This study examined the effects of Bangdeyun on the expressions of nuclear factor-kappaB （NF-κB）, interferon-gamma （IFN-y） and interleukin-10 （IL-10） in the endometrium of mice with embryo implantation dysfunction （EID） during the implantation time （namely on pregnancy day 5, 6, 7 and 8） and explored the local immune regulatory effects of Bangdeyun. The gestational mice were randomly divided into normal group, model group and Bangdeyun-treated group. EID models of mice were established by using indomethacin. The endometrial expression of NF-κB was detected by immunohistochemistry and Western blotting. IFN-γ and IL-10 were measured by enzyme-linked immunosorbent assay （ELISA）. The results showed that in the normal group, NF-κB and IFN-γ were weakly expressed and IL-10 was strongly expressed in the endometrium during the whole implantation period. In the model group, the expressions of NF-κB and IFN-T were increased on pregnancy day 5, 6 and 7, and IL-10 expression decreased during the whole implantation time when compared with those in the normal group （P〈0.01 for all）. In the Bangdeyun-treated group, little amount of NF-κB and IFN-γ was expressed and IL-10 expression was strong, much the way they were expressed in the normal group （P〉0.05）. The expressions of NF-κB and IFN-T were much lower in the Bangdeyun-treated group than those in the model group on pregnancy day 5, 6 and 7 （P〈0.01 for all）, while the expression of IL-10 was much higher than in the model group during the whole implantation time （P〈0.01）. It was suggested Bangderun may favor a shift from Thl- to Th2-type immune response, therefore inhibiting the maternal immune rejection, inducing the immune tolerance and improving the fetal implantation.
Summary： The study examined the effect of DS147, the bioactive component of the traditional herbal recipe Bangdeyun, on pregnancy in mice with embryo implantation dysfunction induced by controlled ovarian stimulation （COS）, and the underlying mechanisms. Female mice were superovulated by intrap eritoneal injection of 7.5 IU of pregnant mare serum gonadotropin （PMSG） followed by an additional injection of 7.5 IU hCG 48 h later to establish embryo implantation dysfunction （EID） model. Pregnant mice were randomly divided into normal control group, COS group and DS147-treated groups. The pregnancy rate and the average implantation site were obtained on pregnancy day 8 （PD8）. The side effect of 200 mg/kg of DS147 on naturally pregnant mice was also observed. Further, the uterine and ovarian tissue samples were collected on PD5 for measuring their weights, observing the development of the endometrium and ovary, and detecting the endometrial expression ofMMP-2, TIMP-2, CD34 and angiogenin （ANG）. The female mice treated with DS147 at doses of 100 to 800 mg/kg showed a higher pregnancy rate than those in COS group, and the highest pregnancy rate of 83.3% occurred in the 200 mg/kg DS147-treated group. Moreover, no obvious side effect was found in mice treated with 200 mg/kg DS147 on PD8 and PD16. The ovarian and uterine weights, and the expression levels of MMP-2, ANG and CD34 were significantly increased in DS147-treated groups when compared with COS group. The TIMP-2 expression level was much lower in DS147-treated mice than in COS mice and the ratio of MMP-2/TIMP-2 was much higher in DS147-treated group than in COS group, and even higher than normal control group. In all, these findings suggest that DS147 may improve pregnancy in mice with COS-induced EID by promoting matrix degradation and angiogenesis, and improving the development of corpus luteum and endometrial decidualization around the implantation window.
