Zhang LingHui, Zhang ShengLi
Department of Endocrinology, Hubei Third People's Hospital affiliated to Jianghan University, Wuhan City, China.
Department of Cardiovascular Medicine, Hubei Third People's Hospital affiliated to Jianghan University, Wuhan City, China.
Turk J Biol. 2022 Apr 25;46(4):298-306. doi: 10.55730/1300-0152.2618. eCollection 2022.
Diabetes resulting from insufficient insulin secretion or insulin resistance (IR) is a highly prevalent metabolic disease. Since microRNAs have been linked with elevated IR, the current research hypothesized that miR-124-3p has a role in IR and the establishment of IR and type 2 diabetes (T2DM). The study aimed to explore the molecular mechanisms of miR-124-3p which influence IR leading to T2DM establishment. HepG2 cells were cultured in vitro, and palmitic acid (PA) was used to construct the IR cell model. In the IR model, transfection of miR-124-3p or phosphoglucomutase 1 (PGM1) linked plasmids were transfected into HepG2 cells. RT-qPCR was used to determine the miR-124-3p and PGM1 expressions in the cells. Cell viability was assessed through CCK-8 assays, while glucose consumption was studied using a glucose uptake test. Interaction between miR-124-3p and PGM1 was examined using a dual-luciferase reporter assay. Autophagy, phosphatidylinositol 3 kinases (PI3K)/protein kinase B (AKT) and JNK pathways-linked factors, glucose transporter 4 (GLUT4), and c-Jun were determined through western blotting assays. MiR-124-3p expression was elevated, but PGM1 was reduced in the IR model. Glucose uptake was reduced posttreatment with 0.8 mM PA. There was a significantly increased PI3K, p-PI3K, AKT, p-AKT, GLUT4, LC3I/II, Beclin-1, p-JNK1/2, and c-Jun, but reduced p62 expressions were presented in the PA + miR-124-3p inhibitor compared to the PA and PA + inhibitor NC groups. PGM1 binds directly to miR-124-3p through the 3' UTR region target. Overall, miR-124-3p downregulates glucose consumption via targeting PGM1 to repress PI3K/AKT and JNK pathways. Silencing PGM1 inhibited the suppressor role of miR-124-3p on glucose uptake, cell proliferation, and inflammation. In conclusion, miR-124-3p reduces glucose uptake in HepG2 cells via PGM1/PI3K/AKT modulation. MiR-124-3p targets PGM1 in IR and may provide an effective therapeutic alternative for T2DM.
胰岛素分泌不足或胰岛素抵抗（IR）导致的糖尿病是一种高度流行的代谢性疾病。由于微小RNA与IR升高有关，当前研究假设miR-124-3p在IR以及IR和2型糖尿病（T2DM）的发生中起作用。该研究旨在探索miR-124-3p影响IR导致T2DM发生的分子机制。体外培养HepG2细胞，并用棕榈酸（PA）构建IR细胞模型。在IR模型中，将miR-124-3p或磷酸葡萄糖变位酶1（PGM1）相关质粒转染到HepG2细胞中。采用RT-qPCR测定细胞中miR-124-3p和PGM1的表达。通过CCK-8法评估细胞活力，同时使用葡萄糖摄取试验研究葡萄糖消耗情况。采用双荧光素酶报告基因检测法检测miR-124-3p与PGM1之间的相互作用。通过蛋白质印迹法检测自噬、磷脂酰肌醇3激酶（PI3K）/蛋白激酶B（AKT）和JNK通路相关因子、葡萄糖转运蛋白4（GLUT4）和c-Jun。在IR模型中，miR-124-3p表达升高，但PGM1表达降低。用0.8 mM PA处理后葡萄糖摄取减少。与PA组和PA +抑制剂NC组相比，PA + miR-124-3p抑制剂组中PI3K、p-PI3K、AKT、p-AKT、GLUT4、LC3I/II、Beclin-1、p-JNK1/2和c-Jun显著增加，但p62表达降低。PGM1通过3'UTR区域靶点直接与miR-124-3p结合。总体而言，miR-124-3p通过靶向PGM1下调葡萄糖消耗，从而抑制PI3K/AKT和JNK通路。沉默PGM1可抑制miR-124-3p对葡萄糖摄取、细胞增殖和炎症的抑制作用。总之，miR-124-3p通过PGM1/PI3K/AKT调节降低HepG2细胞中的葡萄糖摄取。miR-124-3p在IR中靶向PGM1，可能为T2DM提供一种有效的治疗选择。
