Clinical Study Of Bushen Jianpi Recipe In The Treatment Of Recurrent Spontaneous Abortion And The Mechanism Of Regulating HLA-G-mediated Immunity

Objective To observe the effect of Bushen Jianpi Recipe on the indexes and clinical symptoms of recurrent spontaneous abortion patients with deficiency of spleen and kidney,so as to improve the curative effect of clinical treatment of recurrent spontaneous abortion;To study the effect of serum containing Bushen Jianpi Recipe on trophoblast in vitro,and to explore the mechanism of traditional Chinese medicine regulating HLA-G-mediated maternal and fetal immunity,so as to lay a cellular experimental foundation for the treatment of recurrent spontaneous abortion with traditional Chinese medicine.Methods 1.clinical research:90 cases of early pregnancy patients with recurrent spontaneous abortion were randomly divided into two groups,45 cases in each group.The control group was treated with didroxyprogesterone orally,10mg every time,three times a day,40 days to 12 weeks or more than the previous pregnancy;In the treatment group,on the basis of oral didroxyprogesterone,the traditional Chinese medicine of Bushen Jianpi Recipe was added,One dose per day,two times in the morning and evening,40 days to 12 weeks or more.To observe the success rate of 12 weeks gestation,the improvement of symptoms and signs,the changes of serum indexes before and after treatment(Include the Percentage of E2、P、HCG、sHLA-G、INF-y、IL-2、IL-4、IL-6、IL-10、IL-17A、TNF、Peripheral blood NK cells(CD56+CD16+))and the results of color Doppler ultrasonography.2.experimental study:In this study,HTR-8/svneo trophoblast cell line and NK-92 cell line were selected as cell models in vitro to prepare rat serum containing medicine,and HTR-8/svneo trophoblast and NK-92 cells were cultured,And co culture the two kinds of cells,To observe the changes of HLA-G mRNA,HLA-G protein and the expression of sHLA-G,E2,P,hCG,inf-γ,IL-2,IL-4,IL-17 and IL-10 in the supernatant of trophoblast after cultured HTR-8 cells with different concentrations of serum containing drugs for 48 hours,Changes of invasion and migration of trophoblast in HTR-8 cells cultured with different concentrations of serum for 48 hours,Changes of KIR2DL4 receptor,KIR2DL4 mRNA and KIR2DL4 protein expression in NK cells cultured with different concentrations of serum for 48 hours,Apoptosis of HTR-8+NK-92 cells cultured in different concentrations of serum for 16 hours.Results 1.clinical research:The success rate of the treatment of recurrent spontaneous abortion of spleen kidney deficiency type with Bushen Jianpi Recipe is 76.67%,which can significantly improve the symptoms and signs of spleen kidney deficiency and improve the pregnancy outcome.2.experimental study:The serum containing Bushen Jianpi Recipe can up regulate the expression of HLA-G mRNA,HLA-G protein,E2,P,hCG,IL-4,down regulate the expression of INF-γ,IL-17,promote the migration and invasion of trophoblast;Serum containing Bushen Jianpi Recipe can promote the expression of KIRIDL4 mRNA and protein on the surface of NK cells,and up regulate the expression of KIRIDL4 receptor;Serum containing Bushen Jianpi formula can inhibit the killing effect of NK cells on trophoblast and reduce trophoblast apoptosis.Conclusion 1.The application of Bushen Jianpi Recipe in the treatment of recurrent spontaneous abortion of spleen kidney deficiency type has a higher clinical pregnancy success rate than that of didroxyprogesterone group alone,which can significantly improve the symptoms and signs of patients,with definite curative effect and small side effects,and plays an irreplaceable role in the treatment of recurrent abortion.2.The application of serum containing Bushen Jianpi Recipe in vitro can up regulate the expression of HLA-G in trophoblast,inhibit the killing effect of NK cells,bias the maternal fetal immune response to Th2,promote the invasion and migration of trophoblast,and reduce trophoblast apoptosis.