Screening And Expression For Differentially Expressed Genes In The Uterus Before Embryo Implantation And That Before Parturition & Effect Of IL-1β On Expression Of MMP2 In Uterus Before And After Embryo Implantation

Gestation includes a series of important stages, such as blastocyte implantation, pregnancy maintenance and parturition. In terms of immunology, uterus is a special reproductive and immune organ. It shows immunological tolerance and immunological rejection respectively at the time of blastocyte implantation and parturition; blastocyte implantation is also concerned with the action of many cytokines and MMPs. The research work can make us acquire the further recognition of molecular mechanism about blastocyte implantation and parturition and provide the theoretical basis for some leading-edge problems, such as organ transplantation, homeoclone or heteroclone. In the meantime, the result might be used as possible targets of medicine development for the cure of some reproductive diseases, such as sterility, abortion and fetal malformation. In this paper, Sprague-Dawley (SD) female rats were applied as experiment animals to screen differentially expressed genes of the uterus in different stage of pregnancy by suppression subtractive hybridization (SSH), in which the uteri from the 5^th and the 19^th day in pregnancy were respectively used as driver and tester. Seventy positive clones from forward subtractive library were selected by differential screening. Sequencing analysis and homology comparison showed that all these clones were homologous to eight known genes in GenBank by the degree from percent 90 to percent 100. These genes were differentially expressed in the uterus from the nineteenth day in pregnancy, in which the expression of uroguanylin and interferon-inducible protein 16 in the uterus in pregnancy was reported for the first time. Semi-quantitative analysis showed that the expression of uroguanylin was significantly higher in the uterus of 19th day in pregnancy than that of 5th day (P<0.001), however, there was no difference about the expression of interferon-inducible protein16. The expression of uroguanylin increased after embryo implantation, decreased on the 15th day and reached the peak on the 19th day in pregnancy. The result suggested that uroguanylin might be concerned with parturition. The expression of MMP2 was investigated by RT-PCR and immunohistochemistry, in order to study the effect of IL-1? on MMP2 in SD rat uteri before and after blastocyte implantation. The results by RT-PCR and statistical analysis showed that there was no significant difference between control group and treatment group on the 4th day and 9th day of pregnancy. On the 6th day of pregnancy the expression of MMP2 in control group was significantly higher than that in treatment group. The analysis by immunohistochemistry showed that on the 4th day of pregnancy MMP2 expressed principally at luminal epithelium and a little at matrix and gland; on the 6th day of pregnancy MMP2 mainly expressed at luminal epithelium in control group, but decreased at luminal epithelium in treatment group; on the 9th day of pregnancy MMP2 expressed at decidua. The conclusion was that IL-1? could significantly downregulate the expression of MMP2 at luminal epithelium of uterus on the 6th day of pregnancy when blastocyte implantation took place. In summary, we constructed the differentially-expressed-gene library in the uterus of parturition for the first time, in which the further research about uroguanylin suggested that it might be related to parturition. Meanwhile, we discovered that IL-1? could significantly downregulate the expression of MMP2 at luminal epithelium of uterus on the 6th day of pregnancy when blastocyte implantation took place.