Circ_MEF2D /miR-361/AMPK作用轴在血管内皮前体细胞治疗糖尿病足中的作用机制研究

Diabetic foot which Lacking of effective treatment methods is the main cause of disability in diabetes mellitus. Injurying of endothelial progenitor cells (EPCs) is an independent risk factor for vascular complications in diabetes mellitus. Previous studies in our team have confirmed that EPCs can differentiate into mature vascular endothelial cells in vitro and have therapeutic effects in diabetic foot model after transplantation of EPCs into the rat foot. However the mechanism is not clear. In our preliminary experiment, we analyzed the tissue of animal model by gene sequencing and RT-qPCR. We found that the expression of circ_MEF2D in diabetic foot decreased significantly, and the expression of circ_MEF2D increased significantly after EPCs treatment. The target gene of mir-361 was predicted by the database. Further cluster enrichment analysis revealed that the adenosine monophosphate-activated protein kinase (AMPK) pathway was related to the EPCs treatment. Thus, we postulate that EPCs may treat diabetic foot through circ_MEF2D/mir-361/AMPK axis. This study aims to elucidate the above hypothesis on gene, protein, cell and animal levels, using gene knockdown and overexpression, RNA pull-down, RNA immunoprecipitation and other methods, so as to provide new theoretical basis for preventing and treating diabetic angiopathy.

糖尿病足（DF）是糖尿病的主要致残原因，缺少有效治疗方法，血管内皮前体细胞（EPCs）损伤是糖尿病血管并发症的独立危险因素。本课组先前研究证实移植EPCs到DF大鼠模型足部有治疗作用，但其机制尚不明确。我们预实验通过基因测序、RT-qPCR对动物模型组织进行分析，发现DF中circ_MEF2D的表达量显著降低，EPCs治疗后circ_MEF2D的表达量显著升高，通过数据库预测miR-361为其靶基因，进一步通过聚类富集分析发现腺苷酸活化的蛋白激酶（AMPK）通路与EPCs治疗有关。据此我们提出假说：EPCs可能通过 circ_MEF2D/miR-361/AMPK作用轴治疗DF。本研究拟从基因、蛋白、细胞和动物水平，利用基因敲减和过表达、RNA pull-down、RNA免疫共沉淀等方法，阐明上述假说，从而为糖尿病血管病变的防治提供新思路。

糖尿病足（DF）是糖尿病的主要致残原因，缺少有效治疗方法，血管内皮前体细胞（EPCs）损伤是糖尿病血管并发症的独立危险因素。本课题组先前研究证实移植EPCs到DF大鼠模型足部有治疗作用，但其机制尚不明确。预实验通过基因测序、RT-qPCR对动物模型组织进行分析，发现DF中circ_MEF2D的表达量显著降低，EPCs治疗circ_MEF2D的表达量显著升高。通过数据库预测miR-361为其靶基因，进一步通过聚类富集分析发现腺苷酸活化的蛋白激酶（AMPK）通路与EPCs治疗有关。本研究从基因、蛋白、细胞和动物水平，利用基因敲减和过表达、RNA pull-down、RNA免疫共沉淀等方法证明EPCs通过 circ_MEF2D/miR-361/AMPK作用轴治疗DF，为糖尿病血管病变的防治提供科学依据。并继续进行了DM和DF患者circRNA和mRNA谱及其相互作用的联合分析，发现circRNA CNOT6、AXIN1以及mRNA STAT3、MYD88和B2M，Nod样受体信号通路、胰岛素信号通路、鞘磷脂代谢通路和核糖体通路；circRNA FBXO7、ATM和LMBRD1，mRNA RUNX1和TLR6，溶酶体通路、Chagas病通路和甲烷代谢通路在DM和DF的发生发展中同样起重要作用，为DF的发生发展机制提供了新的分子理论基础。为了探索糖代谢异常引起相关代谢紊乱的机制，进行了肝脏、肌肉脂肪变性的氨基酸传感器的鉴定。发现作为一种脂肪降解的抑制剂，Plin2在脂肪变性患者的肝脏和肌肉中显著上调，Plin2中的任何赖氨酸残基都可以被泛素连接，并进一步证明了Plin2的泛素化由E3泛素连接酶Ubr1完成，Ubr1通过感受氨基酸变化来调控Plin2的稳定性。这些发现填补了营养感知和脂肪变性之间联系的空白，可能为代谢相关疾病的营养治疗提供新的策略。

内容获取失败，请点击重试