循环miRNA作为糖尿病脑病诊断标志物的研究

Circulating miRNA as a diagnostic marker of diabetic encephalopathy

摘要:

目的：研究糖尿病脑病外周血血清循环miRNA的表达情况,探讨miRNA-368对糖尿病脑病的预测效果。方法： qRT-PCR检测miRNA-368在糖尿病脑病中外周血血清的表达,并将miRNA-368转染到糖尿病脑病人外周血单核细胞中,然后通过qRT-PCR检测转染效率。采取CCK-8和克隆形成实验检测miRNA-368对糖尿病脑病人外周血单核细胞增殖的影响,流式细胞技术检测miRNA-368对糖尿病脑病人外周血单核细胞周期和凋亡率的影响。结果：27例患者糖尿病脑病组织miRNA-368的相对表达量为1.402,95% CI为0.363～0.595。miRNA-368相对表达量与患者的一般资料如年龄、性别并不存在相关性(P＞0.05)；而糖尿病血糖脑病患者的miRNA-368相对表达量明显上调(P＜0.05),而无糖尿病患者的相对表达量与正常组相比差异无统计学意义(P＞0.05),miRNA-368相对表达量明显上调与糖尿病脑病之间具有有关；qRT-PCR显示,糖尿病脑病组织和糖尿病脑病人外周血单核细胞miRNA-368的表达明显上调。si-miRNA-368干预后,糖尿病脑病人外周血单核细胞增殖受到抑制,凋亡率增加。结论：miRNA-368在糖尿病脑病人外周血单核细胞中的表达明显上调,促进了糖尿病脑病的发生发展。

Abstract:

Objective: To study the expression of miRNAs in peripheral blood of diabetic encephalopathy and to investigate the predicting effect of miRNA-368 on diabetic encephalopathy. Methods: qRT-PCR was used to detect the expression of miRNA-368 in peripheral blood of patients with diabetic encephalopathy in Sanya. MiRNA-368 was transfected into peripheral blood mononuclear cells of diabetic patients. The transfection efficiency was detected by qRT-PCR. The effect of miRNA-368 on the proliferation of peripheral blood mononuclear cells in diabetic brain patients was detected by CCK-8 and clonogenic assay. The effect of miRNA-368 on the peripheral blood mononuclear cell cycle and apoptotic rate in diabetic patients with cerebral encephalopathy was analyzed by flow cytometry. Results: The relative expression of miRNA-368 in 27 patients with diabetic encephalopathy was 1.402, and the 95% CI was 0.363-0.595. The relative expression level of miRNA-368 was not related to the general data of patients such as age and gender (P> 0.05), while the relative expression of miRNA-368 in patients with diabetic encephalopathy was significantly increased (P <0.05). The relative expression of miRNA-368 was significantly correlated with that of diabetic encephalopathy. QRT-PCR showed that peripheral blood of patients with diabetic encephalopathy and diabetic encephalopathy Monocyte miRNA-368 expression was significantly up-regulated. After the intervention of si-miRNA-368, the proliferation of peripheral blood mononuclear cells in diabetic brain patients was inhibited and the apoptosis rate increased. Conclusion: The expression of miRNA-368 in peripheral blood mononuclear cells of diabetic encephalopathy patients was significantly up-regulated, which promoted the development of diabetic encephalopathy.