Li Yi-Qi, Li Ye-Li, Li Xiao-Tong, Lv Jun-Yuan, Gao Yang, Li Wen-Na, Gong Qi-Hai, Yang Dan-Li
Joint International Research Laboratory of Ethnomedicine of Ministry of Education, Key Laboratory of Basic Pharmacology of the Ministry of Education, The Key Laboratory of Basic Pharmacology of Guizhou Province, Department of Pharmacology, School of Pharmacy, Zunyi Medical University, Zunyi, China.
Department of Pharmacology, Zhuhai Campus of Zunyi Medical University, Zhuhai, China.
Front Physiol. 2021 Jan 26;11:514494. doi: 10.3389/fphys.2020.514494. eCollection 2020.
Percutaneous coronary intervention (PCI) is the most widely used therapy for treating ischemic heart disease. However, intimal hyperplasia and restenosis usually occur within months after angioplasty. Modern pharmacological researchers have proven that osthole, the major active coumarin of (L.) Cusson, exerts potent antiproliferative effects in lung cancer cells, the human laryngeal cancer cell line RK33 and TE671 medulloblastoma cells, and its mechanism of action is related to cell cycle arrest. The goal of the present study was to observe the effect of osthole on vascular smooth muscle cell (VSMC) proliferation using platelet-derived growth factor-BB (PDGF-BB)-stimulated VSMCs isolated from rats and vascular balloon injury as models to further elucidate the molecular mechanisms underlying this activity. We detected the relative number of VSMCs by the MTT assay and EdU staining and examined cell cycle progression by flow cytometry. To more deeply probe the mechanisms, the protein expression levels of PCNA, the cyclin D1/CDK4 complex and the cyclin E1/CDK2 complex in balloon-treated rat carotid arteries and the mRNA and protein expression levels of the cyclin D1/CDK4 and cyclin E1/CDK2 complexes in VSMCs were detected by real-time RT-PCR and western blotting. The data showed that osthole significantly inhibited the proliferation of VSMCs induced by PDGF-BB. Furthermore, osthole caused apparent VSMC cycle arrest early in G0/G1 phase and decreased the expression of cyclin D1/CDK4 and cyclin E1/CDK2. Our results demonstrate that osthole can significantly inhibit PDGF-BB-induced VSMC proliferation and that its regulatory effects on cell cycle progression and proliferation may be related to the downregulation of cyclin D1/CDK4 and cyclin E1/CDK2 expression as well as the prevention of cell cycle progression from G0/G1 phase to S phase. The abovementioned mechanism may be responsible for the alleviation of neointimal hyperplasia in balloon-induced arterial wall injury by osthole.
经皮冠状动脉介入治疗（PCI）是治疗缺血性心脏病应用最广泛的疗法。然而，内膜增生和再狭窄通常会在血管成形术后数月内发生。现代药理研究人员已证实，蛇床子素（蛇床的主要活性香豆素）对肺癌细胞、人喉癌细胞系RK33和TE671髓母细胞瘤细胞具有强大的抗增殖作用，其作用机制与细胞周期阻滞有关。本研究的目的是使用从大鼠分离的血小板衍生生长因子-BB（PDGF-BB）刺激的血管平滑肌细胞（VSMC）和血管球囊损伤作为模型，观察蛇床子素对VSMC增殖的影响，以进一步阐明其作用的分子机制。我们通过MTT法和EdU染色检测VSMC的相对数量，并通过流式细胞术检测细胞周期进程。为了更深入地探究机制，通过实时RT-PCR和蛋白质印迹法检测球囊处理的大鼠颈动脉中PCNA、细胞周期蛋白D1/细胞周期蛋白依赖性激酶4（CDK4）复合物和细胞周期蛋白E1/细胞周期蛋白依赖性激酶2（CDK2）复合物的蛋白质表达水平，以及VSMC中细胞周期蛋白D1/CDK4和细胞周期蛋白E1/CDK2复合物的mRNA和蛋白质表达水平。数据显示，蛇床子素显著抑制PDGF-BB诱导的VSMC增殖。此外，蛇床子素使VSMC在G0/G1期早期明显停滞，并降低细胞周期蛋白D1/CDK4和细胞周期蛋白E1/CDK2的表达。我们的结果表明，蛇床子素可显著抑制PDGF-BB诱导的VSMC增殖，其对细胞周期进程和增殖的调节作用可能与细胞周期蛋白D1/CDK4和细胞周期蛋白E1/CDK2表达的下调以及阻止细胞周期从G0/G1期进入S期有关。上述机制可能是蛇床子素减轻球囊诱导的动脉壁损伤中新生内膜增生的原因。
