Lu Zhiyao, Zhou Xuqian, Liu Xiaowen, Liu Chunyan, Zhang Junfeng, Dong Lei
State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, 163 Xianlin Avenue, Nanjing 210023, China.
Wuxi Xishan NJU Institute of Applied Biotechnology, Wuxi 214000, China.
Int J Mol Sci. 2025 Jul 25;26(15):7183. doi: 10.3390/ijms26157183.
The development of functional endothelial monolayers on synthetic vascular grafts remains challenging, particularly for small-diameter vessels (<6 mm) prone to thrombosis. Here, we present a pharmacological strategy combining 8-(4-chlorophenylthio) adenosine 3',5'-cyclic monophosphate sodium salt (pCPT-cAMP, a tight junction promoter) with nitric oxide/cGMP pathway agonists 3-morpholinosydnonimine (SIN-1), captopril, and sildenafil) to enhance endothelialization. In human umbilical vein endothelial cells (HUVECs), this four-agent cocktail induced a flat, extended phenotype with a 3-fold increased cell area and 57.5% fewer cells required for surface coverage compared to controls. Immunofluorescence analysis revealed enhanced ZO-1 expression and continuous tight junction formation, while sustained nitric oxide (NO) production (3.9-fold increase) and restored prostacyclin (PGI) secretion demonstrated preserved endothelial functionality. Anticoagulation assays confirmed a significant reduction in thrombus formation ( < 0.01) via dual inhibition of platelet activation and thrombin binding. These findings establish a synergistic drug combination that promotes rapid endothelialization while maintaining antithrombogenic activity, offering a promising solution for small-diameter vascular grafts. Further studies should validate long-term stability and translational potential in preclinical models.
在合成血管移植物上构建功能性内皮单层仍然具有挑战性，特别是对于易于形成血栓的小直径血管（<6mm）而言。在此，我们提出一种药理学策略，即将8-（4-氯苯硫基）腺苷3'，5'-环磷酸钠盐（pCPT-cAMP，一种紧密连接促进剂）与一氧化氮/cGMP途径激动剂3-吗啉代 sydnonimine（SIN-1）、卡托普利和西地那非联合使用，以增强内皮化。在人脐静脉内皮细胞（HUVECs）中，与对照组相比，这种四药组合诱导出一种扁平、伸展的表型，细胞面积增加了3倍，表面覆盖所需的细胞数量减少了57.5%。免疫荧光分析显示ZO-1表达增强且紧密连接持续形成，而持续的一氧化氮（NO）产生（增加3.9倍）和恢复的前列环素（PGI）分泌表明内皮功能得以保留。抗凝试验证实，通过双重抑制血小板活化和凝血酶结合，血栓形成显著减少（<0.01）。这些发现确立了一种协同药物组合，该组合可促进快速内皮化，同时保持抗血栓形成活性，为小直径血管移植物提供了一种有前景的解决方案。进一步的研究应在临床前模型中验证其长期稳定性和转化潜力。
