Department of Cardiology, the First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
Eur Rev Med Pharmacol Sci. 2020 Jan;24(2):905-914. doi: 10.26355/eurrev_202001_20075.
The aim of this study was to explore the association between the expression of adenosine monophosphate-activated protein kinase (AMPK) pathway and adiponectin (APN), leptin, and vascular endothelial function in rats with coronary heart disease (CHD).
Experimental rats were divided into three groups, including: control (Col) group, CHD model (CHD) group, and CHD+AMPK activator (CHD+AICAR) group. Except those in Col group, all rats were fed with high-fat diet and intraperitoneally injected with pituitrin to establish the CHD model. The levels of serum APN, leptin, and endothelin-1 (ET-1) were determined via enzyme-linked immunosorbent assay (ELISA). The content of serum nitric oxide (NO) was detected using the nitrate reductase method. Meanwhile, the expression of AMPK pathway-related protein AMPKα in vascular endothelial tissues was detected via Western blotting (WB). Aortic vascular endothelial cells (VECs) were cultured with AICAR or ET-1 in vitro. Subsequently, the expressions of AMPK pathway and protein kinase B (AKT) pathway-related proteins were determined through co-immunoprecipitation and WB. Moreover, the expression level of NO in VECs was determined using the DAF-FM DA fluorescence probe.
Compared with Col group, CHD group showed significantly decreased levels of serum APN and NO (p<0.05), significantly increased the levels of leptin and ET-1 (p<0.05), as well as remarkably decreased protein expression of p-AMPKα in vascular endothelial tissues (p<0.05). After injection of AMPK activator AICAR (200 mg/kg), the protein expression of p-AMPKα in CHD rats was significantly activated (p<0.05). The levels of serum APN and NO were remarkably upregulated (p<0.05), while the levels of leptin and ET-1 were significantly reduced (p<0.05). Besides, AICAR could evidently activate the activity of AMPK pathway in VECs in vitro, upregulate the protein levels of p-eNOS (Ser1177) and p-AMPKα, and promote the secretion of NO (p<0.05). In addition, AICAR remarkably inhibited ET-1-induced expression of AKT pathway (p<0.05).
Activating the AMPK pathway may play a positive role in the normal function of VECs and exert a certain curative effect on CHD in rats.
本研究旨在探讨腺苷酸活化蛋白激酶（AMPK）通路与冠心病（CHD）大鼠脂联素（APN）、瘦素和血管内皮功能之间的关系。
实验大鼠分为三组：对照组（Col）、CHD 模型组（CHD）和 CHD+AMPK 激活剂（CHD+AICAR）组。除 Col 组外，其余大鼠均给予高脂饮食喂养，并腹腔注射垂体后叶素建立 CHD 模型。采用酶联免疫吸附法（ELISA）测定血清 APN、瘦素和内皮素-1（ET-1）水平。硝酸还原酶法检测血清一氧化氮（NO）含量。同时，采用 Western blot（WB）法检测血管内皮组织中 AMPK 通路相关蛋白 AMPKα的表达。体外采用 AICAR 或 ET-1 培养大鼠主动脉血管内皮细胞（VECs）。随后，通过免疫共沉淀和 WB 法检测 AMPK 通路和蛋白激酶 B（AKT）通路相关蛋白的表达。此外，采用 DAF-FM DA 荧光探针检测 VECs 中 NO 的表达水平。
与 Col 组相比，CHD 组大鼠血清 APN 和 NO 水平显著降低（p<0.05），瘦素和 ET-1 水平显著升高（p<0.05），血管内皮组织中 p-AMPKα蛋白表达显著降低（p<0.05）。给予 200mg/kg AMPK 激活剂 AICAR 后，CHD 大鼠血管内皮组织中 p-AMPKα蛋白表达显著激活（p<0.05）。血清 APN 和 NO 水平显著升高（p<0.05），瘦素和 ET-1 水平显著降低（p<0.05）。此外，AICAR 可明显激活体外 VECs 中 AMPK 通路的活性，上调 p-eNOS（Ser1177）和 p-AMPKα蛋白水平，并促进 NO 的分泌（p<0.05）。另外，AICAR 显著抑制 ET-1 诱导的 AKT 通路表达（p<0.05）。
激活 AMPK 通路可能对 VECs 的正常功能发挥积极作用，并对大鼠 CHD 具有一定的治疗作用。
