Expression Rule Of MTOR Gene In Endometrium Of Early Pregnant Mouse

Objective:To investigate the molecular mechanism of mTOR during the implantation process, we analyzed the diversity expression of mTOR in the different stages of early pregnant mice endometrium by real-time fluorescence quantitative PCR (FQ-PCR), in-situ hybridization, western blotting, and immunohistochemistry (IHC). This work will be the base to understand further the mechanism of normal reproduction process in human and provide the evidence for diagnosis and treatment of the reproduction aspect disease in human.Methods:1. Found the animal model of NIH pregnant mice. Pregnant mice were randomly divided into 5 experiment groups (pregnant d3, d4, d5, d6 and d7), and the non-pregnant mice were control group, and another was pseudopregnant group. Mice endometria were separated from non-pregnant, pseudopregnant and pregnant mice, and then immediately stored in -80℃for further test.2. The expression of mTOR mRNA was detected by FQ-PCR and in-situ hybridization in the different stages of early pregnant mice endometrium.3. The expression of mTOR was detected by western blotting and immunohistochemistry test in the different stages of early pregnant mice endometrium.Result:1. The result of FQ-PCR indicated that the mTOR mRNA expression in endometrium of pregnant groups was higher significantly than that of non-pregnant group (P<0.05), and the expression of mTOR mRNA appeared that there was a gradual increase from d3 to d5, and reached the maximum level on d4 and d5, and then decreased dramatically on d6 and d7.The mTOR expression increased significantly in the implantation window (P<0.05). Furthermore, the mTOR expression of pseudopregnant group was higher slightly than that of non-pregnant group.2. The result of western blotting showed that the expression pattern of mTOR protein was the same as the result of FQ-PCR.3. The results of in-situ hybridization and immunohistochemistry showed that mTOR expression in endometrial stroma cells was evidently higher than that in endometrial epithelium cells (P<0.05), and the expression pattern of mTOR was the same as the results of FQ-PCR and western blotting. Conclusion:1. The result showed that the expression of mTOR mRNA in pregnant groups was higher significantly than that in non-pregnant group, which indicated that mTOR played a significant role for endometrial cell growth, proliferation, differentiation.2. mTOR expression amount was kept on increasing from the preimplantation period to the implantation window, and reached the extreme on the implantation window d4 and d5. The results might indicate that mTOR, as a central controller, might amplify the activity of signaling pathway for promoting protein synthesis and energy release and further propose a role in providing energy and substance in order to promoting endometrial cells growth, proliferation and differentiation.3. After implantation, the mTOR expression on d6 and d7 was decreased dramatically than that on the implantation window. The result showed that mTOR down-regulation might slow down the endometrial cell growth and proliferation, and promote endometrial cell apoptosis for the change of endometrial receptivity in response to implantation.4. The mTOR expression was in endometrial stroma cell in the different stages of early pregnant mice, which showed that mTOR was relevant with the endometrial stromal cell growth, proliferation, differentiation and apoptosis during early pregnancy.5. At the same time, the result showed that the mTOR expression in pseudopregnant group was higher slightly than that in non-pregnant group, which indicated that mTOR would be influenced limitedly on the regulation of estrogen and progestogen during the implantation process.