Department of Endocrinology, The Second Affiliated Hospital of Shandong First Medical University, Tai'an, China.
Department of General Medicine, The Second Affiliated Hospital of Shandong First Medical University, Tai'an, China.
Immun Inflamm Dis. 2024 Mar;12(3):e1191. doi: 10.1002/iid3.1191.
Diabetic cardiomyopathy (DCM) represents a major cause of heart failure and a large medical burden worldwide. This study screened the potentially regulatory targets of DCM and analyzed their roles in high glucose (HG)-induced cardiomyocyte injury.
Through GEO database, we obtained rat DCM expression chips and screened differentially expressed genes. Rat cardiomyocytes (H9C2) were induced with HG. 3-hydroxy-3-methylglutarylcoenzyme A synthase 2 (Hmgcs2) and microRNA (miR)-363-5p expression patterns in cells were measured by real-time quantitative polymerase chain reaction or Western blot assay, with the dual-luciferase assay to analyze their binding relationship. Then, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay, lactate dehydrogenase assay, terminal deoxynucleotidyl transferase dUTP nick end labeling assay, enzyme-linked immunosorbent assay, and various assay kits were applied to evaluate cell viability, cytotoxicity, apoptosis, inflammation responses, and oxidative burden.
Hmgcs2 was the vital hub gene in DCM. Hmgcs2 was upregulated in HG-induced cardiomyocytes. Hmgcs2 downregulation increased cell viability, decreased TUNEL-positive cell number, reduced HG-induced inflammation and oxidative stress. miR-363-5p is the upstream miRNA of Hmgcs2. miR-363-5p overexpression attenuated HG-induced cell injury.
Hmgcs2 had the most critical regulatory role in DCM. We for the first time reported that miR-363-5p inhibited Hmgcs2 expression, thereby alleviating HG-induced cardiomyocyte injury.
糖尿病心肌病（DCM）是心力衰竭的主要原因，也是全球范围内的一个重大医疗负担。本研究筛选了 DCM 的潜在调控靶点，并分析了它们在高糖（HG）诱导的心肌细胞损伤中的作用。
通过 GEO 数据库，我们获得了大鼠 DCM 表达芯片，并筛选出差异表达基因。用 HG 诱导大鼠心肌细胞（H9C2）。实时定量聚合酶链反应或 Western blot 检测细胞中 3-羟基-3-甲基戊二酰基辅酶 A 合酶 2（Hmgcs2）和 microRNA（miR）-363-5p 的表达模式，用双荧光素酶报告基因分析它们的结合关系。然后，应用 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐法、乳酸脱氢酶法、末端脱氧核苷酸转移酶 dUTP 缺口末端标记法、酶联免疫吸附测定法和各种试剂盒评估细胞活力、细胞毒性、细胞凋亡、炎症反应和氧化应激。
Hmgcs2 是 DCM 的关键枢纽基因。HG 诱导的心肌细胞中 Hmgcs2 上调。Hmgcs2 下调可增加细胞活力，减少 TUNEL 阳性细胞数量，减轻 HG 诱导的炎症和氧化应激。miR-363-5p 是 Hmgcs2 的上游 miRNA。miR-363-5p 过表达可减轻 HG 诱导的细胞损伤。
Hmgcs2 在 DCM 中具有最重要的调控作用。我们首次报道 miR-363-5p 抑制 Hmgcs2 表达，从而减轻 HG 诱导的心肌细胞损伤。
