Department for Pathophysiology, Centre for Research on Nutrition, Metabolism and Diabetes, Third Faculty of Medicine, Charles University, Prague, Czechia.
Franco-Czech Laboratory for Clinical Research on Obesity, Third Faculty of Medicine, Prague, and Inserm, Toulouse, France.
Front Endocrinol (Lausanne). 2021 Feb 15;11:582732. doi: 10.3389/fendo.2020.582732. eCollection 2020.
Development of type 2 diabetes (T2DM) is associated with disturbances in immune and metabolic status that may be reflected by an altered gene expression profile of peripheral blood mononuclear cells (PBMC). To reveal a potential family predisposition to these alterations, we investigated the regulation of gene expression profiles in circulating CD14 and CD14 PBMC in fasting conditions and in response to oral glucose tolerance test (OGTT) in glucose tolerant first-degree relatives (FDR) of T2DM patients and in control subjects.
This work is based on the clinical study LIMEX (NCT03155412). Non-obese 12 non-diabetic (FDR), and 12 control men without family history of diabetes matched for age and BMI underwent OGTT. Blood samples taken before and at the end of OGTT were used for isolation of circulating CD14 and CD14 PBMC. In these cells, mRNA levels of 94 genes related to lipid and carbohydrate metabolism, immunity, and inflammation were assessed by qPCR.
Irrespectively of the group, the majority of analyzed genes had different mRNA expression in CD14 PBMC compared to CD14 PBMC in the basal (fasting) condition. Seven genes (IRS1, TLR2, TNFα in CD14 PBMC; ABCA1, ACOX1, ATGL, IL6 in CD14 PBMC) had different expression in control vs. FDR groups. OGTT regulated mRNA levels of nine genes selectively in CD14 PBMC and of two genes (ABCA1, PFKL) selectively in CD14PBMC. Differences in OGTT-induced response between FDR and controls were observed for EGR2, CCL2 in CD14 PBMC and for ABCA1, ACOX1, DGAT2, MLCYD, and PTGS2 in CD14 PBMC.
This study revealed a different impact of glucose challenge on gene expression in CD14 when compared with CD14 PBMC fractions and suggested possible impact of family predisposition to T2DM on basal and OGTT-induced gene expression in these PBMC fractions. Future studies on these putative alterations of inflammation and lipid metabolism in fractionated PBMC in larger groups of subjects are warranted.
2 型糖尿病（T2DM）的发生与免疫和代谢状态的紊乱有关，这些变化可能反映在外周血单个核细胞（PBMC）的基因表达谱改变上。为了揭示这些变化的潜在家族易感性，我们研究了空腹状态下和口服葡萄糖耐量试验（OGTT）后 T2DM 患者一级亲属（FDR）和对照者循环 CD14 和 CD14 PBMC 中基因表达谱的调节。
本工作基于临床研究 LIMEX（NCT03155412）。12 名非肥胖、非糖尿病的非糖尿病 FDR（n=12）和 12 名年龄和 BMI 匹配、无糖尿病家族史的对照者接受 OGTT。OGTT 前后采集的血样用于分离循环 CD14 和 CD14 PBMC。在这些细胞中，通过 qPCR 评估与脂质和碳水化合物代谢、免疫和炎症相关的 94 个基因的 mRNA 水平。
无论组别如何，与基础（空腹）状态下的 CD14 PBMC 相比，大多数分析的基因在 CD14 PBMC 中的 mRNA 表达不同。在 CD14 PBMC 中，有 7 个基因（IRS1、TLR2、TNFα）和 CD14 PBMC 中的 2 个基因（ABCA1、ACOX1）在对照组与 FDR 组之间的表达不同。OGTT 选择性调节了 CD14 PBMC 中 9 个基因和 CD14 PBMC 中 2 个基因（ABCA1、PFKL）的 mRNA 水平。FDR 与对照组之间在 CD14 PBMC 中 EGR2、CCL2 和 CD14 PBMC 中 ABCA1、ACOX1、DGAT2、MLCYD、PTGS2 的 OGTT 诱导反应存在差异。
本研究揭示了葡萄糖刺激对 CD14 与 CD14 PBMC 分数中基因表达的不同影响，并提示 T2DM 的家族易感性可能对这些 PBMC 分数中的基础和 OGTT 诱导的基因表达有影响。需要在更大的受试者群体中对这些有争议的炎症和脂代谢变化进行进一步研究。
