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Effect of Rutin on the ox-LDL-Induced Vascular Endothelial Cell Injury by Regulating NEXN-AS1/miR-410-3p Pathway

Effect of Rutin on the ox-LDL-Induced Vascular Endothelial Cell Injury by Regulating NEXN-AS1/miR-410-3p Pathway

Feng Xu, Weijing Fan and Guobin Liu*

Department of Peripheral Vascular Diseases, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, 200000, China

 
* Corresponding author: liuguobin1964@163.com

ABSTRACT

This study aims to investigate the effect of rutin on ox-LDL-induced vascular endothelial cell injury, and to analyze whether the mechanism is related to the regulation ofNEXN-AS1 and miR-410-3p expression. The CCK-8 method was used to detect the toxicity of rutin of different concentrations (0, 25, 50, 100, 200, 400 μmol/L) on human umbilical vein endothelial cells (HUVECs). HUVECs were treated with 100 mg/L ox-LDL for 24 h for establishing HUVECs injury model, and then treated with rutin in different doses. The cell activity and apoptosis rate of HUVECs were analyzed by CCK-8 method and flow cytometry, respectively. The levels of intracellular catalase (CAT), superoxide dismutase (SOD) and malondialdehyde (MDA) were measured using commercial kit, and expression levels ofNEXN-AS1 and miR-410-3p were detected using real-time quantitative PCR analysis. The dual luciferase experiment was applied to verify the targeted relationship betweenNEXN-AS1 and miR-410-3p. HUVECs were transfected with NEXN-AS1 overexpression vector, and then treated with ox-LDL for 24 h before detecting the activity, apoptosis rate and oxidative stress level of HUVECs. The low concentrations (25, 50, 100 μmol/L) of rutin had no toxic effect on HUVECs, and the activity of HUVECs was significantly reduced after intervention with high concentrations (200, 400 μmol/L) of rutin. ox-LDL treatment significantly reduced HUVECs activity, CAT and SOD levels,NEXN-AS1 expression (P<0.05), and increased apoptosis rate, MDA level, and miR-410-3p expression (P<0.05). Rutin (25, 50, 100 μmol/L) treatment reduced ox-LDL-induced apoptosis and MDA levels (P<0.05), increased cell viability, CAT and SOD levels (P<0.05), and up-regulated NEXN-AS1 Expression (P<0.05), down-regulated miR-410-3p expression (P<0.05). miR-410-3p is the target gene ofNEXN-AS1. Overexpression ofNEXN-AS1 could reduce ox-LDL-induced apoptosis and MDA levels (P<0.05), increase cell viability, CAT and SOD levels (P<0.05), and down-regulate miR-410-3p expression (P<0.05). Interference withNEXN-AS1 could reverse the effects of rutin on ox-LDL-induced proliferation, apoptosis and oxidative stress of vascular endothelial cells. It is concluded that rutin had a protective effect on ox-LDL-induced vascular endothelial apoptosis and oxidative injury, and its mechanism may be related to the up-regulation ofNEXN-AS1/miR-410-3p pathway.
 

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Pakistan Journal of Zoology

August

Vol. 54, Iss. 4, Pages 1501-2001

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