巨大儿胎盘组织中Klotho mRNA和蛋白的表达及其与新生儿出生体质量的关系

摘要

目的：探讨巨大儿胎盘组织中Klotho mRNA和蛋白的表达与其出生体质量的关系。方法将2014年11月至2015年3月在中国医科大学附属盛京医院分娩的足月孕妇120例，根据孕妇是否合并GDM（除GDM外均无其他妊娠期并发症及合并症）及其新生儿出生体质量分为4组：GDM巨大儿组（孕妇患GDM，其新生儿为巨大儿），GDM正常出生体质量儿组（孕妇患GDM，其新生儿出生体质量正常）；无GDM巨大儿组（健康孕妇，其新生儿为巨大儿），无GDM正常出生体质量儿组（健康孕妇，其新生儿出生体质量正常）。采用免疫组化SP法、实时荧光定量PCR技术及蛋白印迹法检测胎盘组织中Klotho mRNA和蛋白的表达，分别比较各组表达水平的差异。结果（1）免疫组化SP法检测显示，GDM巨大儿组中Klotho蛋白阳性表达率（93%，28/30）较GDM正常出生体质量儿组（73%，22/30）明显增加（P<0.05）；无GDM巨大儿组中Klotho蛋白阳性表达率（97%，29/30）较无GDM正常出生体质量儿组（80%，24/30）明显增加（P<0.05）。（2）实时荧光定量PCR技术检测显示，GDM巨大儿组Klotho mRNA表达水平（4.3±3.1）较GDM正常出生体质量儿组（2.1±2.4）明显增加（P<0.05）；无GDM巨大儿组Klotho mRNA表达水平（4.8±3.4）较无GDM正常出生体质量儿组（2.6±3.3）明显增加（P<0.05）。（3）蛋白印迹法检测显示，GDM巨大儿组Klotho蛋白的表达水平（1.27±0.90）较GDM正常出生体质量儿组（0.64±0.24）明显增加（P<0.05）；无GDM巨大儿组Klotho蛋白的表达水平（2.51±3.52）较无GDM正常出生体质量儿组（0.77±0.37）明显增加（P<0.05）。（4）3种方法均提示GDM巨大儿组较无GDM巨大儿组、GDM正常出生体质量儿组较无GDM正常出生体质量儿组Klotho mRNA和蛋白表达水平无明显差异（P>0.05）。结论 Klotho基因在胎盘组织中表达上调可能是导致巨大儿发生的机制之一，Klotho基因对于新生儿出生体质量的调节与是否合并GDM无明显关联。%Objective To explore the the expression of Klotho mRNA and protein in placenta of macrosomia and its relationship with the birth weight of neonates. Methods The cases were from November 2014 to March 2015 in Shengjing Hospital of China Medical University, divided into 4 groups:the gestational diabetes with macrosomia group (GM), the gestational diabetes with normal birth weight group (GN), the normal pregnancy with macrosomia group (NM) and the normal pregnancy with normal birth weight group (NN). Klotho mRNA and protein expression in the placenta were detected by immunohistochemistry SP method, real-time fluorescent quantitative PCR and western blot, respectively, and were compared among the 4 groups. Results (1) Immunohistochemical detection showed the positive rate of Klotho protein was significantly higher in the placenta of GM (93%,28/30) than in the GN (73%,22/30;P<0.05). The positive rate was significantly higher in the placenta of NM (97%,29/30) than in the NN (80%,24/30;P<0.05). (2) Real-time fluorescent quantitative PCR showed the Klotho mRNA expression was significantly higher in the placenta of GM (4.3 ± 3.1) than in the GN (2.1 ± 2.4;P<0.05). The Klotho mRNA expression was also significantly higher in the placenta of NM (4.8 ± 3.4) than in the NN (2.6 ± 3.3;P<0.05). (3) Western blot showed the Klotho protein expression was significantly higher in the placenta of GM (1.27±0.90) than in the GN (0.64±0.24;P<0.05). It was also significantly higher in the placenta of NM (2.51±3.52) than in the NN (0.77±0.37;P<0.05). (4) There were no significant differences in the expression of Klotho mRNA and protein between GM and NM, GN and NN (P>0.05). Conclusions The up-regulation of Klotho gene may be associated with macrosomia. The relationship is not affected by the complication of gestational diabetes.