Department of Biology, Faculty of Philosophy, Sciences and Letters of Ribeirão Preto, University of São Paulo - USP, Ribeirão Preto, SP, Brazil.
Gene. 2012 Dec 15;511(2):151-60. doi: 10.1016/j.gene.2012.09.090. Epub 2012 Oct 2.
Patients with type 2 diabetes mellitus (T2DM) exhibit insulin resistance associated with obesity and inflammatory response, besides an increased level of oxidative DNA damage as a consequence of the hyperglycemic condition and the generation of reactive oxygen species (ROS). In order to provide information on the mechanisms involved in the pathophysiology of T2DM, we analyzed the transcriptional expression patterns exhibited by peripheral blood mononuclear cells (PBMCs) from patients with T2DM compared to non-diabetic subjects, by investigating several biological processes: inflammatory and immune responses, responses to oxidative stress and hypoxia, fatty acid processing, and DNA repair. PBMCs were obtained from 20 T2DM patients and eight non-diabetic subjects. Total RNA was hybridized to Agilent whole human genome 4×44K one-color oligo-microarray. Microarray data were analyzed using the GeneSpring GX 11.0 software (Agilent). We used BRB-ArrayTools software (gene set analysis - GSA) to investigate significant gene sets and the Genomica tool to study a possible influence of clinical features on gene expression profiles. We showed that PBMCs from T2DM patients presented significant changes in gene expression, exhibiting 1320 differentially expressed genes compared to the control group. A great number of genes were involved in biological processes implicated in the pathogenesis of T2DM. Among the genes with high fold-change values, the up-regulated ones were associated with fatty acid metabolism and protection against lipid-induced oxidative stress, while the down-regulated ones were implicated in the suppression of pro-inflammatory cytokines production and DNA repair. Moreover, we identified two significant signaling pathways: adipocytokine, related to insulin resistance; and ceramide, related to oxidative stress and induction of apoptosis. In addition, expression profiles were not influenced by patient features, such as age, gender, obesity, pre/post-menopause age, neuropathy, glycemia, and HbA(1c) percentage. Hence, by studying expression profiles of PBMCs, we provided quantitative and qualitative differences and similarities between T2DM patients and non-diabetic individuals, contributing with new perspectives for a better understanding of the disease.
2 型糖尿病（T2DM）患者除了表现出与肥胖和炎症反应相关的胰岛素抵抗外，还表现出高水平的氧化 DNA 损伤，这是高血糖和活性氧（ROS）产生的结果。为了提供有关 T2DM 病理生理学中涉及的机制的信息，我们分析了 T2DM 患者与非糖尿病患者外周血单核细胞（PBMC）的转录表达模式，研究了几个生物学过程：炎症和免疫反应、氧化应激和缺氧反应、脂肪酸处理和 DNA 修复。从 20 名 T2DM 患者和 8 名非糖尿病患者中获得 PBMC。将总 RNA 杂交到 Agilent 全人类基因组 4×44K 单色寡聚微阵列上。使用 GeneSpring GX 11.0 软件（Agilent）分析微阵列数据。我们使用 BRB-ArrayTools 软件（基因集分析-GSA）来研究显著的基因集，并用 Genomica 工具来研究临床特征对基因表达谱的可能影响。我们表明，与对照组相比，T2DM 患者的 PBMC 表现出明显的基因表达变化，有 1320 个差异表达基因。大量基因参与了 T2DM 发病机制中涉及的生物学过程。在具有高倍数变化值的基因中，上调的基因与脂肪酸代谢和脂质诱导的氧化应激保护有关，而下调的基因与抑制促炎细胞因子产生和 DNA 修复有关。此外，我们确定了两个重要的信号通路：与胰岛素抵抗相关的脂肪细胞因子；与氧化应激和细胞凋亡诱导相关的神经酰胺。此外，表达谱不受患者特征（如年龄、性别、肥胖、绝经前后年龄、神经病变、血糖和 HbA1c%）的影响。因此，通过研究 PBMC 的表达谱，我们提供了 T2DM 患者与非糖尿病个体之间的定量和定性差异和相似之处，为更好地理解该疾病提供了新的视角。
