Postgraduate Training Basement of Jinzhou Medical University, Taihe Hospital, Hubei University of Medicine, Shiyan, 442000, China.
Department of Cardiology, Taihe Hospital, Hubei University of Medicine, Shiyan, 442000, Hubei, China.
J Physiol Biochem. 2024 Feb;80(1):27-39. doi: 10.1007/s13105-023-00974-0. Epub 2023 Oct 4.
Cardiovascular disease due to atherosclerosis is one of the leading causes of death worldwide; however, the underlying mechanism has yet to be defined. The sodium-dependent glucose transporter 2 inhibitor (SGLT2i) empagliflozin is a new type of hypoglycemic drug. Recent studies have shown that empagliflozin not only reduces high glucose levels but also exerts cardiovascular-protective effects and slows the process of atherosclerosis. The purpose of this study was to elucidate the mechanism by which empagliflozin ameliorates atherosclerosis. Male apolipoprotein E-deficient (ApoE) mice were fed a high-fat Western diet to establish an atherosclerosis model. The area and size of atherosclerotic lesions in ApoE mice were then assessed by performing hematoxylin-eosin (HE) staining after empagliflozin treatment. Concurrently, oxidized low-density lipoprotein (oxLDL) was used to mimic atherosclerosis in three different types of cells. Then, following empagliflozin treatment of macrophage cells (RAW264.7), human aortic smooth muscle cells (HASMCs), and human umbilical vein endothelial cells (HUVECs), western blotting was applied to measure the levels of autophagy-related proteins and proinflammatory cytokines, and green fluorescent protein (GFP)-light chain 3 (LC3) puncta were detected using confocal microscopy to confirm autophagosome formation. Oil Red O staining was performed to detect the foaming of macrophages and HASMCs, and flow cytometry was used for the cell cycle analysis. 5-ethynyl-2'-deoxyuridine (EdU), cell counting kit-8 (CCK-8), and scratch assays were also performed to examine the proliferation and migration of HASMCs. Empagliflozin suppressed the progression of atherosclerotic lesions in ApoE mice. Empagliflozin also induced autophagy in RAW246.7 cells, HASMCs, and HUVECs via the adenosine monophosphate-activated protein kinase (AMPK) signaling pathway, and it significantly increased the levels of the Beclin1 protein, the LC3B-II/I ratio, and p-AMPK protein. In addition, empagliflozin decreased the expression of P62 and the protein levels of inflammatory cytokines, and it inhibited the foaming of RAW246.7 cells and HASMCs, as well as the expression of inflammatory factors by inducing autophagy. Empagliflozin activated autophagy through the AMPK signaling pathway to delay the progression of atherosclerosis. Furthermore, the results of flow cytometry, EdU assays, CCK-8 cell viability assays, and scratch assays indicated that empagliflozin blocked HASMCs proliferation and migration. Empagliflozin activates autophagy through the AMPK signaling pathway to delay the evolution of atherosclerosis, indicating that it may represent a new and effective drug for the clinical treatment of atherosclerosis.
由于动脉粥样硬化导致的心血管疾病是全球范围内主要的死亡原因之一；然而，其潜在的机制尚未确定。钠依赖性葡萄糖转运蛋白 2 抑制剂（SGLT2i）恩格列净是一种新型的降血糖药物。最近的研究表明，恩格列净不仅能降低高血糖水平，而且具有心血管保护作用，并能减缓动脉粥样硬化的进程。本研究旨在阐明恩格列净改善动脉粥样硬化的机制。雄性载脂蛋白 E 缺陷（ApoE）小鼠用高脂肪西方饮食喂养以建立动脉粥样硬化模型。然后通过对恩格列净处理后的载脂蛋白 E 小鼠进行苏木精-伊红（HE）染色来评估动脉粥样硬化病变的面积和大小。同时，使用氧化型低密度脂蛋白（oxLDL）在三种不同类型的细胞中模拟动脉粥样硬化。然后，用恩格列净处理巨噬细胞（RAW264.7）、人主动脉平滑肌细胞（HASMCs）和人脐静脉内皮细胞（HUVECs）后，用 Western blot 测定自噬相关蛋白和促炎细胞因子的水平，并通过共聚焦显微镜检测绿色荧光蛋白（GFP）-微管相关蛋白轻链 3（LC3）斑点，以确认自噬体的形成。油红 O 染色检测巨噬细胞和 HASMCs 的泡沫化，流式细胞术用于细胞周期分析。5-乙炔基-2'-脱氧尿苷（EdU）、细胞计数试剂盒-8（CCK-8）和划痕实验也用于检查 HASMCs 的增殖和迁移。恩格列净抑制 ApoE 小鼠动脉粥样硬化病变的进展。恩格列净还通过腺苷酸活化蛋白激酶（AMPK）信号通路诱导 RAW246.7 细胞、HASMCs 和 HUVECs 中的自噬，并显著增加 Beclin1 蛋白、LC3B-II/I 比值和 p-AMPK 蛋白的水平。此外，恩格列净通过诱导自噬降低了 P62 和促炎细胞因子的蛋白水平，并抑制了 RAW246.7 细胞和 HASMCs 的泡沫化以及炎症因子的表达。恩格列净通过 AMPK 信号通路激活自噬，从而延缓动脉粥样硬化的进展。此外，流式细胞术、EdU 实验、CCK-8 细胞活力实验和划痕实验的结果表明，恩格列净阻断了 HASMCs 的增殖和迁移。恩格列净通过 AMPK 信号通路激活自噬，从而延缓动脉粥样硬化的演变，这表明它可能成为治疗动脉粥样硬化的一种新的有效药物。
