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S-炔丙基半胱氨酸延缓动脉粥样硬化进展并上调内皮细胞中内皮型一氧化氮合酶磷酸化

李志明¹, 李娉¹, 朱磊², 张育文³, 朱依纯¹, 王鹤³, 余波², 王铭洁^1,*

¹小分子活性物质上海高校重点实验室，上海高水平地方高校创新团队，复旦大学基础医学院生理与病理生理学系，上海200032；²复旦大学附属华山医院血管外科，上海 200040；³计算神经科学与类脑智能教育部重点实验室，复旦大学类脑智能科学与技术研究院，上海 200433

摘要

本研究旨在探究炔丙基半胱氨酸(S-propargyl-cysteine, SPRC)对延缓小鼠动脉粥样硬化进展的保护作用。用ApoE−/−小鼠通过右侧颈动脉串联双狭窄(tandem stenosis, TS)联合西方饮食构建动脉粥样硬化易损斑块小鼠模型，测定血管损伤面积、血脂和炎症水平来评估SPRC的抗动脉粥样硬化作用，组织病理学分析以评估斑块稳定性。为探究SPRC的保护机制，体外培养人脐静脉内皮细胞(human umbilical vein endothelial cells, HUVECs)并用氧化型低密度脂蛋白(oxidized low-density lipoprotein, ox-LDL)刺激，用细胞活力测定试剂盒测定细胞活力，用Western blot和RT-qPCR分别检测磷酸化内皮型一氧化氮合酶(endothelial nitric oxide synthase, eNOS)蛋白和mRNA表达。结果表明，每天80 mg/kg SPRC组小鼠主动脉弓和颈动脉病变面积显著减少，血浆胆固醇和低密度脂蛋白胆固醇水平显著降低，斑块中的胶原水平增加，且斑块中基质金属蛋白9 (matrix metalloproteinase-9, MMP-9)含量明显降低。体内实验结果表明SPRC发挥稳定斑块作用。离体实验表明100 μmol/L SPRC提高ox-LDL处理造成的细胞活力下降，提高eNOS磷酸化蛋白的表达。以上结果提示SPRC延缓了动脉粥样硬化的进展，其作用机制可能与内皮细胞中eNOS磷酸化的增加有关。

关键词： 炔丙基半胱氨酸; 动脉粥样硬化; 不稳定斑块; 人脐静脉内皮细胞; 内皮型一氧化氮合酶

S-propargyl-cysteine delays the progression of atherosclerosis and increases eNOS phosphorylation in endothelial cells 

LI Zhi-Ming¹, LI Ping¹, ZHU Lei², ZHANG Yu-Wen³, ZHU Yi-Chun¹, WANG He³, YU Bo², WANG Ming-Jie^1,*

¹Shanghai Key Laboratory of Bioactive Small Molecules, the Innovative Research Team of High-Level Local Universities in Shanghai, Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, China；²Department of Vascular Surgery, Huashan Hospital, Fudan University, Shanghai 200040, China；³Key Laboratory of Computational Neuroscience and Brain-Inspired Intelligence, Institute of Science and Technology for Brain-Inspired Intelligence, Fudan University, Shanghai 200433, China

Abstract

The present study aimed to investigate the protective effect of S-propargyl-cysteine (SPRC) on atherosclerosis progression in mice. A mouse model of vulnerable atherosclerotic plaque was created in ApoE−/− mice by carotid artery tandem stenosis (TS) combined with a Western diet. Macrophotography, lipid profiles, and inflammatory markers were measured to evaluate the antiatherosclerotic effects of SPRC compared to atorvastatin as a control. Histopathological analysis was performed to assess the plaque stability. To explore the protective mechanism of SPRC, human umbilical vein endothelial cells (HUVECs) were cultured in vitro and challenged with oxidized low-density lipoprotein (ox-LDL). Cell viability was determined with a Cell Counting Kit-8 (CCK-8). Endothelial nitric oxide synthase (eNOS) phosphorylation and mRNA expression were detected by Western blot and RT-qPCR respectively. The results showed that the lesion area quantified by en face photographs of the aortic arch and carotid artery was significantly less, plasma total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) were reduced, plaque collagen content was increased and matrix metalloproteinase-9 (MMP-9) was decreased in 80 mg/kg per day SPRC-treated mice compared with model mice. These findings support the role of SPRC in plaque stabilization. In vitro studies revealed that 100 μmol/L SPRC increased the cell viability and the phosphorylation level of eNOS after ox-LDL challenge. These results suggest that SPRC delays the progression of atherosclerosis and enhances plaque stability. The protective effect may be at least partially related to the increased phosphorylation of eNOS in endothelial cells.

Key words: S-propargyl-cysteine; atherosclerosis; unstable plaque; HUVECs; eNOS

收稿日期：　　录用日期：

通讯作者：王铭洁　　E-mail: mjwang@shmu.edu.cn

引用本文：

李志明, 李娉, 朱磊, 张育文, 朱依纯, 王鹤, 余波, 王铭洁. S-炔丙基半胱氨酸延缓动脉粥样硬化进展并上调内皮细胞中内皮型一氧化氮合酶磷酸化[J]. 生理学报 2023; 75 (3): 317-327.

LI Zhi-Ming, LI Ping, ZHU Lei, ZHANG Yu-Wen, ZHU Yi-Chun, WANG He, YU Bo, WANG Ming-Jie. S-propargyl-cysteine delays the progression of atherosclerosis and increases eNOS phosphorylation in endothelial cells . Acta Physiol Sin 2023; 75 (3): 317-327