2019 年第 12 期 第 14 卷

微小RNA-132-3p对妊娠期糖尿病患者胎盘线粒体功能和葡萄糖代谢的影响

Influence of microRNA-132-3p on mitochondrial function and glucose metabolism of placenta in patients with gestational diabetes mellitus

作者：赵骏达郭春凤马俊旗

英文作者：

单位：830011乌鲁木齐，新疆医科大学第一附属医院妇科中心

英文单位：

关键词：妊娠期糖尿病；微小RNA-132-3p；葡萄糖转运蛋白1；己糖激酶2

英文关键词：

• 摘要：

• 【摘要】目的    探讨微小RNA-132-3p(miR-132-3p)对妊娠期糖尿病患者胎盘线粒体功能和葡萄糖代谢的影响。方法    选取2016年1月至2018年9月于新疆医科大学第一附属医院妇科中心行剖宫产的妊娠期糖尿病患者20例，按照治疗方法分为饮食控制组（10例，单独使用饮食治疗）和药物组（10例，使用格列本脲或胰岛素治疗），收集患者剖宫产后胎盘组织。另选取同期于本院住院的无妊娠并发症的健康女性10名为对照组，收集分娩后胎盘组织。采用酶联免疫吸附试验法检测3组入选者胎盘匀浆中人胎盘催乳素（hPL）含量，以逆转录-聚合酶链反应（RT-PCR）法检测胎盘糖代谢相关指标葡萄糖转运蛋白1（GLUT1）、己糖激酶2（HK-2）、磷酸果糖激酶（PFK）、乳酸脱氢酶（LDH）和线粒体呼吸功能相关指标过氧化物酶体增殖活化受体γ辅助活化因子1α（PGC-1α）、过氧化物酶体增殖物受体γ（PPARγ）表达，采用TaqMan法测定miR-132-3p的表达，分离培养绒毛滋养细胞，以荧光素酶报告基因法寻找miR-132-3p靶基因，以miR-132-3p mimcs转染细胞，RT-PCR检测GLUT1、HK-2、PPARγ表达改变。结果    药物组患者年龄、胎盘质量明显高于对照组［(35.3±1.4)岁比(26.1±3.2)岁、(834±50)g比(693±40)g］，胎儿/胎盘质量比明显低于对照组［(3.444±0.023)比(5.271±0.021)］，差异均有统计学意义（均P＜0.05）。饮食控制组hPL含量明显高于对照组，药物组hPL含量明显高于饮食控制组和对照组［(28.6±1.0)mg/mg蛋白比(20.3±1.2)、(10.4±0.7)mg/mg蛋白］，差异均有统计学意义（均P＜0.05）。与对照组和饮食控制组比较，药物组胎盘中糖代谢相关指标GLUT1、HK-2、PFK和LDH显著上调，与对照组比较，饮食控制组、药物组中线粒体呼吸功能相关指标PGC-1α、PPARγ显著下调，且药物组PGC-1α、PPARγ显著低于饮食控制组，差异均有统计学意义（均P＜0.05）。miR-132-3p在药物组胎盘组织及绒毛滋养细胞中相对表达量均明显低于对照组和饮食控制组［（0.54±0.20）比（1.01±0.12）、（1.11±0.24），（0.61±0.19）比（1.03±0.09）、（1.14±0.20）］，差异均有统计学意义（均P＜0.05）。miR-132-3p可靶向调控GLUT1表达。以miR-132-3p mimics转染药物组绒毛滋养细胞后，GLUT1和HK-2表达明显降低，而PPARγ表达明显升高，差异均有统计学意义（均P＜0.05）。结论    妊娠期糖尿病患者hPL明显升高，线粒体功能降低，糖酵解增加，miR-132-3p表达降低，miR-132-3p表达可调控胎盘绒毛滋养细胞糖酵解。

• 【Abstract】Objective    To explore the influence of microRNA-132-3p(miR-132-3p) on mitochondrial function and glucose metabolism of placenta in patients with gestational diabetes mellitus. Methods    Twenty patients with gestational diabetes mellitus undergoing cesarean in the First Affiliated Hospital of Xinjiang Medical University were enrolled from January 2016 to September 2018. According to different therapeutic methods, they were divided into simple diet control group(n=10) and medication group(n=10, treated with glibenclamide or insulin). Ten healthy women with normal labor were enrolled as control group. Placenta tissues were separated after delivery. Human prolactin(hPL) content in placenta homogenate was detected by enzyme-linked immunosorbent assay. Glucose metabolism indicators including glucose transporter-1(GLUT1), hexokinase-2(HK-2), phosphofructokinase(PFK) and lactic dehydrogenase(LDH), and mitochondrial respiratory function indexes including peroxisome proliferator-activated receptor-γ coactivator-1α(PGC-1α) and peroxisome proliferator-activated receptor-γ(PPARγ) were detected by reverse transcription polymerase chain reaction（RT-PCR）. Expression of miR-132-3p was detected by TaqMan method. Target gene of miR-132-3p was identified by luciferase assay. Placenta chorionic trophoblasts were isolated, cultured and transfected by miR-132-3p mimcs; changes in GLUT1, HK-2 and PPARγ expression were detected by RT-PCR. Results    Age and placenta mass in the medication group were higher［(35.3±1.4)years vs (26.1±3.2)years, (834±50)g vs (693±40)g］ and the fetus to placenta weight ratio was lower［(3.444±0.023) vs (5.271±0.021)］ compared with those in the control group(all P＜0.05). The content of hPL in the diet control group was higher than that in the control group, and it was higher in the medication group than that in the diet control group and control group［(28.6±1.0)mg/mg vs (20.3±1.2),(10.4±0.7)mg/mg］（all P＜0.05）. In the medication group there were higher expressions of GLUT1, HK-2, PFK and LDH than the control group and diet control group（all P＜0.05）. In the diet control group and medication group there were lower expressions of PGC-1α and PPARγ; the expression levels in the medication group were lower than those in the diet control group（all P＜0.05）. Expressions of miR-132-3p in placenta and chorionic trophoblasts in the medication group were lower than those in the control group and diet control group［（0.54±0.20） vs （1.01±0.12）,（1.11±0.24）; （0.61±0.19） vs （1.03±0.09）,（1.14±0.20）］(all P＜0.05). MiR-132-3p showed target regulatory effect on the expression of GLUT1. After transfection with miR-132-3p mimics, expressions of GLUT1 and HK-2 significantly decreased while PPARγ expression significantly increased in chorionic trophoblasts(P＜0.05). Conclusions    In patients with gestational diabetes mellitus, hPL content increases, mitochondrial function reduces, glycolysis increases and miR-132-3p expression decreases in the placenta. The expression of miR-132-3p can regulate the glycolysis of placental chorionic trophoblast cells.