Xu Tong, Hu Changlei, Zhang Guoyong, Xu Honglin, Chen Guanghong, Pang Mingjie, Zhang Jiayi, Hua Yue, Liu Bin, Zhou Yingchun
School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, 510515, China.
Dongguan Hospital, Southern Medical University (Dongguan People's Hospital), Dongguan, 523058, China.
J Ethnopharmacol. 2025 Jul 3;352:120244. doi: 10.1016/j.jep.2025.120244.
The mortality rate of cardiovascular diseases (CVD) has been increasing year by year. Percutaneous coronary intervention (PCI) is a major treatment for CVDs; however, the occurrence of in-stent restenosis (ISR) has severely impacted the efficacy of PCI. Currently, there are still no effective treatments for ISR. Neointimal hyperplasia (NIH) is the primary pathological change associated with ISR. The proliferation and migration of vascular smooth muscle cells are crucial for the occurrence of NIH. Therefore, finding effective methods to inhibit vascular smooth muscle cell proliferation, migration, and NIH is the key to ISR treatment. While Buyang Huanwu decoction (BYHWD), a blend of traditional Chinese medicinal herbs, has shown therapeutic potential against arterial stenosis diseases. However, its effectiveness on ISR remains unknown. Therefore, exploring how BYHWD influences these processes is crucial for a deeper understanding of its therapeutic effects.
The aim of this study is to investigate the impact of BYHWD on arterial in-stent restenosis and explore the potential involvement of transforming growth factor beta receptor 1 (TGFBR1) in this process.
The influence of BYHWD on the migration and proliferation of mouse vascular smooth muscle cells (MOVAS) and human aortic smooth muscle cells (HASMCs) was assessed through methylthiazolyldiphenyl-tetrazolium bromide (MTT), 5-ethynyl-2'-deoxyuridine (EdU), Transwell, wound healing, and Western blotting (WB) assays. In addition, the role of BYHWD was further validated using the TGFBR1-encoding adenovirus vector. In vivo, the impact of BYHWD on intimal formation and TGFBR1 activation was determined based on the adeno-associated virus-mediated TGFBR1 overexpression models and a left common carotid artery (LCCA) ligation mouse model.
BYHWD dose-dependently suppressed vascular smooth muscle cell (VSMC) migration and proliferation, which included inhibiting cyclin A2, cyclin dependent kinase 4 (CDK4), matrix metalloproteinase-9 (MMP9) expression, and increasing tissue inhibitor of metallopeptidase-1 (TIMP1) expression. Additionally, phosphorylated TGFBR1 (p-TGFBR1), phosphorylated mothers against decapentaplegic homolog 2 (p-Smad2), and phosphorylated mothers against decapentaplegic homolog 3 (p-Smad3) expression decreased after BYHWD administration, while their phosphorylation levels were partially reversed by the overexpression of TGFBR1. Similarly, the inhibition of BYHWD against VSMC migration and proliferation was partially mitigated by the overexpression of TGFBR1. Furthermore, BYHWD significantly attenuated NIH and decreased the p-TGFBR1, p-Smad2, and p-Smad3 levels in the LCCA model; however, such inhibitory effect could be partially reversed through the overexpression of TGFBR1.
This study provides the initial evidence supporting that BYHWD blocks VSMC migration, proliferation, and NIH by suppressing the TGFBR1 pathway. BYHWD is promising to become a candidate treatment strategy for ISR.
心血管疾病（CVD）的死亡率逐年上升。经皮冠状动脉介入治疗（PCI）是治疗CVD的主要方法；然而，支架内再狭窄（ISR）的发生严重影响了PCI的疗效。目前，对于ISR仍没有有效的治疗方法。新生内膜增生（NIH）是与ISR相关的主要病理变化。血管平滑肌细胞的增殖和迁移对于NIH的发生至关重要。因此，找到抑制血管平滑肌细胞增殖、迁移和NIH的有效方法是治疗ISR的关键。补阳还五汤（BYHWD）作为一种中药复方，已显示出对动脉狭窄疾病的治疗潜力。然而，其对ISR的有效性尚不清楚。因此，探索BYHWD如何影响这些过程对于深入了解其治疗效果至关重要。
本研究旨在探讨BYHWD对动脉支架内再狭窄的影响，并探索转化生长因子β受体1（TGFBR1）在此过程中的潜在作用。
通过甲基噻唑基二苯基溴化四氮唑（MTT）、5-乙炔基-2'-脱氧尿苷（EdU）、Transwell、伤口愈合和蛋白质免疫印迹（WB）分析评估BYHWD对小鼠血管平滑肌细胞（MOVAS）和人主动脉平滑肌细胞（HASMCs）迁移和增殖的影响。此外，使用编码TGFBR1的腺病毒载体进一步验证BYHWD的作用。在体内，基于腺相关病毒介导的TGFBR1过表达模型和左颈总动脉（LCCA）结扎小鼠模型，确定BYHWD对内膜形成和TGFBR1激活的影响。
BYHWD剂量依赖性地抑制血管平滑肌细胞（VSMC）的迁移和增殖，包括抑制细胞周期蛋白A2、细胞周期蛋白依赖性激酶4（CDK4）、基质金属蛋白酶-9（MMP9）的表达，并增加金属蛋白酶组织抑制剂-1（TIMP1）的表达。此外，给予BYHWD后，磷酸化的TGFBR1（p-TGFBR1）、磷酸化的母亲对脱磷酸化同源物2（p-Smad2）和磷酸化的母亲对脱磷酸化同源物3（p-Smad3）的表达降低，而TGFBR1的过表达部分逆转了它们的磷酸化水平。同样，TGFBR1的过表达部分减轻了BYHWD对VSMC迁移和增殖的抑制作用。此外，BYHWD显著减轻了LCCA模型中的NIH，并降低了p-TGFBR1、p-Smad2和p-Smad3的水平；然而，这种抑制作用可通过TGFBR1的过表达部分逆转。
本研究提供了初步证据支持BYHWD通过抑制TGFBR1途径阻断VSMC迁移、增殖和NIH。BYHWD有望成为ISR的候选治疗策略。
