Study The Impact Of PR And IL-1β、COX-2on LPS And The TLR4Antagonist-stimulated Decidual Cells

Objective:Through observing the expression of Progesterone receptor (PR) and Interleukin-1β(IL-1β),Cyclooxygenase-2(COX-2), Lipopolysaccha-ride (LPS)-stimulated decidual cells in vitro dealed with antagonists Toll-like receptor4antagonist (TLR4mAb), we explore the LPS and its antagonist’s effect on progesterone receptor of decidual cells and progesterone receptor with inflammatory cytokines’relationship.Method:Isolated and cultured human decidua of early abortion. When the cells passaged to3-4generations, HE staining and Immunofluorescence technique were used to observe the morphology and identify the purity of decidual cells. Then six pore plates were treated with the final concentration of Oug/ml,1ug/ml,3ug/ml of Toll-like receptor4monoclonal antagonist (TLR4mAb), each concentration has two pore plates. Following a24hours treatment with TLR4mAb, two does of LPS (Ong/ml and100ng/ml) were respectively added to two pore plates of every TLR4mAb concentration for24hours. Subsequently, MTT detected the survival and growth rates of cells, the levels of mRNA expression of PR, IL-1β and COX-2were detected by reverse transcription PCR (RT-PCR).Result:1. Immunofluorescence showed the3-4generation decidual cells’ volume is more increases, the nucleus is more pronounced and the purity is higher than the first generation, they have a high consistency of microscopic morphology2. MTT assay indicated that LPS inhibited growth of cells (P <0.05) compared with the control group; TLR4mAb promoted proliferation of cells, and a certain concentration of TLR4mAb can oppose growth of cell of LPS-inhibited(P>0.05), not statistically significant.3. RT-PCR(1) LPS reduced the mRNA’s expression of PR (P＜0.05), increased the mRNA’s expression of IL-1β and COX-2(P<0.05).(2) TLR4mAb was not statistically significant of the mRNA’s expression of PR and COX-2(P>0.05), reduced the mRNA’s expression of IL-1β (P＜0.05).(3) TLR4mAb increased the mRNA’s expression of PR (P<0.05) and reduced the mRNA’s expression of IL-1β (P＜0.05) after LPS-stimulated decidual cells;High concentrations of TLR4mAb reduced the mRNA’s expression of COX-2(P<0.05) after LPS-stimulated decidual cells.Conclusion:The mRNA’s expression of PR was reduced, the mRNA’s expression of IL-1β and COX-2were increased after LPS-stimulated decidual cells in vitro dealed. Toll-like receptor4antagonist antagonized the role of LPS on PR, IL-1β and COX-2.