| ObjectiveExplore transforming growth factor-?1(transforming growth factor-?1,TGF-?1),CD4 + CD25 + regulatory T cells(Treg),forkhead or winged helix transcription factor 3(forkhead or winged helix transcription,FOXP3)Cellular immune regulation mechanism;observe the changes of peripheral blood TGF-?1,Treg and FOXP3 expression levels before and after treatment in patients with kidney deficiency,liver stagnation and blood stasis by multiple interventions of traditional Chinese medicine,and compare the differences in expression levels before and after treatment,and Analyze the correlation between the changes of TGF-?1,Treg,FOXP3 expression levels and clinical efficacy,explore the multi-path intervention of Chinese medicine to regulate the immune tolerance mechanism of patients with IVF-ET long protocol,and provide theoretical support for Chinese medicine clinical pregnancy.Methods1.According to the inclusion criteria and exclusion criteria,20 patients with kidney deficiency and liver-qi depression combining blood stasis syndrome planned to undergo IVF-ET long protocol after the failure of the long protocol of Huaxi Second Hospital of Sichuan University from September 2016 to February 2018 were divided into treatments Group and natural waiting group,each group have 10 cases.The treatment group was treated with TCM orally,acupressure,and Chinese medicine retention enema for 3 months in the Department of TCM and Gynecology of the Affiliated Hospital of Chengdu University of Traditional Chinese Medicine.The natural waiting group included in the same period naturally waited for 3 months after the failure of the long protocol.Patients in the treatment group received 5ml of fasting venous blood before intervention and 3 months after intervention,that is,6-7 days after LH peak [d(LH + 6)],P?5ng / ml,and Gn RH-a;In the natural waiting group,5 ml of fasting venous blood was drawn at the same time point after IVF-ET failure and natural waiting for 3 months.2.ELISA method was used to detect the expression of TGF-?1 and FOXP3 protein in peripheral blood;real-time fluorescent PCR was used to detect the expression of TGF-?1 m RNA and FOXP3 m RNA in peripheral blood;Treg cell expression was analyzed by flow cytometry,and it was detected on Excel software After finishing the data entry and sorting,the data entered was statistically compared and analyzed using SPSS 25.0 statistical software.Using blank control and self-pre and post-control methods,observe the natural conception,pregnancy outcome,and TCM syndrome scores of patients in the two groups before IVF-ET,peripheral blood TGF-?1,FOXP3,and Treg cells before and after the patient and between the two groups The changes of expression levels were analyzed,and the correlation between the changes of their expression levels was analyzed to explore the mechanism of TCM intervention through the method of tonifying the kidney,replenishing the liver and promoting blood circulation to regulate the immune tolerance and assist pregnancy in IVF-ET patients.Results1.Natural pregnancy rate: 10 cases of IVF-ET failure in each of the two groups who plan to undergo IVF-ET long-term regimen again,3 cases of natural pregnancy during the intervention period in the treatment group,and 0 cases of natural pregnancy before the IVF-ET treatment in the natural waiting group.The natural pregnancy rate in the treatment group was higher than that in the natural waiting group(P <0.05).2.Repeated IVF-ET pregnancy outcome: 7 patients in the treatment group underwent IVF-ET long course treatment,of which 3 cases were successfully implanted with IVF-ET;naturally waiting group 10 cases underwent IVF-ET long course treatment,2 cases with IVF-ET implantation was successful,including a biochemical pregnancy.The clinical pregnancy rate was higher in the treatment group than in the natural waiting group(P <0.05).3.TCM Syndrome Points and Efficacy: The TCM Syndrome Points of kidney deficiency and liver-qi depression combining blood stasis syndrome of Patients in the Treatment Group After Multi-channel Intervention Decreased Compared with Before the Intervention(P <0.05);Patients in the Natural Waiting Group did not receive TCM intervention before and after,There was no significant difference in the scores of syndromes of kidney deficiency and liver-qi depression combining blood stasis syndrome(P>0.05).After three months of intervention /waiting between the two groups,compared with the two groups,the total effective rate of TCM syndromes in the treatment group was significantly higher than that in the natural waiting group(P <0.05).4.TGF-?1: The expression levels of TGF-?1 protein and TGF-?1 m RNA in the treatment group were significantly higher than those before the intervention(P <0.05);the natural waiting group did not receive the TGF-?1 protein,TGF-?1m RNA expression levels were not significantly different(P>0.05);after two months of intervention / waiting in the two groups,compared between the two groups,the TGF-?1m RNA expression level in the treatment group was higher than that in the natural waiting group after intervention(P<0.05)).5.FOXP3: Compared with the multi-channel intervention of Chinese medicine in the treatment group,the expression levels of FOXP3 protein and FOXP3 m RNA were significantly higher after intervention than before intervention(P <0.05).There was no significant difference in the expression levels of FOXP3 protein and FOXP3 m RNA before and after TCM intervention in the natural waiting group(P>0.05).After three months of intervention / waiting between the two groups,the FOXP3 m RNA expression level in the treatment group was higher than that in the natural waiting group(P<0.05),and there was no significant difference in FOXP3 protein expression(P>0.05).6.Treg: The Treg expression level in the treatment group after three-month intervention of TCM was significantly higher than that before the intervention(P <0.05);there was no significant difference in Treg cell expression levels before and after waiting in the natural waiting group(P> 0.05);After three months of intervention / waiting in the group,the Treg expression in the treatment group was higher than that in the natural waiting group after intervention(P <0.05).7.There was a positive correlation between TGF-?1 and Treg expression levels(r = 0.649,P = 0.042),changes in TCM syndrome scores and FOXP3 values(r =-0.363,P = 0.021),Treg values(r =-0.334,P = 0.035)were negatively correlated.8.The expressions of TGF-?1,FOXP3,and Treg in peripheral blood of 3 patients with natural pregnancy were increased after multi-channel intervention of Chinese medicine;the expression levels of TGF-?1,FOXP3 and Treg increased in 5 cases before and after intervention/waiting-The proportion of patients with successful ET implantation was 80.0%,80.0%,and 100%,respectively,and 87.5%,87.5%,and 100% of the 8 pregnant patients.The change trend of TGF-?1 and Treg and the pregnancy outcome were statistically significant(P <0.05).ConclusionMulti-channel intervention of traditional Chinese medicine in patients with kidney deficiency and liver-qi depression combining blood stasis syndrome IVF-ET long plan can improve patients' pregnancy outcomes and clinical symptoms related to kidney deficiency and liver-qi depression combining blood stasis syndrome;preliminary conclusions have been drawn that the mechanism may be based on the method of tonifying the kidney,stagnation and blood circulation TCM multi-path treatment can up-regulate the expression level of TGF-?1 in peripheral blood of IVF-ET long protocol patients,induce the expression of "nuclear transcription factor,an important regulatory gene" FOXP3 in Treg cells,and promote the differentiation of primary CD4+T cells into Treg The cells can expand the expression of local Treg cells at the maternal-fetal interface,mediate immune tolerance,exert immunosuppressive effects,protect semiallografts from maternal immune attack and implant smoothly,Thereby helping the bed. |
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