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

甄子怡 张嘉滢 巩泽 杨伟钢 李琦 杨筱瑶 陈畅

doi:10.16098/j.issn.0529-1356.2026.01.013

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Acta Anatomica Sinica ›› 2026, Vol. 57 ›› Issue (1) : 84-91. DOI: 10.16098/j.issn.0529-1356.2026.01.013

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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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.

Key words

Intimal hyperplasia / Leflunomide / Endothelial cell / Cell migration / Reactive oxygen species / Western-blotting / Rat

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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

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