Expression of TLR4 and its effect on Treg cells in early pregnancy decidual stromal cells after lipopolysaccharide treating

• Published in: European journal of obstetrics & gynecology and reproductive biology 2019-06, Vol.237, p.209-214
• Main Authors: Wu, Ali, Liu, Fumin, Liu, Xiaoyun, Feng, Xia, Lin, Hua
• Format: Article
• Language: English
• Subjects: Adult; Cell Differentiation - drug effects; Decidua - drug effects; Decidua - metabolism; Decidual stromal cells; Female; Forkhead Transcription Factors - metabolism; Humans; Lipopolysaccharides - pharmacology; Maternal-fetal immunity; NF-kappa B - metabolism; NF-KappaB Inhibitor alpha - metabolism; Pregnancy; Regulatory T cells; Signal Transduction - drug effects; Stromal Cells - drug effects; Stromal Cells - metabolism; T-Lymphocytes, Regulatory - drug effects; T-Lymphocytes, Regulatory - metabolism; Toll-like receptor 4; Toll-Like Receptor 4 - metabolism; Young Adult
• ISSN: 0301-2115; 1872-7654
• DOI: 10.1016/j.ejogrb.2018.12.006

To investigate the expression of TLR4 in human early pregnancy decidual stromal cells (DSCs) induced by lipopolysaccharide (LPS) and its effect on the peripheral blood regulatory T (Treg) cells subgroup in women of childbearing age. Isolating and cultivating normal human early pregnancy DSCs followed by treatment with 0, 25, 50, 100 and 200 ng/ml LPS, and the expression level of TLR4 mRNA was detected by RT-PCR. After 3 or 4 generation we divide the DSCs into 5 groups: ①Control group: Cultivation of peripheral blood lymphocyte (PBLC); ②Co-cultivation group: Co-cultivation of PBLC and DSCs; ③LPS stimulation group: PBLC + DSCs + LPS; ④PDTC blocking-up group: PBLC + DSCs + LPS + PDTC; ⑤TLR4 blocking-up group: PBLC + DSCs + LPS + TLR4mAb. In ①–④ groups, western blot was used to detect the expression of inhibitory factor-κB (IκB-α) protein and RT-PCR was used to detect the expression of FoxP3 mRNA. In ①–⑤ groups, flow cytometry was applied to detect the percentage of Treg cells subgroup. The purity of primary cultured DSCs was more than 95%. RT-PCR results showed that the expression level of TLR4 mRNA increased gradually with the augment of LPS concentration. Western blot and RT-PCR showed that the expression of IκBα protein and FoxP3 mRNA in the other 3 groups was significantly higher than that in the control group (P