AMPK 通路表达与冠心病大鼠脂联素、瘦素和血管内皮功能的关系。
Association between expression of AMPK pathway and adiponectin, leptin, and vascular endothelial function in rats with coronary heart disease.
机构信息
Department of Cardiology, the First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
出版信息
Eur Rev Med Pharmacol Sci. 2020 Jan;24(2):905-914. doi: 10.26355/eurrev_202001_20075.
OBJECTIVE
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).
MATERIALS AND METHODS
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.
RESULTS
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).
CONCLUSIONS
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.
目的
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。
材料与方法
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。
结果
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）。
结论
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。
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