胞外5′-核苷酸酶基因敲除加重小鼠静脉移植血管的血管重塑和炎症反应

doi:10.3969/j.issn.1006-7795.2025.03.016

摘要/Abstract

摘要： 目的通过观察胞外5′-核苷酸酶（ecto-5′-nucleotidase，Nt5e/CD73)基因敲除小鼠在静脉移植后再狭窄血管中的表型，为临床冠状动脉搭桥术后血管再狭窄的早期诊断和药物治疗提供潜在靶点。方法将8~10周的CD73基因敲除小鼠作为实验组，同窝野生小鼠作为对照组。取小鼠下腔静脉作为供体，采用套管法移植至同种异体小鼠的右颈动脉，建立小鼠下腔静脉－颈动脉血管移植模型。模型第4周观察移植血管的通畅情况、血管内膜形成、弹力蛋白层的形态及炎症因子的表达。体外分离小鼠下腔静脉血管平滑肌细胞（vascular smooth muscle cells, VSMCs），利用CD73抑制剂腺苷-5′-(α,β-亚甲基)二磷酸［adenosine 5′-(alpha, beta-methylene) diphosphate, APCP］，观察VSMC的迁移和增殖的情况。结果与野生型小鼠移植静脉相比，CD73基因敲除小鼠移植静脉的血管新生内膜形成受损，弹力纤维层断裂消失，中膜细胞增殖增加，血管壁弹性下降，血流阻力增加。免疫组化结果显示，与野生型小鼠移植静脉相比，CD73基因敲除小鼠移植静脉组织中大量Mac-2阳性单核巨噬细胞浸润，细胞因子白细胞介素-1β（interleukin-1β，IL-1β）、IL-6、血管转化生长因子β（transforming growth factor β，TGF-β）的表达明显增加，加重静脉组织中的炎症反应和血管重塑。VSMCs的划痕实验和细胞增殖实验结果显示，抑制CD73促进VSMCs的增殖，损伤VSMCs的迁移功能。结论 CD73基因敲除加重静脉移植血管的血管重塑和炎症反应，对临床上拟作冠状动脉血管搭桥术的CD73基因缺陷的患者可提供精准诊断和治疗。

关键词: Nt5e基因, 静脉移植, 血管再狭窄, 血管重塑, 平滑肌细胞

Abstract: Objective To examine the phenotypic characteristics of Ecto-5′-nucleotidase (Nt5e/CD73) gene knockout mice in restenosis of blood vessels after vein transplantation so as to identify potential targets for early diagnosis and drug treatment of vascular restenosis after clinical coronary artery bypass surgery. Methods CD73 gene knockout mice aged 8-10 weeks were used as the experimental group, and the littermate wild-type mice were used as the control group. Using the inferior vena cava of mice as a donor, we transplanted to the right carotid artery of allogeneic mice with a trocar method to establish a mouse inferior vena cava carotid artery vascular transplantation model. At the 4th week post-model establishment, we systematically evaluated the patency of the transplanted blood vessels, the formation of the vascular intima, the proliferation of the media, the morphology of the elastin layer, and the expression of inflammatory factors. The vascular smooth muscle cells (VSMCs) from the inferior vena cava of mice were isolated in vitro, and the migration and proliferation were assessed with CD73 inhibitor adenosine 5'-(alpha, beta-methylene) diphosphate (APCP). Results In CD73 knockout mice, neointima formation was impaired, the elastic fiber layer was disrupted and lost, and medial smooth muscle cells proliferated more actively. These changes ultimately led to decreased vascular wall elasticity and increased blood flow resistance. The immunohistochemical staining results suggest that in CD73 gene knockout mice the transplanted vein tissue showed extensive infiltration of Mac-2 positive macrophages, and the expression of cytokines interleukin-1 β (IL-1 β), IL-6, and transforming growth factor β (TGF-β) was significantly increased. CD73 deficiency exacerbated inflammatory responses and vascular remodeling in venous tissues. Scratch wound healing and cell proliferation assays revealed that CD73 inhibition promoted VSMCs proliferation, yet concurrently impaired their migratory capacity. Conclusion Knockout of the CD73 gene exacerbates vascular remodeling and inflammatory response in vein grafts, offering crucial insights for the precise diagnosis and targeted treatment of patients with CD73 gene defects undergoing coronary artery bypass surgery in clinical practice.

Key words: Nt5e gene, vein graft, vascular restenosis, vascular remodeling, smooth muscle cell

中图分类号:

R654.3

刘婷婷, 石洪涛, 徐慧, 杜杰, 朴春梅. 胞外5′-核苷酸酶基因敲除加重小鼠静脉移植血管的血管重塑和炎症反应[J]. 首都医科大学学报, 2025, 46(3): 511-519.

Liu Tingting, Shi Hongtao, Xu Hui, Du jie, Piao Chunmei. Ecto-5′-nucleotidase (Nt5e/CD73) gene knockout exacerbates vascular remodeling and inflammatory response in mice after intravenous transplantation[J]. Journal of Capital Medical University, 2025, 46(3): 511-519.

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