TJCYR ameliorates embryo implantation dysfunction through PI3K/Akt/eNOS signalling to improve endometrial receptivity

Abstract Backgroud: In our clinical practice, we found that Tiao Jing Cu Yun Recipe (TJCYR), which was composed of Dangshen, Danshen, Danggui, Huangqi, Shudihuang, Bajitian and Yinyanghuo, had obviously enhanced the rate of pregnancy in the women with infertility. Although the application effects are desirable and satisfactory, the therapeutic mechanism of TJCYR remains poorly understood. In this study, we evaluated the effect of TJCYR on embryo implantation dysfunction (EID)-induced damage of endometrial receptivity in mice and investigated the mechanisms underlying the effect. Methods: Pregnant mice were randomly divided into six groups: Control, EID only, Progesterone (Prog)+EID, TJCYR-low-dose+EID , TJCYR-medium-dose+EID, TJCYR-high-dose+EID. Mifepristone was injected to make EID model. On the eighth day of pregnancy, the mice were sacrificed and the number of uterus-implanted blastocysts was counted. On the fourth day of pregnancy, the serum was to analyze the level of hormone by radioimmunoassay, the uterus was to analyze morphology by H&E and SEM, the combination of immunofluorescence and western blot were to identify the related proteins. Results: Compared with the EID group, the mice treated with high-dose TJCYR had a greater number of implanted sites, so we choose that dose of TJCYR as treatment in the following study. The mice treatment with TJCYR could significantly enhance the level of P 4 , and inhibite the decrease in the expression of PR that induced by EID. Compared with the EID only, the SEM showed that a marked increase in the number of well-developed pinopodes in the TJCYR treatment group. Except morphological marker, several molecules in relation to pinopodes that could be used as biomarkers. TJCYR abrogated the EID-induced weakened in those biomarkers. Additionally, the vascular density and VEGF were decreased in the EID group, it appeared severe tissue hypoxia, while TJCYR reversed that change. Compared with the control, the p-Akt and p-eNOS were decreased in EID group, accompanied with decline of NO. While TJCYR promoted the activation of Akt and eNOS, to improve the poor microvascular environment of endometrium. Conclusion: TJCYR has therapeutic potential against poor endometrial receptivity via activation of the PI3K/AKT/eNOS signaling pathway.