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目的分析β-链蛋白(β-catenin)及重组人Dickkopf相关蛋白1(DDK1)在不明复发性流产(unexplained recurrent spontaneous abortion,URSA)患者的绒毛滋养层细胞和蜕膜组织中表达,为URSA的诊断与治疗提供理论依据。方法以2014年2月~2016年2月,医院收治的URSA患者40例,作为研究对象,入选病例组,另选择同期收治的正常妊娠早期女性40例,纳入对照组。人工流产术后,取蜕膜以及绒毛组织,一部分迅速放入无RNA酶的EP管中,液氮下保存,进行实时定量PCR,免疫组化染色镜下检查,另一部分直接采用4%福尔马林溶液固定,进行蛋白提取与Western blot分析。结果病例组的绒毛组织、蜕膜组织β-catenin蛋白光密度与细胞中β-catenin蛋白表达低于对照组,差异有统计学意义(P<0.05);病例组绒毛组织、蜕膜组织β-catenin蛋白高表达率低于对照组,病例组细胞滋养细胞膜浅黄色着色率低于对照组,差异有统计学意义(P<0.05)。病例组绒毛组织、蜕膜组织中的DKK1 m RNA水平分别是对照组的1.84、2.34倍。结论 URSA患者的绒毛滋养层细胞和蜕膜组织中β-catenin蛋白表达显著下降,DDK1 m RNA水平上升。
Objective To investigate the expression of β-catenin and recombinant human Dickkopf-related protein 1 (DDK1) in villus trophoblast cells and decidual tissue in patients with unexplained recurrent spontaneous abortion (URSA) Diagnosis and treatment provide a theoretical basis. Methods From February 2014 to February 2016, 40 hospitalized patients with URSA were enrolled in the study. Forty patients with normal early pregnancy who were treated at the same period were enrolled in the control group. After artificial abortion, the decidua and villus tissues were taken out, and some of them were rapidly put into RNase-free EP tube and stored in liquid nitrogen for real-time quantitative PCR and immunohistochemical examination under the microscope. The other part was directly treated with 4% Marlin solution fixation, protein extraction and Western blot analysis. Results The optical density of β-catenin and the expression of β-catenin protein in the chorionic and decidual tissues were lower than those in the control group (P <0.05). In the cases, the expression of β-catenin in the chorionic and decidual tissues, The high expression rate of catenin was lower than that of the control group. The staining rate of buffy coat cells in the case group was lower than that of the control group (P <0.05). In the case group, the level of DKK1 m RNA in the chorionic and decidual tissues was 1.84 and 2.34 times that of the control group respectively. Conclusions The expression of β-catenin in villus trophoblast cells and deciduas of URSA patients is significantly decreased, and the level of DDK1 m RNA is increased.