| Background and ObjectiveNowadays, with the increasing number of infertility patients year by year, assisted reproductive technology(ART) emerged as the times require, which opened up a new field for the clinical treatment of infertility and brought a new hope to infertility patients. Although ART has been progressed for more than three decades, there is still a phenomenon which is high fertilization rate and low pregnancy rate. Study has been found that two-thirds patients who were failed with in vitro fertilization-embryo transfer(IVF-ET) treatment are due to the decline in endometrial receptivity. Therefore, it is crucial to improve endometrial receptivity so as to enhance implantation rate and clinical pregnancy rate. Endometrial receptivity is the ability of endometrium to accept embryo. Normally, the endometrium allow the embryo implant in the endometrium for a limited time when is called the implantation window or nidation window, that is from the twentyth to twenty-fourth day during the normal menstrual cycle, or the fourth day after pregnancy of mice.Recently, many molecular factors have been found to participate in embryo implantation process. Leukaemia inhibitory factor(LIF), integrinβ3and homeobox A10(HOXA10) are the most typical ones, whose expression peak period are consistent with endometrial implantation window. Furthermore, decline, deletion or mutation of them may result in infertility. So LIF, integrinβ3and HOXA10are considerd as markers of uterus receptivity.Superovulation and controlled ovarian hyperstimulation(COH) are the important step of ART treatment, but many studies have shown that they can reduce endometrial receptivity and prohibit embryo from implanting. In the field of reproductive medicine, how to improve endometrial receptivity has been urgent issue to tackle. Many domestic and international scholars have carried on pharmaceutical researches to improve endometrial receptivity, which focus on estrogens, low-dose aspirin, growth hormone and traditional Chinese prescriptions. However, these duges could not establish perfect endometrial implantation microenvironment and have some drawbacks. For instances, the security of estrogens and aspirin has been questioned. The price of growth hormone is too high. Besides, it is inconvenient to take Chinese medicinal herb and the infertility patients have poor medication compliance. For these reasons, it is imperative to search for a pharmaceutical which is safe, effective, convenient and cheap. Under the background, it is by the immunohistochemical streptavidin-peroxidase(SP) technique that we observe the effect of Kuntai capsules on expressions of LIF, integrinβ3and HOXA10in endometrium of natural cycle, superovulation cycle and COH cycle during the implantation window of mice. The purpose is to provide a new medicine selection and theoretic basis for improving the endometrial receptivity by investigating effects of Kuntai capsules on endometrial receptivity and its mechanism.Materials and MethodsNinety mice were randomly separated into six groups:normal saline(NS) group(A group), NS plus Kuntai capsules group(B group), superovulation protocol group (C group), superovulation protocol plus Kuntai capsules group (D group), COH protocol group (E group), COH protocol plus Kuntai capsules group (F group). Animal models were established, which were referring to the relevant literatures. All the mice were put to death on the pregnant fourth day and the uterus samples were collected. The expressions of LIF, integrin(33and HOXA10in endometrium were detected and analysed semi-quantitatively by immunohistochemical SP method. The data were statistically analysed by SPSS17.0software. Comparison of the mean was performed by student t-test in two groups and by one-way analysis of variance among multigroups. Multiple comparison was accomplished by least significant difference(LSD)t-test.Results1. Immunostaining of LIF and integrinβ3protein are primarily located in cytoplasm of uterus luminal epithelial cells and uterus glandular epithelial cells during nidation window. There is also a little expression could be seen in stromal cells. While HOXA10protein is expressed in uterus luminal epithelial cells, uterus glandular epithelial cells and stromal cells.2. Compared with normal saline group, the standard of LIF, integrin(33and HOXA10protein are significantly rised in NS plus Kuntai capsules group (P<0.05). Compared with superovulation protocol group, the level of the three cytokines are significantly increased in superovulation protocol plus Kuntai capsules group (all P<0.001). The expressions of three proteins in COH protocol plus Kuntai capsules group are statistically higher than that in COH protocol group (all P<0.001).3. The level of LIF, integrinβ3and HOXA10protein in control group are statistically higher than that in superovulation protocol group (all P<0.001) and COH protocol group (all P<0.001). In comparison with superovulation protocol group, the level of the three proteins are significantly increased in COH protocol group (all P<0.05).4. The level of LIF, integrinβ3and HOXA10protein in control group are statistically higher than that in superovulation protocol plus Kuntai capsules group (all P<0.001) and COH protocol plus Kuntai capsules group (P<0.01). Compared with superovulation protocol plus Kuntai capsules group, the level of LIF and integrinβ3protein are statistically increased in COH protocol plus Kuntai capsules group (P<0.01).Conclusions 1.Immunostaining of LIF, integrinP3and HOXA10protein are positive in the endometrium of mice during the implantation window. So LIF, integrinβ3and HOXA10are seen as markers of endometrial receptivity.2. Superovulation protocol and COH protocol can both impair endometrial receptivity, but superovulation protocol has more negative effect on endometrial receptivity than COH protocol.3. Kuntai capsules can partly improve endometrial receptivity of mice in natural cycle, superovulation cycle and COH cycle. |
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