| Baicalin, a flavonoid ingredient extracted from Scutellaria, has been widely applied for promoting blastocyst implantation and keeping pregnancy. Its tocolysis are mainly based on the follows:firstly, Baicalin can regulate the Th1/Th2cytokines balance either directly or indirectly, targeting to establish an environment for blastocyst adhesion. Secondly, Baicalin has effects on the amount of pregnance-related steroid hormones and their receptors, through which to improve conditions of maternal-fetal interface for implantation. And thirdly, Baicalin suppresses the contraction of uterine smooth muscle that caused by various factors in order to protect the early embryo implantation. But the relationship between the anti-abortion effects of Baicalin and the Wnt pathways has not been explored yet. And gravidity is a complex physical process which involves multiple systems and pathways. Therefore, we conduct this experiment to probe into the molecular mechanism of anti-abortion of Baicalin.Virgin Kunming mice were used in this experiment. The males mated the females at the ratio of1:2. If the pessary was found the next morning, the females were determined to begin pregnancy, and recorded as the first day of pregnancy (d1). The gravidity mice were randomly divided into four groups according to giving different drugs:control group, Baicalin group, RU486group, and Baicalin+RU486group. Baicalin were administered to the Baicalin group mice, RU486to the RU486group mice, Baicalin and RU486to the Baicalin+RU486group mice, and solvent to the control mice at d2-8, and euthanized for calculating pregnancy rate and implantation blastocyst number at d8.At d3to8of gestation, blood and uterine tissue samples were taken. Concentrations of progesterone and estradiol were measured with radioimmunoassay, the mRNA and protein levels of beta-catenin, LRP6, Dkkl and Wnt4were detected with qRT-PCR and Western-blotting, respectively.The results showed that Baicalin cause serum estradiol concentration to decline (P<0.05), and suppressed the elevated estradiol levels caused by Mifepristone (P<0.01). Baicalin led to the increase in serum progesterone level (P<0.05), suppressed the decrease in progesterone level caused by Mifepristone (P<0.05) during peri-implantation. Compared with the control, Dkkl mRNA level in uterine tissue decreased at d4-8(P<0.05) while Wnt4, p-catenin and LRP6mRNA increased at d4-8(P<0.05), d3-8(P<0.01) and d3-7(P<0.01), respectively. Western-blotting experiments showed that Baicalin caused beta-catenin protein to increase at d4-8(P<0.05), LRP6to increase at d3-7(P<0.05), and Dkkl to decrease significantly in d6-8(P<0.05) in mouse uterine tissues.In this study, the model of mouse blastocyst implantation dysfunction was established with RU486. The effects of Baicalin on the estradiol and progesterone in mouse serum, and on the mRNA and protein levels of canonical Wnt signaling pathway related substances (β-catenin, LRP6, Dkk1and Wnt4) in the uterine tissue were studied. It was found that Baicalin had some effects on expressing levels of β-catenin, LRP6, Dkkl and Wnt4, suggesting that Baicalin may affect pregnancy in mice through the canonical Wnt signaling pathway. |
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