Obstetrics and Gynecology Department, University of Hong Kong Shenzhen Hospital, 518000, Shenzhen, Guangdong, China.
Obstetrics and Gynecology Department, Nanfang Hospital Affiliated to Southern Medical University, 510515, Guangzhou, Guangdong, China.
Placenta. 2021 Feb;105:14-22. doi: 10.1016/j.placenta.2021.01.015. Epub 2021 Jan 19.
Dysregulated genes in glucose transport and metabolize pathways have been found in patients with Gestational diabetes (GDM), but the underlying mechanisms were still unclear.
Placental villous samples were collected from 31 patients with GDM and 20 healthy controls. The expression of GLUT1, GLUT4, GLUT9 and HK2 was examined by immunoblotting and qRT-PCR. The miRNAs have the potential targeting GLUT1 and HK2 were predicted using online bioinformatics tool: TargetScan. The interaction between miRNAs and target genes were confirmed by dual luciferase assay and immunoblotting. The function of miR-9 and miR-22 on glucose metabolism was examined by glucose uptake assay and lactate secretion assay.
GLUT1 and HK2 proteins level was found upregulated in patients with GDM, but the mRNA level was not significantly changed. Predicted by using bioinformatics tools and confirmed by dual luciferase assay and immunoblotting, GLUT1 was identified as a target of miR-9 and miR-22, whereas HK2 was identified as a target of miR-9. MiR-9 and miR-22 level was found reduced in the placenta villous and negatively correlated with the expression of GLUT1 and HK2. Functional studies indicated that miR-9 and miR-22 inhibitors upregulated the expression of GLUT1 and HK2, and then increased the glucose uptake, lactate secretion, cell viability and repressed apoptosis in primary syncytiotrophoblasts (STBs) and HTR8/SVneo cells.
The upregulation of GLUT1 and HK2 in the placenta, which is induced by miR-9 and miR-22 reduction, contributes to the disordered glucose metabolism in patients with GDM.
患有妊娠糖尿病（GDM）的患者中发现葡萄糖转运和代谢途径的失调基因，但潜在机制尚不清楚。
从 31 名 GDM 患者和 20 名健康对照者中收集胎盘绒毛样本。通过免疫印迹和 qRT-PCR 检测 GLUT1、GLUT4、GLUT9 和 HK2 的表达。使用在线生物信息学工具 TargetScan 预测可能靶向 GLUT1 和 HK2 的 miRNA。通过双荧光素酶测定和免疫印迹证实 miRNA 与靶基因之间的相互作用。通过葡萄糖摄取测定和乳酸分泌测定检查 miR-9 和 miR-22 对葡萄糖代谢的功能。
发现 GDM 患者的 GLUT1 和 HK2 蛋白水平升高，但 mRNA 水平没有显著变化。通过生物信息学工具预测并通过双荧光素酶测定和免疫印迹证实，GLUT1 被鉴定为 miR-9 和 miR-22 的靶标，而 HK2 被鉴定为 miR-9 的靶标。miR-9 和 miR-22 水平在胎盘绒毛中发现降低，与 GLUT1 和 HK2 的表达呈负相关。功能研究表明，miR-9 和 miR-22 抑制剂上调 GLUT1 和 HK2 的表达，然后增加原代合体滋养细胞（STB）和 HTR8/SVneo 细胞中的葡萄糖摄取、乳酸分泌、细胞活力并抑制细胞凋亡。
miR-9 和 miR-22 减少诱导胎盘 GLUT1 和 HK2 的上调，导致 GDM 患者葡萄糖代谢紊乱。
