缺氧诱导因子-1α诱导子痫前期胎盘滋养细胞中GLUT3上调的机制研究  

Mechanism of hypoxia-inducible factor-1α induced GLUT3 upregulation in placental trophoblast cells of preeclampsia

作　　者：邢雪凤 吴小颖 廖鹏飞 曾浩岚 安明 XING Xuefeng;WU Xiaoying;LIAO Pengfei;ZENG Haolan;AN Ming(Department of Obstetrics and Gynecology,Sanya People’s Hospital,Sanya,Hainan 572000,China)

机构地区：[1]三亚市人民医院妇产科,海南三亚572000

出　　处：《中国优生与遗传杂志》2023年第3期533-538,共6页Chinese Journal of Birth Health & Heredity

基　　金：海南省卫生健康行业科研项目(19A200151)。

摘　　要：目的在子痫前期(PE)患者胎盘组织中探究葡萄糖转运蛋白3(GLUT3)的表达及其可能的调控机制。方法RT-PCR与Western blot法检测PE患者(PE组)和健康妊娠女性(对照组)的胎盘组织中GLUT3与缺氧诱导因子-1α(HIF-1α)的mRNA和蛋白表达差异;分别使用1%、5%与20%浓度的氧进行干预人绒毛膜滋养细胞HTR8/SVneo,Westernblot法检测上述不同浓度氧干预下的细胞中GLUT3与HIF-1α表达;将沉默HIF-1α的siRNA(si-HIF-1α)和阴性对照序列(si-NC)转染入细胞中,并采用1%、5%和20%浓度的氧进行干预转染后的细胞,Western blot检测不同氧浓度下细胞中GLUT3蛋白的表达;将携带缺氧反应元件HRE野生型(HRE-WT)和突变型(HRE-MUT)的Pmir-REPORTTM荧光素酶载体质粒与si-HIF-1α、si-NC共转染入细胞中,荧光素酶基因报告实验检测1%和20%浓度氧干预下的细胞中荧光素酶活性。结果与对照组相比,PE组胎盘组织中GLUT3与HIF-1α的mRNA和蛋白表达均明显升高;与20%浓度的氧相比,5%与1%浓度的氧处理的HTR8/SVneo细胞中HIF-1α与GLUT3的蛋白表达均明显降低;与5%或1%氧浓度干预的si-NC组相比,si-HIF-1α组中滋养细胞中GLUT3蛋白在20%、5%和1%浓度氧处理下的表达均明显降低;与20%浓度氧条件下的HRE-WT+si-NC组相比,1%浓度氧条件下的HRE-WT+si-NC组细胞中荧光素酶强度明显降低,而20%与1%浓度氧条件下的HRE-WT+si-HIF-1α组、HRE-MUT+si-NC组和HRE-MUT+si-HIF-1α组细胞中荧光素酶活性均无明显改变。结论GLUT3在PE胎盘组织的表达明显升高,这可能与低氧环境能通过HIF-1α在转录水平上促进滋养细胞中GLUT3的表达有关。Objective To investigate the expression of glucose transporter 3(GLUT3)in placenta of patients with preeclampsia(PE)and its possible regulatory mechanism.Methods The mRNA and protein expressions of GLUT3 and hypoxia-inducible factor-1α(HIF-1α)in placenta of PE patients(PE group)and healthy pregnant women(control group)were detected by RT-PCR and Western blot method.Human chorionic trophoblast cells HTR8/SVneo were treated with 1%,5% and 20% oxygen,respectively.Western blot was used to detect the expression of GLUT3 and HIF-1α in the cells treated with different oxygen concentrations.HIF-1α silencing siRNA(si-HIF-1α)and negative control sequence(si-NC)were transfected into cells,and 1%,5% and 20% oxygen concentrations were used to intervene the transfected cells.Western blot was used to detect the expression of GLUT3 protein in the cells under different oxygen concentrations.The luciferase vector plasmid Pmir-REPORTTM carrying HRE wild-type(HRE-WT)and mutant(HRE-MUT)of hypoxia reaction elements was co-transfected into cells with si-HIF-1α group and si-NC group.Luciferase gene reporting assay was used to detect luciferase activity in cells in various group.Results Compared with control group,the expression levels of GLUT3 and HIF-1αin placental tissue of PE group were significantly increased.Compared with 20% oxygen,the protein expression levels of HIF-1α and GLUT3 in HTR8/SVneo cells treated with 5% and 1%oxygen were significantly decreased.Compared with si-NC group treated with 5% or 1% oxygen concentration,the protein expression level of GLUT3 in trophoblast cells in si-HIF-1αgroup were significantly decreased under 20%,5% and 1% oxygen concentration.Compared with HRE-WT+si-NC group at 20%oxygen concentration,the luciferase intensity of cells in HRE-WT+si-NC group at 1% oxygen concentration was significantly decreased.However,the luciferase activity in HRE-WT+si-HIF-1αgroup,HRE-MUT+si-NC group and HRE-MUT+si-HIF-1α group under 20% and 1% oxygen concentration had no significant change.Conclusion GLUT3

关 键 词：子痫前期 缺氧诱导因子-1Α 葡萄糖转运蛋白3 滋养细胞 缺氧反应元件HRE 

分 类 号：R714.244[医药卫生—妇产科学]