胎儿生长受限孕妇血清、胎盘组织中Klotho及VEGFR2蛋白的表达及其临床意义

Expression of Klotho and VEGFR2 in serum and placental tissues of pregnant women with fetal growth restriction and its clinical significance

目的探讨Klotho(KL)、VEGFR2在胎儿生长受限(fetal growth restriction,FGR)孕妇母血及胎盘组织中的表达情况。方法选取2019年1月至2020年6月于徐州医科大学附属徐州妇幼保健院首诊为FGR,并于分娩后确诊的孕妇60例,其中早发型30例(发病孕周<32周),晚发型30例(发病孕周≥32周),另随机选择同时期孕晚期正常妊娠对照组孕妇30例作为对照组。采用免疫组化SP法、Westernblot法及实时荧光定量PCR(RT-qPCR)法检测3组孕妇胎盘组织中KL、VEGFR2的定位及表达情况;ELI-SA法检测3组孕妇血清中分泌型KL(sKL)及可溶型VEGFR2(sVEGFR2)的含量。结果免疫组化法检测显示:KL蛋白主要表达于胎盘滋养细胞细胞膜、细胞质和血管内皮细胞;VEGFR2主要表达于胎盘血管内皮细胞;Western blot法及RT-qPCR法检测结果均显示:3组孕妇胎盘中KL、VEGFR2蛋白及mRNA相对表达量比较差异有统计学意义(P<0.05);FGR组表达量均低于对照组,其中早发型和晚发型比较差异无统计学意义(P>0.05);ELISA法检测显示,3组孕妇血清中sKL、sVEGFR2表达水平比较差异有统计学意义(P<0.05);FGR组表达量均低于对照组,早发型和晚发型比较差异无统计学意义(P>0.05)。结论KL、VEGFR2在孕妇体内表达水平降低可能与胎儿生长受限的发生发展有关。

Objective To explore the expression of Klotho(KL)and VEGFR2 in maternal blood and placenta of pregnant women FGR.Methods A total of 60 pregnant women with fetal growth restriction who were first diagnosed in Xuzhou Maternal and Child Health Hospital affiliated to Xuzhou Medical University from January2019 to June 2020 and diagnosed after delivery were selected,including 30 in the early-onset group(gestational week of onset<32 weeks),30 cases in the late onset group(gestational week of onset≥32 weeks),and 30 cases of normal full-term pregnant women were randomly selected as the control group.The localization and expression of KL and VEGFR2 in placental tissues of 3 groups of pregnant women were detected by immunohistochemistry(IHC),Western blot and RT-qPCR.ELISA was used to detect the sKL and sVEGFR2 in serum of the three groups of pregnant women.Results IHC showed that KL protein was mainly expressed in the cell membrane of placental trophoblast cells and vascular endothelial cells,VEGFR2 is mainly expressed in placental vascular endothelial cells.Western blot and RT-qPCR detection results showed that there were statistically significant differences in the relative expression levels of KL and VEGFR2 between the three groups(P<0.05).The expression level of FGR groups was lower than that of the control group,and there was no statistical difference between the early-onset group and late-onset group(P>0.05).ELISA showed that the serum levels of sKL and sVEGFR2 in the three groups were statistically different(P<0.05).The expression level of FGR group was lower than that of the control group,and there was no statistical difference between the early-onset group and the late-onset group(P>0.05).Conclusion The decreased expression of KL and VEGFR2 in pregnant women may be related to the occurrence and development of fetal growth restriction.