College of Pharmacy, Anhui University of Chinese Medicine, Hefei, 230012, China.
Anhui Key Laboratory for Research and Development of Traditional Chinese Medicine, Hefei, 230012, China.
Curr Neurovasc Res. 2023;20(1):76-84. doi: 10.2174/1567202620666230202124037.
Paeonol (Pae), the main active compound of the root of Paeonia albiflora, is efficacious in treating atherosclerosis (AS). Endothelial dysfunction is throughout the pathological progression of AS. It is expected that inhibition of Endothelial-to-mesenchymal transition (EndMT) will be a key target for AS treatment.
In this study, we investigated the molecular mechanism of the regulatory effect of Pae on EndMT in human umbilical vein endothelial cells (HUVECs).
Cell cytotoxicity, proliferation, and migration were detected by CCK-8, the wound healing assay, and EdU staining, respectively. The protein expressions were measured by Western blot or immunofluorescence staining. Immunofluorescence staining was performed to indicate endothelial cells undergoing EndMT in ApoE mice. TGF-β1-induced EndMT assays were performed in HUVECs and the effect of Pae was explored.
We demonstrated that Pae could improve induced TGF-β1-EndMT and . Mechanism study revealed that Pae directly bonds to the activin-like kinase 5 (ALK5, also known as TGFβ type I receptor), inhibited downstream Smad2/3 phosphorylation, and thus alleviated EndMT. Notably, overexpression of ALK5 significantly reversed the inhibitory effect of Pae on EndMT in HUVECs.
Our results indicate that ALK5 is a promising druggable target for AS, and pharmacological regulation of ALK5-Smad2/3 signaling pathway with small-molecule holds great potential to benefit AS patients.
丹皮酚（Pae）是白芍根的主要活性化合物，在治疗动脉粥样硬化（AS）方面具有疗效。内皮功能障碍贯穿 AS 的病理进展过程。预计抑制内皮-间充质转化（EndMT）将成为 AS 治疗的关键靶点。
本研究旨在探讨丹皮酚对人脐静脉内皮细胞（HUVEC）中 EndMT 的调节作用的分子机制。
通过 CCK-8 法、划痕愈合试验和 EdU 染色分别检测细胞毒性、增殖和迁移。通过 Western blot 或免疫荧光染色法测量蛋白质表达。通过免疫荧光染色法观察 ApoE 小鼠内皮细胞发生 EndMT 的情况。在 HUVEC 中进行 TGF-β1 诱导的 EndMT 试验，探讨丹皮酚的作用。
我们证明丹皮酚可以改善诱导的 TGF-β1-EndMT。机制研究表明，丹皮酚直接与激活素样激酶 5（ALK5，也称为 TGFβ 型 I 受体）结合，抑制下游 Smad2/3 磷酸化，从而减轻 EndMT。值得注意的是，ALK5 的过表达显著逆转了丹皮酚对 HUVECs 中 EndMT 的抑制作用。
我们的结果表明，ALK5 是 AS 有前途的可成药靶点，用小分子对 ALK5-Smad2/3 信号通路进行药理学调节具有很大的潜力，可使 AS 患者受益。
