Based On SDF-1??E₂ And CXCR-4 Expression,The Effect Of Bushenantai Granule On The Homing Mechanism Of EPCs In Kidne Y-deficiency Abortion Model Was Studied

1 Objective On the basis of previous studies,this clinical study intends to study the clinical efficacy of Bushenantai granule and its effect on maternal-fetal interfacial angiogenesis by taking the contents of E₂ and SDF-1? in peripheral blood as the entry point,so as to provide theoretical support for the further study of the role of tonifying kidney and strengthening spleen traditional Chinese medicine in the homing mechanism of endothelial progenitor cells in mice with kidney deficiency abortion.2 Methods From January 2019 to December 2019,30 patients with kidney deficiency abortion treated by Bushenantai granule were selected as the study group according to the standard among the gynecological outpatient patients of the first affiliated hospital of anhui university of traditional Chinese medicine,and the contents of SDF-1? and E₂ were detected in peripheral blood at the time of treatment and 2 weeks after treatment.During the same period,30 patients in the normal first trimester were selected as the blank group,and the SDF-1? and E₂ levels were detected in the peripheral blood at the time of treatment and 2 weeks later.The results were statistically analyzed.3 Results3.1 Comparision of Characteristics between two groups There was no significant difference in age,gestational age and number of miscarriage between the two groups?P>0.01?.3.2 Clinical efficacy of study group The study group treated 12 cases,10 cases were effective,and 8 cases were ineffective.The effective rate is 73.3%.3.3 The two groups of detection indicators were compared3.3.1 Comparison of SDF-l? in peripheral blood?1?At the first diagnosis: the content of SDF-1? in peripheral blood of the blank group was higher than that of the study group,and the difference was statistically significant?P<0.01?;?2?After 2 weeks: the content of SDF-1?in the peripheral blood of the blank group increased significantly compared with that of the first diagnosis after 2 weeks?P <0.01?;the content of SDF-1?in the peripheral blood of the study group increased significantly after the drug administration compared with that before the drug administration,and the difference was statistically significant?P<0.01?;there was no difference in SDF-1?levels in peripheral blood of the study group and the blank group after 2 weeks?P>0.01?.3.3.2 Comparison of E₂ in peripheral blood?1?At the first diagnosis:the content of E₂ in the blank group was higher than that in the study group,and the difference was statistically significant?P<0.01?.?2?After 2 weeks:the content of E₂ in peripheral blood of the blank group increased significantly compared with that of the first diagnosis,and the difference was statistically significant?P < 0.01?;the content of E₂ in peripheral blood of the study group increased significantly after the drug administration compared with that before the drug administration,and the difference was statistically significant?P< 0.01?;the level of E₂ in peripheral blood of the study group was lower than that of the blank group2 weeks later,and the difference was statistically significant?P<0.01?.3.3.3 Correlation between E₂ and SDF-l? in peripheral blood There was a positive correlation between E₂ and sdf-l in peripheral blood of blank group and study group?P<0.01?.4 Conclusion4.1 The content of SDF-l? and E₂ in patients with spontaneous abortion was lower than that in patients with normal pregnancy,suggesting that the content of SDF-l? and E₂ was correlated with spontaneous abortion.4.2 The contents of SDF-l? and E₂ in peripheral blood were positively correlated and might increase with the growth of gestational weeks.4.3 Bushenantai granule may improve the clinical symptoms of patients with spontaneous abortion by upregulation of SDF-l?.1 Objective On the basis of clinical research,this part of the experiment intends to study the effects of Bushenantai granule on the mobilization and homing mechanism of EPCs at the maternal and foetus interface of aborted mice based on the embryo loss rate,number of EPCs,SDF-1? and CXCR-4 content.To study the effect of Bushenantai granule on the mobilization and homing of EPCs at the maternal fetal interface of aborted mice and its related mechanism,and further reveal the significance of kidney-tonifying and spleen-tonifying traditional Chinese medicine on the vascular recast at the maternal fetal interface of aborted mice,so as to provide theoretical support for the intervention of traditional Chinese medicine in natural abortion.2 Methods2.1 Establishment of model2.1.1 Bone marrow transplantation model?1?Modeling process: six GFP transgenic mice with stable systemic expression of green fluorescent protein were selected as donor mice to prepare bone marrow mononuclear cells.A total of 100 female ICR mice were used as the recipient mice.After the whole body X-ray irradiation,the recipient mice were injected with GFP-labeled bone marrow mononuclear cells from the tail vein to establish the bone marrow transplantation model.?2?Modeling test: after 4 weeks of bone marrow reconstruction,peripheral blood was collected to detect the proportion of GFP-positive cells,and the mice with successful BMT model were selected for further modeling.2.1.2 Establishment of abortion model of kidney deficiency Hydroxycarbamide?450mg/kg.d?was administered for modeling from day 1 to day 9 of gestation.Mifepristone?4.0mg/kg.d?was administered in the morning of the 10 th day of gestation to replicate the kidney failure abortion model.2.2 Experimental grouping and administration40 GFP-labeled BMT mice with kidney failure were randomly divided into 4 groups with 10 mice in each group.In addition,10 GFP-labeled BMT mice were selected as the blank group.In this study,the clinical application dose of Bushenantai granule was converted into the dose of mice according to the calculation method of body surface area and the clinical application dose of Bushenantai granule.10 GFP-labeled BMT mice were given normal saline by gavage as a blank group.GFP-labeled BMT kidney deficiency abortion mice in 10,normal saline gavage,as the model group.10GFP-labeled BMT mice with kidney deficiency abortion were treated with Bushenantai granule?3.0g/kg.d?by gavage as a low-dose group.GFP-labeled BMT mice with kidney deficiency abortion were administered by gavage with Bushenantai granule?6.0g/kg.d?as the medium dose group.GFP-labeled BMT mice with kidney deficiency abortion were administered by gavage with Bushenantai granule?12.0g/kg.d?as a high-dose group of traditional Chinese medicine.2.3 Sampling and index detection2.3.1 Bone marrow Flow cytometry was used to detect the number of endothelial progenitor cells.2.3.2 Peripheral blood?1?Flow cytometry was used to detect the number of EPCs cells in peripheral blood of mice.?2?ELISA was used to detect SDF-1? in peripheral blood.2.3.3 Decidual tissues?1?Flow cytometry was used to detect the number of EPCs cells in mouse decidual tissues.?2?The expression level of CXCR-4 was detected by ELISA.3 Results3.1 Detection of bone marrow chimerism rate of GFP Six mice died after bone marrow transplantation,and 50 mice were successfully modeled.The modeling success rate of bone marrow transplantation model was 53.2%.3.2 Abortion rate and embryo loss rate of pregnant mice in each group Abortion rate: blank group = high dose group < medium dose group < low dose group < model group;The embryo loss rate of the blank group was lower than that of the model group and the low-dose group,and the difference was statistically significant?P < 0.01?.The embryo loss rate in the model group was higher than that in the high-dose and medium-dose groups,and the difference was statistically significant?P<0.01?.The embryo loss rate in the model group was higher than that in the low-dose group,and the difference was statistically significant?P < 0.05?.Compared with the high-dose group,the low-dose group had a higher embryo loss rate,and the difference was statistically significant?P<0.05?.Compared with the low-dose group,the embryo loss rate in the medium-dose group was lower,and the difference was statistically significant?P<0.05?.3.3 The number of bone marrow endothelial cells in each group Compared with the blank group,the number of bone marrow EPCs was less in the model group and the high-dose,medium-dose and low-dose groups,and the difference was statistically significant?P<0.01?.Compared with the model group,the number of bone marrow EPCs in the high-dose group was more,and the difference was statistically significant?P<0.01?.Compared with the high-dose group,the number of bone marrow EPCs in the low-dose and medium-dose groups was less,and the difference was statistically significant?P<0.01?.There was no significant difference in the number of bone marrow EPCs between the low-dose group and the medium-dose group?P> 0.01?.3.4 Peripheral blood endothelial cells of pregnant mice in each group Compared with the blank group,the number of EPCs in peripheral blood in the model group,the medium dose group and the low dose group was less,with statistically significant difference?P<0.01?,while the number of EPCs in the high dose group was less,with statistically significant difference?P<0.05?.Compared with the model group,the number of EPCs in peripheral blood of the high-dose,medium-dose and low-dose groups was large,and the difference was statistically significant?P<0.01?.There was no statistical difference in the number of EPCs in peripheral blood between the high,medium and low groups?P>0.01?.3.5 Comparison of SDF-1?in peripheral blood of pregnant mice in each group The content of SDF-1?in the peripheral blood of the blank group was higher than that of the model group and the low-dose group,and the difference was statistically significant?P < 0.01?.The difference was statistically significant?P < 0.05?,and there was no significant difference?P > 0.01?.The content of SDF-1? in peripheral blood of the model group was lower than that of the high-dose,medium-dose and low-dose groups,and the difference was statistically significant?P<0.01?.The content of SDF-1?in the peripheral blood of the low-dose group was lower than that of the high-dose group,and the difference was statistically significant?P<0.05?.There was no significant difference in the content of SDF-1?in peripheral blood in the medium dose group compared with the high dose group?P>0.01?.3.6 Number of endothelial cells in decidual tissues of pregnant mice in each group Compared with the blank group,the number of EPCs was lower in the model group and the high-dose,medium-dose and low-dose groups,and the difference was statistically significant?P < 0.01?.Compared with the model group,the number of EPCs in the high-dose and medium-dose groups was high,and the difference was statistically significant?P<0.01?;the number of EPCs in the low-dose group was high,and the difference was statistically significant?P<0.05?.Compared with the high-dose group,the number of EPCs in the low-dose group was low,and the difference was statistically significant?P<0.01?.The number of EPCs in the medium-dose group was low,and the difference was statistically significant?P <0.05?.Compared with the low-dose group,the number of EPCs in the medium-dose group was higher,and the difference was statistically significant?P<0.05?.3.7 Expression of CXCR-4 in decidual tissues of pregnant mice in each group Compared with the blank group,the CXCR-4 content in the decidual tissues of the model group was less than that of the high,medium and low dose groups,and the difference was statistically significant?P<0.01?.Compared with the model group,the contents of CXCR-4 in the decidual tissues of the high-dose and medium-dose groups were higher,and the difference was statistically significant?P<0.01?.The contents of CXCR-4 in the decidual tissues of the low-dose groups were higher,and the difference was statistically significant?P<0.05?.There was no significant difference in CXCR-4content in decidual tissues between groups with high,medium and low dose of traditional Chinese medicine?P>0.01?.4 Conclusion4.1 Bushenantai granule has an effect on the abortion rate of mice and can reduce the embryo loss rate of mice.The effect may be related to the concentration.4.2 Compared with aborted mice,normal pregnant mice had higher SDF-1 content in peripheral blood,higher CXCR-4 content in decidual tissues,and more EPCs in bone marrow,peripheral blood and decidual tissues.4.3 The amount of EPCs in bone marrow,peripheral blood and decidual tissues could be increased by Bushenantai granule.4.4 Bushenantai granule may promote the mobilization of EPCs by upregulation of SDF-1? in peripheral blood,and the expression of CXCR-4 in mouse decidual tissues can promote the homing of EPCs.