The Effect Of Erzhi Tiangui Granule On Pregnancy Outcome Of Patients With Repeated Implantation Failure Of Kidney Deficiency Type Based On PD-1 And Th17/Tregs Immune Balance

Objective: To investigate the expression of PD-1,ROR-γt and Foxp-3 in endometrium and the content of s PD-1 and Th17/Treg balance related cytokines in peripheral blood during embryo implantation window in patients of repeated implantation failure(RIF)with kidney deficiency,and preliminarily explore the relationship between the expression of PD-1 and Th17/Tregs and kidney deficiency RIF,in order to provide new ideas for the early diagnosis and treatment of RIF patients.Based on this,to explain the possible mechanism of Erzhi Tiangui Granule in the treatment of RIF with kidney deficiency.Methods: Part I: A total of 20 patients were included in the study,of whom 10 patients with RIF of kidney deficiency type were included in the RIF group and 10 patients with successful pregnancy at the first embryo transfer were included in the control group.Endometrial and peripheral serum were collected at the time of embryo implantation.The expression of PD-1,Foxp-3 and ROR-γt m RNA and protein in endometrium was detected by quantitative real-time polymerase chain reaction(q RT-PCR),Western blot and immunohistochemical techniques,and the contents of s PD-1,TGF-β,IL-17 and IL-10 in peripheral serum were detected by enzyme-linked immunosorbent assay(Elisa).Part II: 66 patients with RIF of kidney deficiency type were randomly divided into TCM group and placebo group,33 cases in each group.Conventional FET was performed after 3 cycles of Erzhi Tiangui Granules,and the placebo group was also treated with conventional FET after 3 cycles of placebo.The changes of kidney deficiency score before and after treatment,biochemical pregnancy rate,implantation rate and clinical pregnancy rate were observed and compared between the two groups.Peripheral serum was collected from patients in both groups on the day of embryo transfer,and their contents of s PD-1,TGF-β,IL-17,and IL-10 were measured and compared by Elisa.Results: Part I: 1.Compared with the control group,the m RNA and protein expressions of PD-1 and Foxp-3 in the endometrium of patients in the RIF group were significantly decreased,while the m RNA and protein expressions of ROR-γt were dramatically increased(P<0.05);2.Compared with the control group,the concentrations of s PD-1 and IL-17 in the peripheral blood serum of patients in the RIF group were significantly increased,but the concentrations of TGF-β and IL-10 were significantly decreased(P<0.05);3.The ratios of TGF-β/IL-17 and IL-10/IL-17 in the peripheral blood serum of patients in the RIF group were significantly lower than those in the control group,and the concentration of s PD-1 was negatively correlated with the ratios of TGF-β/IL-17 and IL-10/IL-17,and the differences were statistically significant(P<0.05).Part II: 1.compared with the placebo group,the scores of patients in the TCM group were significantly lower(P<0.05);2.The embryo implantation rate and clinical pregnancy rate of patients in the TCM group were significantly higher than those in the placebo group,the differences were statistically significant(P < 0.05).But we can not see the significant difference in the biochemical pregnancy rate between the two groups(P> 0.05);3.In the TCM group,the serum concentrations of s PD-1 and IL-17 decreased,the concentrations of TGF-β and IL-10 increased,and the ratios of TGF-β/IL-17 and IL-10/IL-17 were significantly higher than in the placebo group,and the differences were statistically significant(P< 0.05).Conclusions: 1.The immune imbalance at the maternal-fetal interface of RIF patients with kidney deficiency type is abnormal,and the Th17/Treg balance shifts to Th17 that is not conducive to pregnancy,which may be related to the decreased m RNA and protein expression of PD-1 in the endometrium;2.The concentrations of s PD-1 and IL-17 in peripheral serum of RIF patients with kidney deficiency increased,the concentrations of TGF-β and IL-10 decreased during embryo implantation.The concentration of s PD-1 was negatively correlated with the ratios of TGF-β/IL-17 and IL-10/IL-17.This indicate that the peripheral immune environment of RIF patients with kidney deficiency changed,and the increased s PD-1 caused abnormal PD-1/PD-L1 pathway,resulting in the breaking of Th17/Treg balance,which was not conducive to embryo implantation and pregnancy maintenance;3.Erzhi Tiangui Granule can obviously improve the symptoms of kidney deficiency in patients with RIF of kidney deficiency type,and improve their implantation rate and clinical pregnancy rate;4.Erzhi Tiangui Granule,the traditional Chinese medicine for tonifying kidney,may enhance the immune tolerance of patients by reducing the concentration of s PD-1 in peripheral serum and regulating the Th17/Treg immune balance of patients,thereby improving clinical outcomes.