| Objective:Embryo implantation is a process in which the endometrium in the receiving state interacted with the embryo in the activated state, and then the trophoblast established close links with the endometrium. The sensitivity of uterus to embryo can be classified into three periods, prereceptive, receptive and refractory. When the endometrium is in the receiving state, that is implantation window, and it allows the embryo to implant in a short time. The process of embryo implantation includes the apposition, attachment, and invasion. However, the endometrial receptivity and embryo activation are two independent events. Only when they developed simultaneously, can the embryo implant. There are many anti-adhesion molecules and adhesion molecules playing a role in the implantation process. Mucin 1 (Muc1) is one of the anti-adhesion molecules. Mucl protein disappeared in the receiving state to allow the embryo implant.Arnebia is a kind of common Chinese herbal medicine, which is commonly used in antifertility and has a long history. It has been demonstrated that arnebia has an anti-fertility effect on a variety of mammals, but the mechanism of function remain unclear. Therefore, in order to elucidate the anti-implantation mechanism of arnebia, we study the affects of arnebia on the implantation process and the Mucl protein' expression by using hematoxylin-eosin (HE) staining, immunohistochemical staining, Western blot in mice.Methods:Two adult females were caged with a male mice or a vasectomized male mice overnight and checked for vaginal plug on the following morning. Presence of vaginal plug was considered as evidence of mating, and the day was designated as the Day 1 postcoitum or the Day 1 pseudopregnancy. (1)The mated female mice were randomly divided into three experimental groups and control group, each group containing 42 mice. The experimental group was fed with arnebia root solution and the control group was fed with saline on day 1, 2,3,4 of pregnancy. The uteri of the mice were removed at 22:00 of on day 4 of pregnancy, one side of the uteri were stained with HE and immunohistochemistry to observe the morphological changes and the expression of mucl protein and the other side of the uteri were measured with western blot to observe the expression of Mucl protein; In the morning of on day 8 of pregnancy, fifteen female mated mice of each group were killed by means of cervical dislocation to observe the rate of embryo implantation and the affects of arnebia on embryo, the number of cubs given birth to by the remaining female mice in each group were observed; 30 days after parturition, the female mice were mated once again and the number of cubs was also taken down. (2) The mated female mice were randomly divided into three experimental groups and control group, each group containing 15 mice. The experimental group was fed with arnebia root solution and the control group was fed with saline on day 11,12,13,14 of pregnancy. In the morning of on day 18 of pregnancy,the mice were killed by means of cervical dislocation to observe the rate of abortion. (3) The pseudopregnancy mice were randomly divided into three experimental groups and control group, each group containing 12 mice. The experimental group was fed with arnebia root solution and the control group was fed with saline on day 1,2,3,4 of pregnancy. The uteri of the mice were removed at 22:00 of on day 4 of pregnancy. One side of the uteri were stained with HE and immunohistochemistry to observe the morphological changes and the expression of mucl protein and the other side of the uteri were measured with western blot to observe the expression of Mucl protein.Results:1. In the experimentⅠ,Ⅱ,Ⅲand the control group, the embryo implantation was successfully conducted in 6,4,1,15 mice respectively; the average number of implanted sites were 6.17±0.79,5.25±0.95,4,10.40±0.52; the anti-implantation rates was 40%,73.3%,93.4%,0 2. In the experimentⅠ,Ⅱ,Ⅲand the control group, the number of pregnant female in the first time was 7,4,1,15; the average number of cubs were 5.28±0.64,3.75±0.48,3,10.54±0.62; the number of pregnant female in the second time was 15,15,15,15; the average number of cubs were 11.11±0.52,9.47±0.57,10.33±0.59,10.54±0.563. In the experimentⅠ,Ⅱ,Ⅲand the control group, the number of pregnant mice was 8,7,4,15; the average implantation sites were 8.38±0.67,7.14±0.51,5.75±0.48,10,33±0.57.4. The endometrium in the experimental group exhibited morphological changes compared with that in control group.5. The experimental results in true pregnant mice showed that the expression of mucl protein was increased with increasing doses of arnebia root, while that in control group showed little increase (P< 0.05). The experimental results in pseudopregnancy mice was seems as in the true pregnancy.Conclusion:1. Arnebia has obvious anti-implantation effect. The number of mice with successful implantation and the average implantation sites were significantly reduced.2. The anti-fertility affect of arnebia was reversible and the fertility of the treated mice can be recovered after using arnebia for a period of time.3. Arnebia has abortion effect, but obviously less than the anti-implantation effect.4. Arnebia can affect the endometrial morphology in some extent5. The arnebia root can prevent the embryo implantation by inhibiting the mucl protein disappearance. |
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