The Biotherapy Center, Tumor Hospital of Harbin Medical University, Harbin, China.
Department of Pharmacology (State-Province Key Laboratories of Biomedicine-Pharmaceutics of China, Key Laboratory of Cardiovascular Research, Ministry of Education), College of Pharmacy, Harbin Medical University, Harbin, China.
J Vasc Res. 2024;61(3):99-108. doi: 10.1159/000532029. Epub 2023 Dec 27.
This study aimed to determine whether bone morphogenetic protein-4 (BMP-4), which increases in response to intimal hyperplasia, promotes phenotype transition in vascular smooth muscle cells (VSMCs).
Balloon injury was used to induce intimal hyperplasia in rats. Hematoxylin-eosin staining was used to detect the alteration of vascular structure. Serum levels of BMP-4 and lactate were detected by ELISA. Human aortic smooth muscle cells (HA-SMCs) were cultured. Protein and mRNA expression levels were detected through Western blot and real-time PCR. Cell migration was measured by transwell assay.
Our data showed that serum concentration of BMP-4 was upregulated after balloon injury. Treatment with BMP-4 inhibitor DMH1 (4-(6-(4-isopropoxyphenyl)pyrazolo(1,5-a)pyrimidin-3-yl)quinoline) suppressed the abnormal expression of BMP-4 and inhibited the intimal hyperplasia induced by balloon injury. Compared to BMP-4-negative medium, BMP-4-positive medium was associated with higher synthetic VSMC marker expression levels and lower in contractile gene markers in cultured HA-SMCs. Transfection of monocarboxylic acid transporters-4 (MCT-4) siRNA inhibited the excretion of lactate induced by BMP-4.
Our analyses provided evidence that BMP-4 and its regulator Smad-4 are key regulators in MCT-4-mediated lactate excretion. This indicates that BMP-4 stimulates the phenotypic transition of VSMCs via SMAD-4/MCT-4 signaling pathway.
本研究旨在确定骨形态发生蛋白-4（BMP-4）是否在血管平滑肌细胞（VSMCs）中促进表型转化，该蛋白在应对内膜增生时会增加。
采用球囊损伤法诱导大鼠内膜增生。苏木精-伊红染色检测血管结构的改变。通过 ELISA 检测血清 BMP-4 和乳酸水平。培养人主动脉平滑肌细胞（HA-SMCs）。通过 Western blot 和实时 PCR 检测蛋白和 mRNA 表达水平。通过 Transwell 测定细胞迁移。
我们的数据显示，球囊损伤后血清 BMP-4 浓度上调。BMP-4 抑制剂 DMH1（4-(6-(4-异丙氧基苯基)吡唑并(1,5-a)嘧啶-3-基)喹啉）处理抑制了异常的 BMP-4 表达，并抑制了球囊损伤引起的内膜增生。与 BMP-4 阴性培养基相比，BMP-4 阳性培养基与培养的 HA-SMCs 中更高的合成 VSMC 标志物表达水平和更低的收缩基因标志物相关。单羧酸转运蛋白-4（MCT-4）siRNA 的转染抑制了 BMP-4 诱导的乳酸排泄。
我们的分析提供了证据表明，BMP-4 及其调节因子 Smad-4 是 MCT-4 介导的乳酸排泄的关键调节剂。这表明 BMP-4 通过 SMAD-4/MCT-4 信号通路刺激 VSMCs 的表型转化。
