The Study Of Progesterone Receptor(PR) And Tumor Nuclear Factor-alpha(TNF-α) Express In Human Decidual Cell Which Effected By The Lipopolysacchride(LPS)

Object:To study of the effects of lipopolysaccharide on the expression of progesterone receptor (PR) and TNF-a in human decidual cell cultured in vitro, and to explore the mechanism of spontaneous abortion caused by bacterial infection.Methods:Decidual cells in primary culture were isolated from term placentas of 10 normal early pregnant women undergoing induced abortion, and were identified by the method of immunocytochemistry of prolactin. In this study, the 4th to 5th passages of human decidual cells were used and randomly divided into a control group and four experiment groups. In all groups, the cells were cultured in 20% FBS/DMEM/F12 medium. In addition, the cells of four experiment groups were treated with the final concentration of 25ng/ml,50ng/ml, 100ng/ml, and 200ng/ml of lipopolysaccharide for 24h, respectively. The expression intensity of PR and TNF-a were detected by laser confocal microscopy after immnofluorescence staining with Cy3, and the data were analyzed using Image J software. The database was established and analyzed using SPSS 17.0 software package. The one-way ANOVA was used to determine the difference between means of the groups. The linear regression analysis was used to analyze the association between the expression of PR and that of TNF-α.Results:1. Human decidual cells were successfully cultured in vitro with high survival rate,95.0% of which are positive immunocy- tochemical staining for prolactin.2. Compared with the control group, the experimental group showed statistical significantly decrease in the average fluorescence intensity of PR (P< 0.01). Moreover, the fluorescence intensity of PR is decreased with the increasing concentration of lipopolysaccharide.3. Compared with the control group, the experimental group showed statistical significantly increase in the average fluorescence intensity of TNF-a (P< 0.01). Moreover, the fluorescence intensity of TNF-a is increased with the increasing concentration of lipopolysaccharide.4. In vitro culture, the fluorescence intensities of PR in decidual cells were negatively correlated with those of TNF-a (r=-0.841,P<0.01)Conclusions:1. Lipopolysaccharide inhibits the expression of progesterone receptor in human decidual cells. The higher concentration of lipopolysaccharide is in the culture, the lower PR is expressed.2. Lipopolysaccharide enhances the expression of TNF-αin human decidual cells. The higher concentration of lipopolysaccharide is in the culture, the more TNF-a is expressed.3. In vitro culture, the expression of PR in human decidual cells is negatively correlated with that of TNF-α.