Xie Shishun, Zhao Jianjun, Zhang Fan, Li Xiangjun, Yu Xiaoyan, Shu Zhiyun, Cheng Hongyuan, Liu Siyao, Shi Shaomin
Department of Experimental Pharmacology and Toxicology, School of Pharmaceutical Sciences, Jilin University, Changchun 130000, China; Department of Respiratory Medicine, China-Japan Union Hospital of Jilin University, Changchun 130000, China.
Department of Respiratory Medicine, China-Japan Union Hospital of Jilin University, Changchun 130000, China.
Toxicol Appl Pharmacol. 2025 Feb;495:117212. doi: 10.1016/j.taap.2024.117212. Epub 2024 Dec 22.
Abnormal proliferation and migration of pulmonary artery smooth muscle cells (PASMCs) leading to pulmonary vascular remodeling are critical factors in the development of pulmonary hypertension (pH). Dehydrodiisoeugenol (DEH), a natural phenolic compound, is renowned for its antioxidant and anti-inflammatory properties. However, the precise role and mechanisms of DEH in PH remain unclear. In this study, human PASMCs were exposed to PDGF-BB for 48 h to establish an in vitro model. Subsequently, cells were treated with DEH, and assessments of cell proliferation, migration, and apoptosis were performed using CCK-8/EdU assays, scratch/transwell assays, and flow cytometry. The results showed that PDGF-BB induced phenotypic modulation, proliferation, and migration of PASMCs while reducing apoptosis. Treatment with DEH effectively reversed these effects. Bioinformatics analysis identified mTOR as a target of DEH action. Western blot experiments were conducted to evaluate the expression of proteins involved in the mTOR/HIF1-α/HK2 signaling pathway, suggesting that DEH modulates this pathway by targeting and inhibiting mTOR. After treating cells with mTOR inhibitors, cellular glycolysis was assessed using the extracellular acidification rate (ECAR) assay. The results indicated that inhibition of mTOR phosphorylation decreased aerobic glycolysis in PASMCs and suppressed cell proliferation, migration, and apoptosis resistance, regardless of PDGF-BB treatment. Activation of mTOR reversed the inhibition of PDGF-BB-induced PASMC-related protein expression by DEH. These findings suggest that DEH inhibits aerobic glycolysis in PDGF-BB-induced PASMCs through the mTOR/HIF1-α/HK2 signaling pathway, thereby suppressing cell proliferation, migration, and resistance to apoptosis. Consequently, DEH holds promise as a novel therapeutic agent for treating pulmonary arterial hypertension.
肺动脉平滑肌细胞（PASMCs）的异常增殖和迁移导致肺血管重塑，是肺动脉高压（pH）发生发展的关键因素。脱氢二异丁香酚（DEH）是一种天然酚类化合物，以其抗氧化和抗炎特性而闻名。然而，DEH在肺动脉高压中的具体作用和机制仍不清楚。在本研究中，将人PASMCs暴露于血小板衍生生长因子BB（PDGF-BB）48小时以建立体外模型。随后，用DEH处理细胞，并使用CCK-8/EdU测定法、划痕/Transwell测定法和流式细胞术对细胞增殖、迁移和凋亡进行评估。结果表明，PDGF-BB诱导PASMCs的表型调节、增殖和迁移，同时减少凋亡。用DEH处理可有效逆转这些作用。生物信息学分析确定mTOR为DEH作用的靶点。进行蛋白质印迹实验以评估mTOR/HIF1-α/HK2信号通路中相关蛋白质的表达，表明DEH通过靶向和抑制mTOR来调节该信号通路。在用mTOR抑制剂处理细胞后，使用细胞外酸化率（ECAR）测定法评估细胞糖酵解。结果表明，抑制mTOR磷酸化可降低PASMCs中的有氧糖酵解，并抑制细胞增殖、迁移和抗凋亡能力，无论是否用PDGF-BB处理。激活mTOR可逆转DEH对PDGF-BB诱导的PASMC相关蛋白表达的抑制作用。这些发现表明，DEH通过mTOR/HIF1-α/HK2信号通路抑制PDGF-BB诱导的PASMCs中的有氧糖酵解，从而抑制细胞增殖、迁移和抗凋亡能力。因此，DEH有望成为治疗肺动脉高压的新型治疗药物。
