Department of Nephrology, The First Hospital of Jilin University, Changchun 130000, Jilin, P.R. China.
Department of Andrology, The First Hospital of Jilin University, Changchun 130000, Jilin, P.R. China.
Biosci Rep. 2019 Jul 2;39(7). doi: 10.1042/BSR20181852. Print 2019 Jul 31.
The present study aims to investigate the mechanism of miR-15a-5p in the atherosclerotic (AS) inflammatory response and arterial injury improvement in diabetic rats by regulating fatty acid synthase (FASN). Initially, bioinformatics tools were applied to evaluate miRNAs and genes correlating with AS, and the target relation between miRNAs and FASN was measured using the Dual-Luciferase Reporter Assay. Subsequently the diabetic AS rat model was established and the surviving rats were divided into: negative control (NC), miR-15a-5p mimic, miR-15a-5p inhibitor, sh-FASN and miR-15a-5p + sh-FASN groups. Then a series of experiments were performed to examine the degree of AS in each group. The results revealed that compared with the NC group, the expressions of C-reactive protein (CRP), interleukin 6 (IL-6), intercellular cell adhesion molecule-1 (ICAM1) in rat arterial tissue, as well as the levels of low-density lipoprotein cholesterol (LDL-C), blood glucose (BG), triglycerides (TG), total cholesterol (TC) and Homocysteine (Hcy) in rat serum, were increased after inhibiting miR-15a-5p, while the level of high-density lipoprotein cholesterol (HDL-C) was decreased and the fat storage area was enlarged after this treatment (<0.05). In the miR-15a-5p mimic and sh-FASN groups, serum HDL-C levels were increased and the fat storage areas in arteries were reduced. The levels of CRP, IL-6, ICAM1 in rat arterial tissue, along with the levels of LDL-C, BG, TG, TC and Hcy in rat serum, were decreased (<0.05). Hematoxylin and Eosin (HE) staining and transmission electron microscopy (TEM) results showed AS lesions to be apparent in the arteries of rats in both the NC and miR-15a-5p inhibitor groups, but that in miR-15a-5p and sh-FASN group were improved, the miR-15a-5p mimic + sh-FASN group showed the most obvious improvement. Taken together, miR-15a-5p alleviates the inflammation response and arterial injury in diabetic AS rats by targeting FASN.
本研究旨在通过调节脂肪酸合酶（FASN）来探讨 miR-15a-5p 在糖尿病大鼠动脉粥样硬化（AS）炎症反应和动脉损伤改善中的作用机制。首先，应用生物信息学工具评估与 AS 相关的 miRNA 和基因，并使用双荧光素酶报告基因实验测量 miRNA 与 FASN 的靶关系。随后，建立糖尿病 AS 大鼠模型，将存活大鼠分为阴性对照组（NC）、miR-15a-5p 模拟物组、miR-15a-5p 抑制剂组、sh-FASN 组和 miR-15a-5p+sh-FASN 组。然后进行一系列实验来检测每组大鼠的 AS 程度。结果表明，与 NC 组相比，抑制 miR-15a-5p 后，大鼠动脉组织中 C 反应蛋白（CRP）、白细胞介素 6（IL-6）、细胞间黏附分子 1（ICAM1）的表达，以及大鼠血清中低密度脂蛋白胆固醇（LDL-C）、血糖（BG）、甘油三酯（TG）、总胆固醇（TC）和同型半胱氨酸（Hcy）的水平均升高，而高密度脂蛋白胆固醇（HDL-C）水平降低，脂肪储存面积增大（<0.05）。在 miR-15a-5p 模拟物和 sh-FASN 组中，血清 HDL-C 水平升高，动脉脂肪储存面积减少。大鼠动脉组织中 CRP、IL-6、ICAM1 的表达，以及大鼠血清中 LDL-C、BG、TG、TC 和 Hcy 的水平均降低（<0.05）。苏木精-伊红（HE）染色和透射电镜（TEM）结果显示，NC 组和 miR-15a-5p 抑制剂组大鼠动脉均出现明显的 AS 病变，而 miR-15a-5p 和 sh-FASN 组病变得到改善，miR-15a-5p 模拟物+sh-FASN 组改善最明显。综上所述，miR-15a-5p 通过靶向 FASN 减轻糖尿病 AS 大鼠的炎症反应和动脉损伤。
