Wu Xize, Pan Xue, Kang Jian, Huang Yuxi, Ren Jiaqi, Pan Jiaxiang, Yu Kaifeng, Li Yue
The First Clinical College, Liaoning University of Traditional Chinese Medicine, Shenyang, China.
College of Traditional Chinese Medicine, Dazhou Vocational College of Chinese Medicine, Dazhou, Sichuan, China.
Front Pharmacol. 2025 Mar 18;16:1524736. doi: 10.3389/fphar.2025.1524736. eCollection 2025.
Ferulic acid (FA) has shown potential in treating atherosclerosis (AS) by improving lipid metabolism and exerting anti-hypoxic effects. This study aimed to validate the mechanism of FA in AS through experiments.
Network analysis was employed to predict the mechanisms underlying the therapeutic effects of FA on AS. An foam cell model was established using RAW 264.7 cells treated with ox-LDL. Cellular lipid accumulation was detected using Oil Red O staining; cell viability was assessed by cell counting kit-8; mitochondrial morphology and function were evaluated by transmission electron microscopy and JC-1 staining; apoptosis levels were detected by TUNEL and DAPI staining; mitochondrial Fe content was measured by Mito-FerroGreen; and Western blot was performed to determine the protein expression levels of HIF-1α, Bax, Bcl2, GPX4, and EGFR.
Network analysis suggested that FA may exert its therapeutic effects on AS through the HIF-1 signaling pathway and is closely associated with the regulation of ferroptosis and apoptosis. FA upregulated the expression of ALOX5, BCL2, ERN1, GPX4, NOS3, and SLC2A1 mRNA and downregulated the expression of BAX, CYCS, EGFR, FLT1, HIF1A, NFKB1, NOS2, PARP1, and STAT3 mRNA. experiments demonstrated that FA reduces lipid accumulation, increases cell viability, improves mitochondrial function, and decreases reactive oxygen species content. Additionally, FA inhibited ferroptosis and apoptosis by suppressing the HIF-1 signaling pathway, up-regulating the expression of GPX4 and Bcl2, and down-regulating the expression of HIF-1α and Bax protein. HIF-1 agonists reversed these effects by activating the HIF-1 signaling pathway.
FA improves mitochondrial function and suppresses ferroptosis and apoptosis by inhibiting the HIF-1 signaling pathway, thereby treating AS.
阿魏酸（FA）已显示出通过改善脂质代谢和发挥抗缺氧作用来治疗动脉粥样硬化（AS）的潜力。本研究旨在通过实验验证FA在AS中的作用机制。
采用网络分析预测FA对AS治疗作用的潜在机制。使用经氧化型低密度脂蛋白（ox-LDL）处理的RAW 264.7细胞建立泡沫细胞模型。用油红O染色检测细胞脂质积累；通过细胞计数试剂盒-8评估细胞活力；用透射电子显微镜和JC-1染色评估线粒体形态和功能；用TUNEL和DAPI染色检测凋亡水平；用Mito-FerroGreen测量线粒体铁含量；并进行蛋白质免疫印迹法以确定缺氧诱导因子-1α（HIF-1α）、Bax、Bcl-2、谷胱甘肽过氧化物酶4（GPX4）和表皮生长因子受体（EGFR）的蛋白表达水平。
网络分析表明，FA可能通过HIF-1信号通路对AS发挥治疗作用，且与铁死亡和凋亡的调控密切相关。FA上调了5-脂氧合酶（ALOX5）、B细胞淋巴瘤/白血病-2（BCL2）、内质网应激反应蛋白1（ERN1）、GPX4、一氧化氮合酶3（NOS3）和溶质载体家族2成员1（SLC2A1）mRNA的表达，并下调了BAX、细胞色素c（CYCS）、EGFR、血管内皮生长因子受体1（FLT1）、HIF1A、核因子κB亚基1（NFKB1）、一氧化氮合酶2（NOS2）、聚（ADP-核糖）聚合酶1（PARP1）和信号转导和转录激活因子3（STAT3）mRNA的表达。实验表明，FA减少脂质积累，增加细胞活力，改善线粒体功能，并降低活性氧含量。此外，FA通过抑制HIF-1信号通路、上调GPX4和Bcl-2的表达以及下调HIF-1α和Bax蛋白的表达来抑制铁死亡和凋亡。HIF-1激动剂通过激活HIF-1信号通路逆转了这些作用。
FA通过抑制HIF-1信号通路改善线粒体功能并抑制铁死亡和凋亡，从而治疗AS。
