PTPRO在妊娠期肝内胆汁淤积症孕妇胎盘组织中的表达及意义

Expression of PTPRO in placental tissues of intraheptic cholestasis of pregnancy patients and its significance

目的通过观察妊娠期肝内胆汁淤积症(ICP)与正常妊娠孕妇的胎盘来源的单核-巨噬细胞中受体O型蛋白酪氨酸磷酸酶(Protein tyrosine phosphatase receptor-type O,PTPRO)的表达情况、细胞因子的分泌及胞内信号分子NF-κB/P65活化水平的变化,探讨PTPRO在妊娠胎盘免疫微环境的平衡维持中的作用。方法选取3组孕妇:正常妊娠(26例)、ICP轻型(24例)及ICP重型孕妇(24例),分离并由密度-梯度离心法获得其胎盘来源的单核-巨噬细胞,ELISA法检测细胞培养上清液中TNF-α、IFN-γ、IL-6的水平;Real-time PCR及Western blot法分别检测PTPRO mRNA、蛋白水平的表达以及NF-κB/P65的磷酸化改变。结果与正常妊娠者相比,ICP孕妇胎盘来源的单核-巨噬细胞高分泌TNF-α、IFN-γ、IL-6(P均<0.01),NF-κB/P65的磷酸化增多,ICP轻型及重型孕妇胎盘来源的单核-巨噬细胞细胞因子PTPRO的表达上调。结论 ICP孕妇胎盘组织内炎性因子浸润,可能与PTPRO、NF-κB/P65的相互调控有关。

Objective To observe the expression of protein tyrosine phosphatase receptor-type O （PTPRO） in placenta-o- riginating monocyte-macrophage, the secretion of cytokine factors and the activation level of signal factor NF-KB/P65 in the pregnant women with intraheptic cholestasis of pregnancy（ICP） and normal pregnant women to explore the role of PTPRO in the maintaining the balance of immune microenvironment in pregnant placenta. Methods The reseach objects were divide three groups： normal pregnancy group （26 cases） , mild ICP group （24 cases）and severe ICP group （24 cases）. The placen- ta-originating monocyte-macrophages were separated and acquired by density-gradient centrifuging method. The levels of tumor necrosis factor（TNF）-ct, interferon（ IFN ）-γand interleukin （IL）-6 in the supernatant liquid of culture solution were assayed by enzyme linked immunosorbent assay（ELISA） ,and the expressions of PTPRO mRNA and protein and the phos- phorylation change of NF-KB/P65 were detected by real-time PCR and Western-blot methods,respctively. Results Com- pared with normal pregnancy group ,TNF-ct, IFN-γand IL-6 in placenta-originating monocyte-macrophages in mild ICP and severe ICP groups presented hyper-secretion levels（ all P 〈0.01 ）. The phosphorylation of NF-κB/P65 increased. The ex- pression of PTPRO in placenta-originating monocyte-macrophages in mild and severe ICP pregnant groups presented up-reg- ulation. Conclusion The invasion of inflammatory factors in placental tissues of ICP pregnant women might be related to the interaction of PTPRO and NF-κB/P65.