Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
Curr Med Sci. 2023 Aug;43(4):784-793. doi: 10.1007/s11596-023-2738-1. Epub 2023 Jul 5.
Gestational diabetes mellitus (GDM) is the most common metabolic disorder during pregnancy. LncRNA HLA complex group 27 (HCG27) plays a crucial role in various metabolic diseases. However, the relationship between lncRNA HCG27 and GDM is not clear. This study aimed to verify a competing endogenous RNA (ceRNA) interaction regulation axis of miR-378a-3p/mitogen-activated protein kinase 1 (MAPK1) regulated by HCG27 in GDM.
LncRNA HCG27 and miR-378a-3p were detected by RT-qPCR. The expression of MAPK1 in umbilical vein endothelial cells (HUVECs) was detected by RT-qPCR and that in the placenta by Western blotting. To explore the relationship among lncRNA HCG27, miR-378a-3p, MAPK1 and the glucose uptake ability of HUVECs, vector HCG27, si-HCG27, miR-378a-3p mimic and inhibitor were transfected to achieve overexpression and inhibition of HCG27 or miR-378a-3p. The interaction between miR-378a-3p and lncRNA HCG27 or MAPK1 was confirmed by the dual-luciferase reporter assay. Besides, glucose consumption by HUVECs was detected by the glucose assay kit.
HCG27 expression was significantly decreased in both the placenta and primary umbilical vein endothelial cells, while the expression of miR-378a-3p was significantly increased in GDM tissues, and the expression of MAPK1 was decreased in GDM tissues. This ceRNA interaction regulation axis was proved to affect the glucose uptake function of HUVECs. The transfection of si-HCG27 could significantly reduce the expression of the MAPK1 protein. If the MAPK1 overexpression plasmid was transfected simultaneously with si-HCG27 transfection, the reduced glucose uptake in HUVECs resulting from the decrease in lncRNA HCG27 was reversed. MiR-378a-3p mimic can significantly reduce the mRNA expression of MAPK1 in HUVECs, whereas miR-378a-3p inhibitor can significantly increase the mRNA expression of MAPK1. The inhibition of miR-378a-3p could restore the decreased glucose uptake of HUVECs treated with si-HCG27. Besides, overexpression of lncRNA HCG27 could restore the glucose uptake ability of the palmitic acid-induced insulin resistance model of HUVECs to normal.
LncRNA HCG27 promotes glucose uptake of HUVECs by miR-378a-3p/MAPK1 pathway, which may provide potential therapeutic targets for GDM. Besides, the fetal umbilical cord blood and umbilical vein endothelial cells collected from pregnant women with GDM after delivery could be used to detect the presence of adverse molecular markers of metabolic memory, so as to provide guidance for predicting the risk of cardiovascular diseases and health screening of offspring.
妊娠期糖尿病（GDM）是妊娠期间最常见的代谢紊乱。长链非编码 RNA HLA 复合体 27（HCG27）在各种代谢疾病中发挥关键作用。然而，lncRNA HCG27 与 GDM 之间的关系尚不清楚。本研究旨在验证 GDM 中由 HCG27 调节的 miR-378a-3p/丝裂原活化蛋白激酶 1（MAPK1）竞争性内源 RNA（ceRNA）相互作用调节轴。
通过 RT-qPCR 检测 lncRNA HCG27 和 miR-378a-3p 的表达。通过 RT-qPCR 检测脐静脉内皮细胞（HUVEC）中 MAPK1 的表达，通过 Western blot 检测胎盘组织中 MAPK1 的表达。为了探讨 lncRNA HCG27、miR-378a-3p、MAPK1 与 HUVEC 葡萄糖摄取能力之间的关系，转染载体 HCG27、si-HCG27、miR-378a-3p 模拟物和抑制剂，以实现 HCG27 或 miR-378a-3p 的过表达和抑制。通过双荧光素酶报告基因实验证实了 miR-378a-3p 与 lncRNA HCG27 或 MAPK1 之间的相互作用。此外，通过葡萄糖测定试剂盒检测 HUVEC 的葡萄糖消耗。
在 GDM 组织中，HCG27 的表达明显降低，而 miR-378a-3p 的表达明显升高，MAPK1 的表达在 GDM 组织中降低。该 ceRNA 相互作用调节轴被证明影响 HUVEC 的葡萄糖摄取功能。转染 si-HCG27 可显著降低 MAPK1 蛋白的表达。如果同时转染 si-HCG27 与 MAPK1 过表达质粒，则由于 lncRNA HCG27 的减少而导致的 HUVEC 葡萄糖摄取减少得到逆转。miR-378a-3p 模拟物可显著降低 HUVEC 中 MAPK1 的 mRNA 表达，而 miR-378a-3p 抑制剂可显著增加 MAPK1 的 mRNA 表达。抑制 miR-378a-3p 可恢复 si-HCG27 处理后 HUVEC 葡萄糖摄取的减少。此外，过表达 lncRNA HCG27 可使棕榈酸诱导的 HUVEC 胰岛素抵抗模型的葡萄糖摄取能力恢复正常。
lncRNA HCG27 通过 miR-378a-3p/MAPK1 通路促进 HUVEC 的葡萄糖摄取，这可能为 GDM 提供潜在的治疗靶点。此外，可从 GDM 孕妇产后采集胎儿脐带血和脐静脉内皮细胞，以检测代谢记忆不良分子标志物的存在，为预测心血管疾病风险和子女健康筛查提供指导。
