糖酵解相关基因在妊娠期肝内胆汁淤积症患者胎盘组织内的变化

doi:10.3877/cma.j.issn.1673-5250.2017.01.013

目的

探讨糖酵解相关基因糖转运体(GLUT)1、磷酸甘油酸激酶(PGK)1、乳酸脱氢酶(LDHA)基因在妊娠期肝内胆汁淤积症(ICP)患者胎盘组织内的表达及其意义。

方法

按照数字表法，随机选择2015年6月至10月于四川大学华西第二医院产科行择期剖宫产分娩的40例产妇的胎盘组织为研究对象。按照是否合并ICP，将其分别纳入ICP组(n＝20)和对照组(n＝20)。2组产妇行剖宫产分娩待胎盘娩出后，立即收取胎盘母体面的中央绒毛区组织。采用实时聚合酶链式反应(RT-PCR)及Western印迹法分别检测胎盘组织中GLUT1、PGK1、LDHA mRNA及蛋白相对表达水平。采用链霉菌抗生物素蛋白-过氧化物酶连结(SP)法进行免疫组化染色，并检测GLUT1、PGK1及LDHA在胎盘组织的定位。2组产妇的年龄、孕龄、孕次、产次、新生儿出生体重、娩出胎盘重量等一般临床资料比较，差异无统计学意义(P>0.05)。本研究遵循的程序符合四川大学华西第二医院人体试验委员会制定的伦理学标准，得到该委员会批准，分组征得受试对象知情同意，并与之签署临床研究知情同意书。

结果

①2组胎盘组织中GLUT1、PGK1及LDHA mRNA表达水平比较，差异均无统计学意义(P>0.05)。②ICP组胎盘组织中GLUT1、PGK1及LDHA蛋白水平均高于对照组，并且差异有统计学意义(t＝3.270，P＝0.003；t＝4.222，P<0.001；t＝3.585，P＝0.002)。③2组胎盘组织的合体滋养细胞及细胞滋养细胞的细胞质内均可见GLUT1、PGK1及LDHA蛋白的表达。

结论

ICP患者胎盘组织的糖酵解相关基因蛋白表达水平增高，糖酵解水平较正常晚孕期产妇代偿性增高，一定程度上可以缓解宫内低氧或缺氧，进一步加重的缺氧则可能导致组织能量代谢失代偿，从而导致胎死宫内等不良妊娠结局。

关键词: 妊娠, 胆汁淤积，肝内, 缺氧, 糖酵解, 孕妇

Objective

To investigate the expression of glucose transporter (GLUT1), phosphoglycerate kinase(PGK1) and lactate dehydrogenase (LDHA) in placentas of women with intrahepatic cholestasis of pregnancy (ICP).

Methods

From June to October, 2014, a total of 20 samples of placentas from women with ICP were included as the study group, and another 20 samples of normal placentas were selected as control group according to random number table. The relative expression levels of GLUT1 mRNA, PGK1 mRNA and LDHA mRNA were measured by real-time polymerase chain reaction (RT-PCR), and the relative protein levels of GLUT1, PGK1, LDHA were detected by Western blotting assay and immunohistological analysis. There were no significant differences between two groups in age, gestational age, gravidity, parity, birth weight of newborns and placental weight (P>0.05). The study protocol was approved by the Ethical Review Board of Investigation in Human Being of West China Second University Hospital, Sichuan University. Informed consent was obtained from the parents of each participating patients.

Results

① There were no statistically significant difference between two groups in the relative expression levels of GLUT1 mRNA, PGK1 mRNA and LDHA mRNA (P>0.05). ② The relative protein levels of GLUT1, PGK1 and LDHA in control group were statistically lower than those in ICP group, which with significant difference (t=3.270, P=0.003; t=4.222, P<0.001; t=3.585, P=0.002). ③ All the proteins including GLUT1, PGK1 and LDHA were detected in the cytoplasma of cytotrophoblast and syncytiotrophoblast.

Conclusions

Under hypoxic conditions in pregnant women with ICP, the increased level of glycolysis may play a role in negative pregnancy outcomes.

Key words: Pregnancy, Cholestasis, intrahepatic, Anoxia, Glycolysis, Pregnant women

表1 2组产妇胎盘组织中GLUT1、PGK1及LDHA mRNA相对表达水平比较(±s)

组别	例数	GLUT1 mRNA	PGK1 mRNA	LDHA mRNA
ICP组	20	1.7±0.6	4.8±0.5	8.2±3.1
对照组	20	1.9±0.6	4.7±0.3	6.5±3.3
t值	?	0.702	0.405	1.194
P值	?	0.491	0.690	0.248

表1 2组产妇胎盘组织中GLUT1、PGK1及LDHA mRNA相对表达水平比较(±s)

组别	例数	GLUT1 mRNA	PGK1 mRNA	LDHA mRNA
ICP组	20	1.7±0.6	4.8±0.5	8.2±3.1
对照组	20	1.9±0.6	4.7±0.3	6.5±3.3
t值	?	0.702	0.405	1.194
P值	?	0.491	0.690	0.248

表2 2组产妇胎盘组织中GLUT1、PGK1及LDHA蛋白相对表达水平比较(±s)

组别	例数	GLUT1蛋白	PGK1蛋白	LDHA蛋白
ICP组	20	1.2±0.3	1.2±0.2	1.3±0.3
对照组	20	0.9±0.2	0.8±0.2	0.8±0.4
t值	?	3.270	4.222	3.585
P值	?	0.003	<0.001	0.002

表2 2组产妇胎盘组织中GLUT1、PGK1及LDHA蛋白相对表达水平比较(±s)

组别	例数	GLUT1蛋白	PGK1蛋白	LDHA蛋白
ICP组	20	1.2±0.3	1.2±0.2	1.3±0.3
对照组	20	0.9±0.2	0.8±0.2	0.8±0.4
t值	?	3.270	4.222	3.585
P值	?	0.003	<0.001	0.002

图1 2组胎盘组织中GLUT1、PGK1及LDHA蛋白相对表达水平

图2 2组胎盘组织中GLUT1、PGK1及LDHA蛋白在细胞内的表达定位(图2A、2B：GLUT1蛋白；图2C、2D：PGK1蛋白；图2E、2F：LDHA蛋白)(SP染色，高倍镜)

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Changes of glycometabolic genes in placenta of intrahepatic cholestasis of pregnancy

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Changes of glycometabolic genes in placenta of intrahepatic cholestasis of pregnancy