Department of Endocrinology and Metabolism, Nanfang Hospital, Southern Medical University , Guangzhou, Guangdong Province, China.
Department of Internal Medicine, South Branch of Fujian Provincial Hospital , Fuzhou, Fujian Province, China.
Cell Cycle. 2021 Feb;20(4):353-368. doi: 10.1080/15384101.2020.1870335. Epub 2021 Jan 28.
Insulin resistance (IR) is a complex pathological condition resulting from the dysregulation of cellular response to insulin hormone in insulin-dependent cells and is recognized as a pathogenic hallmark and strong risk factor for metabolic syndrome. The present study aims to elucidate the molecular mechanism of the pathogenesis of IR. Here, we used human umbilical vein endothelial cells (HUVECs) to establish the IR cell model induced by 1 × 10 mmol/L insulin. After 48 h, reactive oxygen species (ROS) and glucose consumption were measured by DCFH-DA and GOD-POD methods, respectively. The results of Microarray analysis demonstrated that there were 10 differentially expressed miRNAs (DEMs) selected based on Fold change (FC) and P value in the IR cell model compared with HUVECs. The enriched gene ontology (GO) terms analysis showed that the target genes of these 10 DEMs were significantly enriched in biological process, cellular component and molecular function, and the significantly enriched Kyoto Encyclopedia of Genes or Genomes (KEGG) pathways mainly include AMPK signaling pathway and PI3K signaling pathway. Amongst all, the expression level of miR-3138 was highest in the IR cell model evaluated by qRT-PCR. Through Targetscan, KSR2 mRNA was predicted as a target of miR-3138. And mRNA and protein expression levels of miR-3138, KSR2, GLUT4, AMPK, PI3K, Akt were examined using qRT-PCR and Western blotting, respectively. The interaction between miR-3138 and KSR2 was evaluated by dual-luciferase reporter assay. Our results showed that miR-3138 significantly deteriorated the IR of HUVECs via KSR2/AMPK/GLUT4 signaling pathway.
胰岛素抵抗（IR）是一种复杂的病理状态，源于胰岛素依赖性细胞中胰岛素激素细胞反应的失调，被认为是代谢综合征的发病标志和强危险因素。本研究旨在阐明 IR 发病机制的分子机制。在这里，我们使用人脐静脉内皮细胞（HUVEC）建立了由 1×10 mmol/L 胰岛素诱导的 IR 细胞模型。48 小时后，通过 DCFH-DA 和 GOD-POD 方法分别测量活性氧（ROS）和葡萄糖消耗。微阵列分析的结果表明，与 HUVEC 相比，IR 细胞模型中选择了 10 个具有差异表达的 miRNA（DEM），基于倍数变化（FC）和 P 值。富集基因本体论（GO）术语分析表明，这些 10 个 DEM 的靶基因在生物学过程、细胞成分和分子功能中显著富集，显著富集的京都基因与基因组百科全书（KEGG）途径主要包括 AMPK 信号通路和 PI3K 信号通路。在所有这些 miRNA 中，qRT-PCR 评估的 IR 细胞模型中 miR-3138 的表达水平最高。通过 Targetscan，KSR2 mRNA 被预测为 miR-3138 的靶标。并通过 qRT-PCR 和 Western blot 分别检测 miR-3138、KSR2、GLUT4、AMPK、PI3K、Akt 的 mRNA 和蛋白表达水平，通过双荧光素酶报告基因实验评估 miR-3138 和 KSR2 之间的相互作用。我们的结果表明，miR-3138 通过 KSR2/AMPK/GLUT4 信号通路显著加重了 HUVEC 的 IR。
