School of the Environment and Safety Engineering, Jiangsu University, Zhenjiang, Jiangsu 212013, PR China; Institute of Life Sciences, Jiangsu University, Zhenjiang, Jiangsu 212013, PR China.
The Fourth Affiliated Hospital of Jiangsu University, 20# Zhengdong Road, Zhenjiang, Jiangsu 212001, PR China.
J Diabetes Complications. 2020 Feb;34(2):107444. doi: 10.1016/j.jdiacomp.2019.107444. Epub 2019 Sep 13.
Diabetes mellitus (DM) is a chronic disease characterized by hyperglycemia resulting from defects in insulin secretion, insulin action, or both. The liver is a key organ involved in glucose metabolism, and the major target proteins' changes in the pathogenesis are still unknown.
A diabetic mouse model was induced by intraperitoneal injection of streptozotocin (STZ) solution and the RNA-Seq analysis was used to evaluate the transcription differences in the livers of diabetic mice of this study. And then, the differentially expressed genes were validated between a normal mouse group (n = 6) and a diabetic mouse group (n = 6) using quantitative real-time PCR (qRT-PCR) and Western blotting analysis. In addition, we also constructed protein-protein interacting (PPI) networks of up-regulated and down-regulated genes.
Transcriptome sequencing analysis revealed 370 up-regulated differentially expressed genes and 281 down-regulated differentially expressed genes in the diabetes model. The gene ontology (GO) analysis and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis results showed that the differentially expressed genes were mainly involved in immunity, enzyme activity, metabolism, and steroid synthesis. PPI analysis results indicated that the main 15 core differential proteins (Cyp51a1, Acsl4, Ugt1a1, Stat1, Gsta2, Cbr1, Aldh1a1, Fasn, Ces1, Camk2b, Tap1, Egr1, Sqle, Lpin1, Fabp5) were involved in the pathogenesis of diabetes. The qRT-PCR results showed that expression changes of four genes (Acsl4, Stat1, Gsta2, Fabp5) were in different directions from those of RNA-Seq. Western blotting results indicated that Sqle expression change at the protein level was in opposition direction from qRT-PCR, and we speculated that Sqle may be involved in the post-transcriptional modification process.
Our data speculated that the pathogenesis of diabetes may be mediated mainly through steroid biosynthesis, metabolic processes, and immune responses. Further researches on these pathways may provide new targets for the prevention and treatment of diabetes.
糖尿病（DM）是一种以胰岛素分泌缺陷或胰岛素作用缺陷为特征的慢性疾病，导致血糖升高。肝脏是参与葡萄糖代谢的关键器官，其主要靶蛋白在发病机制中的变化仍不清楚。
通过腹腔注射链脲佐菌素（STZ）溶液诱导糖尿病小鼠模型，使用 RNA-Seq 分析评估本研究中糖尿病小鼠肝脏的转录差异。然后，使用定量实时 PCR（qRT-PCR）和 Western blot 分析验证正常小鼠组（n=6）和糖尿病小鼠组（n=6）之间差异表达基因的验证。此外，我们还构建了上调和下调基因的蛋白质-蛋白质相互作用（PPI）网络。
转录组测序分析显示，糖尿病模型中 370 个上调差异表达基因和 281 个下调差异表达基因。基因本体（GO）分析和京都基因与基因组百科全书（KEGG）通路富集分析结果表明，差异表达基因主要涉及免疫、酶活性、代谢和类固醇合成。PPI 分析结果表明，主要的 15 个核心差异蛋白（Cyp51a1、Acsl4、Ugt1a1、Stat1、Gsta2、Cbr1、Aldh1a1、Fasn、Ces1、Camk2b、Tap1、Egr1、Sqle、Lpin1、Fabp5）参与了糖尿病的发病机制。qRT-PCR 结果显示，四个基因（Acsl4、Stat1、Gsta2、Fabp5）的表达变化与 RNA-Seq 的结果不一致。Western blot 结果表明，Sqle 蛋白水平表达变化与 qRT-PCR 结果相反，我们推测 Sqle 可能参与了转录后修饰过程。
我们的数据推测，糖尿病的发病机制可能主要通过类固醇生物合成、代谢过程和免疫反应来介导。对这些通路的进一步研究可能为糖尿病的预防和治疗提供新的靶点。
