Effects Of Erzhi Tiangui Granule On Diminished Ovarian Reserve Based On MicroRNA Expression Profile

Objective:To explore the difference of microRNA expression between the patients with DOR and the women with NOR,and to apply the intervention of Erzhi Tiangui granule,to explore the influence of kidney tonifying herbs on the decrease of ovarian reserve function,and to find the markers of kidney deficiency related to the decrease of ovarian reserve function Can reduce the mechanism to provide research basis.Methods:Experiment 1:To screen out the patients with DOR of kidney deficiency and the infertile women with NOE caused by male factor,and to screen the miRNA of different expression between them by miRNA sequencing technology.Experiment 2:The patients in the treatment group were treated with Erzhi Tiangui granule+IVF,and the patients in the control group were treated with placebo+IVF.Before and after treatment,the changes of TCM syndrome integral,the days of using gonadotropin(GN),the dosage of GN,the estradiol(E2)and progesterone(P)on the day of HCG injection,the number of oocytes obtained,2PN,the number of embryos formed,the number of high-quality embryos and the rate of high-quality embryos were statistically analyzed.The expression of specific miRNA in ovarian granulosa cells was determined by time-lapse fluorescence quantitative analysis(qPCR).Results:Experiment 1:Compared with NOR women,the expression of 45 miRNAs(miR-221-3p,miR-146b-5p,miR-378a-3p,miR-143-5p,miR-222-5p,miR-221-5p)was down regulated(P<0.05),and the expression of 25 miRNAs(miR-6881-3p,miR-4787-3p,miR-4745-5p,miR-6513-3p,miR-3179)was up regulated(P<0.05).The main related pathways are PI3K/Akt and MAPK signal pathways.Experiment 2:1.After treatment,the TCM syndrome score of kidney deficiency syndrome in the treatment group was significantly lower than before(P<0.001).There was no significant difference in TCM syndrome score of kidney deficiency syndrome in the control group before and after treatment(P>0.05).Compared with the control group,before and after treatment,the difference of TCM syndrome score of kidney deficiency syndrome in the treatment group was more significant(P<0.001).2.Compared with the control group,the dosage of GN in the treatment group was significantly reduced,and the level of E2 on HCG day was relatively high,the difference was statistically significant(P<0.05).Compared with the control group,the number of eggs,high-quality embryos and the rate of high-quality embryos in the treatment group increased significantly(P<0.05),but there was no significant difference between the two groups in the number of 2PN and the number of embryos(P>0.05).3.Compared with the control group,the expression of miR-146b-5p,miR-378a-3p,miR-143-5p,miR-222-5p,miR-221-5p and miR-221-3p in the ovarian granulosa cells of the treatment group was up-regulated,and the expression of miR-6881-3p and miR-4787-3p was down regulated,the difference was statistically significant(P<0.05).Conclusion:1.Compared with NOR women,the expression of miR-146b-5p,miR-378a-3p,miR-143-5p,miR-222-5p,miR-221-5p,miR-221-3p,miR-6881-3p and miR-4787-3p in ovarian granulosa cells were significantly different.This difference may be related to the syndrome of kidney deficiency.2.Erzhi Tiangui granule can improve the syndrome of kidney deficiency in DOR patients with kidney deficiency,increase the number of oocytes obtained,the number of high-quality embryos and the rate of high-quality embryos,and affect the expression of 8 specific miRNAs obtained in Experiment 1,which indicates that Erzhi Tiangui granule may improve the syndrome of kidney deficiency by regulating the expression of 8 miRNAs,so as to improve the possibility of pregnancy.3.MicroRNA-146b-5p,miR-378a-3p,miR-143-5p,miR-222-5p,miR-221-5p,miR-221-3p,miR-6881-3p and miR-4787-3p may be markers of kidney deficiency in DOR,providing theoretical basis for further study on the mechanism of kidney deficiency of DOR.