通过转录组测序分析识别糖尿病患者冠状动脉粥样硬化的新途径

Transcriptome sequencing analysis in patients with diabetes to identify coronary heart disease

摘要:

目的 探究冠状动脉疾病（CAD）和2型糖尿病（T2DM）患者心外膜脂肪组织（EAT）的转录组变化。方法 选取青海省心脑血管病专科医院8例CAD患者，并根据有无T2DM将患者分为糖尿病组（D组，n?=5）和非糖尿病组（ND组，n?=3）。对8例患者EAT样本进行RNA测序分析。基因富集分析以鉴定受差异表达基因影响的途径。通过逆转录聚合酶链反应（RT-PCR）及Western blotting检测验证靶基因的变化。结果 有592个基因在D组EAT中差异表达。在D组EAT中差异表达基因主要富集于炎症相关反应，例如CSF3、IL-1β，IL-6。KEGG通路分析证实，上调的基因参与炎症途径，如肿瘤坏死因子（TNF），NF-κB及AGE-RAGE。在氧化应激途径中通过实验验证两个差异有统计学意义的基因（FRA1和PTX3）。结论 糖尿病EAT的转录组变化与高血糖引起的氧化应激密切相关，其基因的差异表达可能激活了氧化应激反应并促进糖尿病患者的动脉粥样硬化。

Abstract:

Objective To evaluate the epicardial adipose tissue (EAT) transcriptome analysis in coronary artery disease (CAD) complicated with type 2 diabetes (T2DM). Methods EAT samples were obtained from subjects with CAD patients and they were divided into diabetes group (D group, n?=?5) and non-T2DM (ND group, n?=?3). RNA-sequencing analysis was performed in EAT. Gene enrichment analysis was conducted to identify differentially expressed signal pathways which were verified by quantitative RT-PCR (qRT-PCR) and western blotting. Results A total of 592 genes were differentially expressed in diabetic EAT, which were mainly inflammatory genes including Colony Stimulating Factor 3 (CSF3), Interleukin-1β (IL-1β) and IL-6. KEGG pathway analysis confirmed that upregulated genes were involved in inflammatory pathways including Tumor Necrosis Factor (TNF), Nuclear Factor-κB (NF-κB) and advanced glycation end-products-receptor advanced glycation end products (AGE-RAGE). FRA1 and PTX3 were experimentally validated in an oxidative stress pathway. Conclusions The transcriptome changes of diabetic EAT are closely related to the hyperglycemia-induced oxidative stress, which may activate oxidative stress and promote atherosclerosis in diabetic patients.