内皮祖细胞DNA羟甲基化修饰在糖尿病血管并发症发病中的作用及其机制-现代医学

内皮祖细胞DNA羟甲基化修饰在糖尿病血管并发症发病中的作用及其机制

单位：华中科技大学同济医学院附属武汉中心医院内分泌科, 湖北武汉 430014

分类号：R587.2

出版年·卷·期（页码）：2021·49·第一期（1-5）

摘要：

目的： 检测糖尿病患者外周血内皮祖细胞（endothelial progenitor cells，EPCs）黏附功能及其DNA的羟甲基化水平，并探讨DNA羟甲基化水平与糖尿病血管并发症患者EPCs功能的关系。方法： 采用梯度离心法分离30例健康志愿者（正常对照组）和30例2型糖尿病伴有血管并发症患者（糖尿病组）的外周血单个核细胞，并利用EBM-2培养基将其诱导分化为EPCs。检测细胞黏附纤维蛋白的能力判断EPCs的功能是否异常。采用DNA提取试剂盒提取EPCs DNA，通过液相色谱-串联质谱分析法测定5-羟甲基胞嘧啶（5hmC）和5-甲基胞嘧啶（5mC）含量，判断EPCs DNA的羟甲基化水平。结果：与正常对照组相比，糖尿病组EPCs的黏附能力显著下降（P<0.05）；EPCs DNA的羟甲基化修饰水平明显提高；5hmC和5mC含量显著下降（P<0.01及P<0.05），两者的比值明显降低（P<0.05）。结论： 糖尿病伴血管并发症患者的外周血EPCs功能显著下降，可能与DNA羟甲基化修饰有关，提示DNA羟甲基化修饰导致EPCs功能下降可能是糖尿病血管并发症发生的重要机制。

Objective: To detected adhesion function of endothelial progenitor cells(EPCs) and the level of hydroxylation of DNA in the peripheral blood group of diabetic peripheral blood, and to discuss the relationship between the level of DNA hydroxylation and the EPCs function of diabetic vascular complications. Methods: The peripheral blood mononuclear cells were isolated from 30 healthy volunteers and 30 patients with diabetes mellitus with angiopathy by gradient centrifugation, and the EBM-2 medium was used to induce the cells to differentiate into EPCs. The function of EPCs was evaluated by detecting the ability of cell adhesion to fibrin. EPCs DNA was extracted by DNA extraction kit, and the contents of 5-hydroxymethylcytosine(5hmC) and 5-methylcytosine (5mC) were determined by liquid chromatography tandem mass spectrometry, and the level of hydroxyl methylation of EPCs DNA was determined. Results: The adhesion ability of EPCs in diabetic group decreased significantly(P<0.05) compared with that in the control group, and the content of 5 hmC and 5 mC decreased significantly(P<0.01, P<0.05). Conclusion: The EPCs function in peripheral blood of diabetic patients with vascular complications is significantly decreased, which may be associated with DNA methylation modification, suggesting that the decrease of EPCs function caused by DNA hydroxymethylation might be an important mechanism of diabetic vascular complications.

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