Jin Guoxi, Wang Qiong, Pei Xiaoyan, Li Xiaoli, Hu Xiaolei, Xu Erqin, Li Minglong
Department of Endocrinology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, Shandong 250021, People's Republic of China.
Department of Endocrinology, The First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui 233004, People's Republic of China.
Diabetes Metab Syndr Obes. 2019 Jul 23;12:1249-1261. doi: 10.2147/DMSO.S206270. eCollection 2019.
A long-term "memory" of hyperglycemic stress, even when glycemia is normalized, has been previously reported in endothelial cells. However, the molecular mechanism of "metabolic memory" (MM) remains unknown. In this report, we sought to screen at the whole transcriptome level the genes that participate in MM.
In the present research, RNA sequencing was used to determine the protein-coding mRNA expression profiles of human umbilical vein endothelial cells (HUVECs) under normal-glucose concentration (LG), high-glucose concentration (HG), and MM. A series of bioinformatic analyses was performed. HG-induced MM-involved up-regulated genes (up-HGMMGs) and HG-induced MM-involved down-regulated genes (down-HGMMGs) were identified. Afterward, based on up-HGMMGs and down-HGMMGs, the biological functions and signaling pathways were analyzed using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG). In addition, several of the identified genes were validated by RT-qPCR.
A total of 726 HGMMGs were identified, including 210 down- and 516 up-HGMMGs, which were enriched in the cell cycle (hsa04110), oocyte meiosis (hsa04114), p53 signaling pathway (hsa04115), and oxidative phosphorylation (hsa00190), among others. The protein-protein-interaction (PPI) network consisted of 462 nodes and 2656 connections, and four main modules were identified by MCODE. The cell cycle (hsa04110), oocyte meiosis (hsa04114), p53 signaling pathway (hsa04115), and oxidative phosphorylation (hsa00190), among others, could be potential therapeutic targets of HG-induced MM in endothelial cells. The real-time PCR results validated the RNA-seq data.
This study identified crucial mRNAs related to MM-persistent injury in endothelial cells even after switching the cells from high- glucose to normal glucose levels. Further research focusing on these mRNA may unravel new ways to modify MM in diabetes.
先前已有报道称，即使血糖恢复正常，内皮细胞中仍存在高血糖应激的长期“记忆”。然而，“代谢记忆”（MM）的分子机制仍不清楚。在本报告中，我们试图在全转录组水平筛选参与MM的基因。
在本研究中，RNA测序用于确定人脐静脉内皮细胞（HUVECs）在正常葡萄糖浓度（LG）、高葡萄糖浓度（HG）和MM条件下的蛋白质编码mRNA表达谱。进行了一系列生物信息学分析。鉴定出HG诱导的MM相关上调基因（up-HGMMGs）和HG诱导的MM相关下调基因（down-HGMMGs）。随后，基于up-HGMMGs和down-HGMMGs，使用基因本体论（GO）和京都基因与基因组百科全书（KEGG）分析其生物学功能和信号通路。此外，通过RT-qPCR对鉴定出的几个基因进行了验证。
共鉴定出726个HGMMGs，包括210个下调的和516个上调的HGMMGs，它们富集于细胞周期（hsa04110）、卵母细胞减数分裂（hsa04114）、p53信号通路（hsa04115）和氧化磷酸化（hsa00190）等通路。蛋白质-蛋白质相互作用（PPI）网络由462个节点和2656个连接组成，通过MCODE鉴定出四个主要模块。细胞周期（hsa04110）、卵母细胞减数分裂（hsa04114）、p53信号通路（hsa04115）和氧化磷酸化（hsa00190）等可能是HG诱导的内皮细胞MM的潜在治疗靶点。实时PCR结果验证了RNA-seq数据。
本研究鉴定出了与内皮细胞中MM持续性损伤相关的关键mRNA，即使在细胞从高糖水平转换为正常糖水平后依然存在。针对这些mRNA的进一步研究可能会揭示改善糖尿病中MM的新方法。
