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目的研究孕激素在子痫前期(preeclampsia,PE)孕妇Toll样受体4(toll-like receptor4,TLR4)-髓样细胞分化蛋白88(myeloid differentiation factor 88,My D88)依赖的信号通路中的作用。方法原代培养子痫前期孕妇外周血单个核细胞(peripheral blood mononuclear cell,PBMC),以不同浓度孕激素(0 mol/L、10-8mol/L、10-6mol/L、10-4mol/L)处理,实时定量聚合酶联反应(Real-time polymerase Chain reaction,real-time PCR)检测TLR4 mRNA、My D88 mRNA、核转录因子-κB(nuclear factorκB,NF-κB)mRNA的表达;免疫印迹法Western blot检测IκB-α蛋白的表达,用凝胶图像处理系统分析各条目标带灰度值酶联免疫吸附试验(Enzyme-linked immunosorbent assay,ELISA)检测细胞上清液中肿瘤坏死因子-α(tumor necrosis factors,TNF-α)及白介素-6(interleukin-6,IL-6)的表达。结果随着孕酮浓度的增加,各组TLR4 mRNA、My D88mRNA及NF-κB mRNA的相对表达量2-△△Ct均逐渐降低,IκB-α蛋白的表达逐渐增加,差异均有统计学意义(均P<0.05);同时,细胞上清液中细胞因子TNF-α及IL-6的表达均逐渐减少,差异均有统计学意义(P<0.05)。结论孕激素可显著抑制TLR4-My D88依赖的信号传导通路,对子痫前期患者具有保护作用。
Objective To study the role of progesterone in toll-like receptor 4 (TLR4) -myeloid differentiation factor 88 (My D88) -dependent signaling pathway in preeclampsia (PE) . Methods Peripheral blood mononuclear cells (PBMCs) of pregnant women with preeclampsia were cultured in primary culture with different concentrations of progesterone (0 mol / L, 10-8 mol / L, 10-6 mol / L, 10-4 mol / L ), And the expression of TLR4 mRNA, My D88 mRNA and NF-κB mRNA were detected by real-time polymerase chain reaction (Real-time PCR) Western blot was used to detect the expression of IκB-α protein. Gel imaging system was used to analyze the gray value of each target with enzyme-linked immunosorbent assay (ELISA) to detect the levels of tumor necrosis factor-α tumor necrosis factors (TNF-α) and interleukin-6 (IL-6). Results With the increase of progesterone concentration, the relative expression of TLR4 mRNA, My D88 mRNA and NF-κB mRNA in each group decreased gradually and the expression of IκB-α protein gradually increased (P <0.05). The difference was statistically significant All P <0.05). Meanwhile, the expression of cytokines TNF-α and IL-6 in the supernatant of the cells decreased gradually, the differences were statistically significant (P <0.05). Conclusions Progesterone can significantly inhibit TLR4-My D88-dependent signaling pathway and have protective effects on preeclampsia patients.