Effect Of Brucella LPS On Endometrium Decidualization Of Mouse

Brucella lipopolysaccharide(LPS), as the main ingredient of brucella outer membrane, is one of the virulence factors of brucella, potentially toxic, and has been widely used as a model to explore mechanism of pregnancy loss induced by LPS in mice. Endometrial decidual, a kind of special process of differentiation, endometrial in normal embryo pregnancy and implantation, plays an important role embryo implantation. In order to study the relations between brucella lipopolysaccharide and pregnancy mice endometrial decidual, multiple techniques were performed, such as histologic observation, real-time quantitative PCR, isolation, culture stromal cells, artificial induction decidua and western blotting. The present study mainly aim to explore the influence of Brucella LPS in pregnant mice endometrial decidual process and the mechanism of Brucella LPS in the early abortion, which will provide a basis to explore prevention, control and testing new targets of Brucella. The results are as follows:1. Brucella LPS were used as drugs to establish pregnant mice abortion model. Pregnancy mice were divided into 4 groups, including 0(control group) 2, 5, 10 10μg/g doses of LPS group, each group of 6 to 8 pregnant mice, only in pregnancy by intraperitoneal injection LPS at 4d, pregnancy of 6d sample were observed. Results showed that LPS induced embryonic implantation failure in a dose dependent manner. When LPS dose reached to10μg/g, implantation of mouse embryos rate was 0%. While 5μg/g LPS group do not have good effect, so 5μg/g LPS is suitable for using to create embryonic implantation failure model.2. HE staining observation found fibrinolysis appear in decidual area at pregnant 6d; during the 8 d, near the embryo of decidual cells are large granular cell infiltration, decidual area has a large gap, embryonic cell necrosis occur compared with control group. All these indicated that brucella LPS make early gravid uterus tissue morphological changes. Real-time quantitative PCR and Western Blot were used to study the effects of Brucella LPS on endometrial decidual related factors(IGFBP-1 and CyclinD3) change in the pregnancy mice. Results show that the Brucella LPS in pregnant 5-8 d mouse endometrial decidual is changed in the process of pregnancy. In addition, LPS inhibited endometrial decidual mark factors expression levels, which suggested brucella LPS may adjust the mouse endometrial decidual process, one of the reasons may lead to pregnancy failure in mice.3. In order to study effect of Brucella LPS on decidual process in virtro, Brucella LPS was added to endometrial stromal cells that was cultured and inducted decidua. Real-time fluorescent quantitative PCR were used to detect brucella LPS on the effects of the decidual related factors such as CyclinD3 and dPRP expression. Results showed that the brucella LPS reduce the decidual mark molecular expression, which indicate Brucella LPS can disrupt the endometrial decidual process, influence the normal pregnancy of mice.