adult C57 BL/6N mice were randomly divided into 4 groups: ovarian stimulation group(group CP),acupuncture and ovarian stimulation group(group CA),modeling and ovarian stimulation group(group CM),and modeling and acupuncture and ovarian stimulation group(group MA).After adaptive feeding,each group was given the corresponding interventions of modeling with tripterygium wilfordii and acupuncture,PMSG and h CG were used for ovarian stimulation,and oocytes were collected for counting and single-cell methylation sequencing.The number of oocytes and the proportion of the second meiotic metaphase oocytes were observed.At last,the methylation levels of imprinted genes such as Mest/Peg1,Igf2 r,Peg3 and Snrpn were analyzed.Results:After promoting the eggs,we count the total number of eggs and MII egg number in the microscopic observation,and it was found that:(1)Compared with group CP,group CA obtained by MII egg number and mature oocytes proportion of eggs that there was no significant difference(P > 0.05),but the total number of oocytes was significantly lower than group CP(P < 0.05).(2)The total number of oocytes and the number of MII oocytes in group CM and group MA were significantly lower than those in group CP(P < 0.05),but the proportion of mature oocytes in group MA was not significantly different from that in group CP(P > 0.05),while group CM was significantly lower than it(P < 0.05).(3)Compared with group MA,the total number of oocytes,the number of MII oocytes,and the proportion of mature oocytes in group CM were all lower,and the differences were statistically significant(P < 0.05).(4)Compared with the MA group,group CA had a higher total number of oocytes,the number of MII oocytes,and the proportion of mature oocytes,and the difference was statistically significant(P < 0.05).In addition,the distribution of the imprinted genes Mest/Peg1,Igf2 r,Peg3 and Snrpn in the DMR fragments was statistically analyzed,and it was found that:(1)compared with group CP,the methylation levels of DMR in Mest/Peg1,Igf2 r and Snrpn in group CA were all significantly decreased.(2)Compared with group CP,the methylation level of DMR in Mest/Peg1 fragment was significantly decreased in group CM.(3)Compared with group CA,the DMR methylation level of group MA in Peg3 fragments decreased significantly.(4)Compared with group CP,there was a significant decrease in DMR methylation levels in Mest/Peg1,Igf2 r and Peg3 fragments in group MA,and it was found that the methylation levels in group CP were significantly lower than those in group MA.(5)No DMR was found between group CM and group MA.Conclusions: 1.Acupuncture intervention before transplantation has a significant effect on poor ovarian response mice,and its main mechanism is to enhance ovarian reserve function and improve oocyte quality.In addition,acupuncture intervention also reduced the ovarian reserve function of normal mice,which may be related to the stress effect.2.The mechanism of acupuncture therapeutic effect in poor ovarian response mice before transplantation was independent of methylation modification of imprinted genes such as Mest/Peg1,Igf2 r,Peg3 and Snrpn,but the stress effect of acupuncture on mice may be promoted by inhibiting methylation modification of Mest/Peg1,Igf2 r and Snrpn.