[Effects of Yiqi Bushen Experimental Recipe on mRNA Expressions of foxp3, STAT5, and NF-KB in Decidua Tissue of Natural Abortion Model Mice].

Objective To observe the effects of Yiqi Bushen Experimental Recipe (YBER, a rec- ipe for benefiting qi and Shen supplementing) on mRNA expression of foxp3 in splenic CD4 + CD25 Treg cells and mRNA expressions of foxp3, STAT5, and NF-κB in decidua tissue of natural abortion (NA) model mice. Methods Female CBA/J mice were caged and mated to male DBA/2 mice or male BALB/c mice in 2:1 for NA model. Pregnant CBA/J mice were randomly divided into 5 groups, i.e., the negative control group, the positive control group, high, middle, and low dose YBER groups, 10 in each group. Mice in the NS control group were administered with normal saline by gastrogavage from day 0 to their death, 10 mg/kg, once per day. Mice in the positive control group were administered with Cyclosporine A solution by gastrogavage on the 4th day of pregnancy. YBER (48, 24, 12 g/kg) was respectively administered to mice in the 3 dose YBER groups by gastrogavage from day 0 to their death, once per day. Preg- nant mice were sacrificed at day 9 and 14, and fresh spleens were taken out for extracting Treg cells. Dcidua tissues were collected and stored in -80 °C for frozen. Splenic CD4 + cells CD25 + were isolated and purified by magnetic bead. The purity of CD4 + cells CD25 + was identified by flow cytometry (FCM) before and after magnetic bead. mRNA expressions of foxp3, STAT5A, STAT5B, and NF-KB in decidua tissue were analyzed by RT-PCR. Results The purity of CD4 Treg CD25 could arrived at 88% plus. Its activity could be over 95% after trypan blue test. The average ratio of CD4 CD25+/CD4 was 13. 20% before purified isolation, while it was 91. 43% after purified isolation. Compared with the negative control group, foxp3 mRNA expression level in Treg cells was obviously elevated in the positive control group and the high dose YBER group (P <0. 05). foxp3 mRNA expression level in Treg cells was obviously ele- vated more in the high dose YBER group than in the middle dose YBER group and the low dose YBER group (P <0.05). Compared with the negative control group, mRNA expression levels of foxp3 and STAT5B in decidua tissue increased in the positive control group, high and middle dose YBER groups (P <0. 05). mRNA expression level of STAT5A increased in the positive control group and the high dose YBER group at day 9 and 14; as well as in the middle dose YBER group (P <0. 05, P <0. 01). NF-κB mR- NA expression level in decidua tissue was reduced in the positive control group and 3 dose YBER groups (P<0. 01). Conclusion YBER could up-regulate the expression of foxp3 mRNA in splenic CD4 + CD25 + Treg cells and mRNA expressions of foxp3 and STAT5 in decidua tissues of NA model mice, down-regulate NF-κB mRNA expression in maternal-fetal interface, and promote the maintenance of immune tolerance state.