来氟米特经超氧化物歧化酶1/活性氧通路调控内皮细胞纤维性肌动蛋白生成抑制血管内膜增生

ZHEN  Ziyi1; ZHANG  Jiaying1; GONG  Ze1; YANG  Weigang1; LI  Qi2; YANG  Xiaoyao3*; CHEN  Chang1*

doi:10.16098/j.issn.0529-1356.2026.01.013

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解剖学报 ›› 2026, Vol. 57 ›› Issue (1) : 84-91. DOI: 10.16098/j.issn.0529-1356.2026.01.013

细胞和分子生物学

来氟米特经超氧化物歧化酶1/活性氧通路调控内皮细胞纤维性肌动蛋白生成抑制血管内膜增生

甄子怡¹张嘉滢¹巩泽¹杨伟钢¹李琦²杨筱瑶^3*陈畅^1*

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Leflunomide inhibiting vascular intimal hyperplasia by regulating endothelial cell fibrous actin generation via superoxide dismutase 1/reactive oxygen species pathway

ZHEN Zi-yi¹, ZHANG Jia-ying¹, GONG Ze¹, YANG Wei-gang¹, LI Qi², YANG Xiao-yao^3*, CHEN Chang^1*

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摘要

目的探讨来氟米特（Lef）促进损伤血管内皮细胞迁移修复的作用。方法雄性SD大鼠随机分为4组：正常组、正常给药组、内膜增生模型组和内膜增生模型给药组。应用2F球囊导管机械性损伤SD大鼠左颈总动脉建立血管内膜增生模型，正常给药组和内膜增生模型给药组腹腔连续14d注射来氟米特（5 mg/kg）。HE染色观察血管结构，免疫荧光和Real-time PCR检测血小板-内皮细胞黏附分子1（PECAM-1/CD31）表达。体外培养大鼠主动脉内皮细胞（RAECs），然后以Lef，H₂O₂及超氧化物歧化酶1小干扰RNA（SOD-1siRNA）等处理细胞。CCK-8检测RAECs活力，5-乙炔基-2’-脱氧尿苷（EdU）检测细胞增殖。二氢乙锭（DHE）染色检测RAECs中活性氧（ROS）生成。Western blotting检测RAECs中SOD-1的表达。Transwell检测RAECs迁移。免疫荧光检测RAECs骨架蛋白纤维性肌动蛋白（F-actin）的表达。结果与内膜增生模型组相比，来氟米特可明显抑制由血管机械性损伤所引起的血管内膜增生，促进损伤血管CD31的表达。来氟米特拮抗由H₂O₂引起的RAECs的迁移抑制和F-actin生成抑制。RAECs转染SOD-1siRNA实验发现，来氟米特通过SOD-1调控由H₂O₂所引起的细胞迁移抑制，拮抗由H₂O₂引起的RAECs内ROS的生成。结论来氟米特通过SOD-1/ROS通路调控损伤血管内皮细胞的骨架蛋白F-actin生成，从而抑制血管内膜增生。

Abstract

Objective To investigate the role of leflunomide (Lef) in promoting the migration and repair of damaged rat aortic endothelial cells (RAECs).Methods Male SD rats were randomly divided into 4 groups: normal group, normal administration group (leflunomide, intraperitoneal injection 5 mg/kg for 14 days), intimal hyperplasia model group and intimal hyperplasia model administration group (leflunomide, 5 mg/kg for 14 days). HE staining was used to observe vascular architecture; immunofluorescence staining and real-time PCR were used to detect platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31) expression. After isolating and culturing RAECs in vitro, the cells were incubated with leflunomide, H₂O₂ and superoxide dismutase 1 small interfering RNA (SOD-1 siRNA); CCK-8 and 5-ethynyl-2′-deoxyuridine (EdU) were used to detect RAECs viability and proliferation; dihydroethidium (DHE) staining was used to detect reactive oxygen species (ROS); Western blotting was used to detect the expression of SOD-1 in RAECs. Transwell assays were used for cell migration; immunofluorescence staining was used to detect fibrous actin (F-actin).Results Compared to the model group, leflunomide administration significantly inhibited mechanical vascular injury-induced intimal hyperplasia; leflunomide promoted CD31 expression in injured vessels; leflunomide counteracted H₂O₂-induced inhibition of RAECs migration; leflunomide antagonized H₂O₂-induced ROS generation in RAECs via SOD-1; leflunomide reversed H₂O₂-mediated suppression of F-actin cytoskeletal protein formation in RAECs.Conclusion Leflunomide inhibits vascular intimal hyperplasia by regulating the formation of F-actin in the cytoskeleton of injured vascular endothelial cells through the SOD-1/ROS pathway.

导出引用

甄子怡张嘉滢巩泽杨伟钢李琦杨筱瑶陈畅. 来氟米特经超氧化物歧化酶1/活性氧通路调控内皮细胞纤维性肌动蛋白生成抑制血管内膜增生[J]. 解剖学报. 2026, 57(1): 84-91 https://doi.org/10.16098/j.issn.0529-1356.2026.01.013

ZHEN Zi-yi, ZHANG Jia-ying, GONG Ze, YANG Wei-gang, LI Qi, YANG Xiao-yao, CHEN Chang.

Leflunomide inhibiting vascular intimal hyperplasia by regulating endothelial cell fibrous actin generation via superoxide dismutase 1/reactive oxygen species pathway

[J]. Acta Anatomica Sinica. 2026, 57(1): 84-91 https://doi.org/10.16098/j.issn.0529-1356.2026.01.013

中图分类号： R322.1 R96

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