摘要
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 research the role of progesterone in Toll- like receptor 4（ TLR4）- myeloid differentiation factor 88（ My D88）- dependent signaling pathway in preeclampsia patients. Methods Peripheral blood mononuclear cells（ PBMCs） from preeclampsia（ PE） patients were cultured by primary culture method,then they were treated by different concentrations of progesterone（ 0 mol/L,10^- 8mol / L,10^- 6mol / L,10^- 4mol / L）; real- time PCR was used to detect the expressions of TLR4 mRNA,My D88 mRNA,nuclear factor- κB（ NF- κB） mRNA; Western blot was used to detect the expression of IκB- α protein,gel image processing system was used to analyze grey levels of various bands; ELISA was used to detect the expressions of tumor necrosis factor（ TNF- α） and interleukin-6（ IL- 6） in liquid supernatant. Results The relative expression levels of TLR4 mRNA,My D88 mRNA,and NF- κB mRNA（ 2^- △△Ct）in different groups decreased gradually with the increase of progesterone concentration,the expression of IκB- α protein increased gradually,there were statistically significant differences（ P 〈0. 05）; meanwhile,the expressions of TNF- α and IL- 6 in liquid supernatant decreased gradually,there were statistically significant differences（ P 〈0. 05）. Conclusion Progesterone can significantly inhibit TLR4- My D88-dependent signaling pathway and protect preeclampsia patients.
出处
《中国妇幼保健》
CAS
2015年第27期4695-4698,共4页
Maternal and Child Health Care of China
