Institute of Cardiac Surgery, Department of Cardiovascular Surgery, Changhai Hospital, Naval Medical University, 168 Changhai Road, Shanghai, 200433, China.
, National Clinical Research Center of Kidney Diseases, Jinling Hospital, Nanjing University School of Medicine, Nanjing, 210018, China.
Cardiovasc Drugs Ther. 2021 Feb;35(1):103-111. doi: 10.1007/s10557-020-07089-4. Epub 2020 Oct 12.
Vein graft failure (VGF) is an important limitation for coronary artery bypass graft (CABG) surgery. Inhibition of the excessive proliferation and migration of venous smooth muscle cells (SMCs) is an effective strategy to alleviate VGF during the CABG perioperative period. In the present study, we aimed to explore the role and potential mechanism of all-trans retinoic acid (ATRA) on preventing vein grafts stenosis.
The autogenous vein grafts model was established in the right jugular artery of rabbits. Immunohistochemistry staining and western blot assays were used to detected the protein expression, while real-time PCR assay was applied for mRNAs expression detection. The interaction between proteins was identified by co-immunoprecipitation assay. The Cell Counting Kit-8 and wound-healing assays were used to investigate the role of ATRA on human umbilical vein smooth muscle cells (HUVSMCs) function. Cell cycle progression was identified by flow cytometry assay.
Vein graft stenosis and SMCs hyperproliferation were confirmed in vein grafts by histological and Ki-67 immunohistochemistry assays. Treatment of ATRA (10 mg/kg/day) significantly mitigated the stenosis extent of vein grafts, demonstrated by the decreased thickness of intima-media, and decreased Ki-67 expression. ATRA could repress the PDGF-bb-induced excessive proliferation and migration of HUVSMCs, which was mediated by Rb-E2F dependent cell cycle inhibition. Meanwhile, ATRA could reduce the interaction between KLF5 and RARα, thereby inhibiting the function of cis-elements of KLF5. KLF5-induced inducible nitric oxide synthase (iNOS) expression activation could be significantly inhibited by ATRA.
These results suggested that ATRA treatment may represent an effective prevention and therapy avenue for VGF.
静脉移植物失败（VGF）是冠状动脉旁路移植术（CABG）的一个重要限制因素。抑制静脉平滑肌细胞（SMCs）的过度增殖和迁移是减轻 CABG 围手术期 VGF 的有效策略。在本研究中，我们旨在探讨全反式视黄酸（ATRA）在预防静脉移植物狭窄中的作用及其潜在机制。
在兔右侧颈总动脉建立自体静脉移植物模型。免疫组织化学染色和 Western blot 检测用于检测蛋白质表达，实时 PCR 检测用于 mRNAs 表达检测。通过共免疫沉淀实验鉴定蛋白质相互作用。细胞计数试剂盒-8 和划痕愈合实验用于研究 ATRA 对人脐静脉平滑肌细胞（HUVSMCs）功能的作用。通过流式细胞术检测细胞周期进展。
通过组织学和 Ki-67 免疫组织化学检测证实静脉移植物存在静脉移植物狭窄和 SMCs 过度增殖。ATRA（10mg/kg/天）治疗显著减轻了静脉移植物的狭窄程度，表现为内膜-中膜厚度减小，Ki-67 表达减少。ATRA 可以抑制 PDGF-bb 诱导的 HUVSMCs 的过度增殖和迁移，这是通过 Rb-E2F 依赖性细胞周期抑制介导的。同时，ATRA 可以减少 KLF5 和 RARα 之间的相互作用，从而抑制 KLF5 的顺式元件的功能。ATRA 可以显著抑制 KLF5 诱导的诱导型一氧化氮合酶（iNOS）表达激活。
这些结果表明，ATRA 治疗可能是预防和治疗 VGF 的有效途径。
