circSEC24A靶向miR-152-3p调控ox-LDL诱导的人脑微血管内皮细胞损伤

doi:10.19854/j.cnki.1008-2425.2022.02.0008

立体定向和功能性神经外科杂志 ›› 2022, Vol. 35 ›› Issue (2): 100-106.DOI: 10.19854/j.cnki.1008-2425.2022.02.0008

circSEC24A靶向miR-152-3p调控ox-LDL诱导的人脑微血管内皮细胞损伤

黄财城, 陈媚丽, 黄国堡, 陈菲

352100 宁德宁德师范学院附属宁德市医院神经内科,宁德师范学院附属宁德市医院肿瘤外科

收稿日期:2022-03-10 出版日期:2022-04-25 发布日期:2022-07-14
通讯作者: 陈菲 5472002@qq.com
基金资助:
宁德市科学技术计划项目(编号:20190069)

circSEC24A regulats ox-LDL-induced human brain microvascular endothelial cell damage by targeting miR-152-3p

Huang Caicheng, Chen Meili, Huang Guobao, et al

1. Department of Neurology,Ningde City Hospital Affiliated to Ningde Normal University,Ningde 352100,Fujian.
2. Department of oncology,Ningde City Hospital Affiliated to Ningde Normal University,Ningde,352100,China

Received:2022-03-10 Online:2022-04-25 Published:2022-07-14

摘要/Abstract

摘要： 目的探讨circSEC24A对氧化型低密度脂蛋白(ox-LDL)诱导的人脑微血管内皮细胞损伤的影响及其可能作用机制。方法用ox-LDL诱导人脑微血管内皮细胞HBMEC建立细胞氧化损伤模型,用脂质体转染法将si-NC、si-circSEC24A、miR-NC、miR-152-3p mimics、si-circSEC24A与anti-miR-NC、si-circSEC24A与anti-miR-152-3p分别转染至HBMEC后用ox-LDL处理24 h;qRT-PCR法检测circSEC24A和miR-152-3p的表达量;利用试剂盒检测MDA的水平与SOD、CAT的活性;流式细胞术检测细胞凋亡率;双荧光素酶报告实验验证circSEC24A与miR-152-3p的靶向调控作用。Western blot法检测cleaved-caspase3、cleaved-caspase9蛋白表达量。结果 ox-LDL诱导的HBMEC中circSEC24A的表达量升高(P＜0.05),MDA的水平、细胞凋亡率和cleaved-caspase3、cleaved-caspase9蛋白水平升高(P＜0.05),miR-152-3p的表达量降低(P＜0.05),SOD、CAT的活性降低(P＜0.05);转染si-circSEC24A或转染miR-152-3p mimics均可降低ox-LDL诱导的HBMEC中MDA的水平(P＜0.05),并可降低细胞凋亡率和cleaved-caspase3、cleaved-caspase9蛋白水平(P＜0.05),而SOD、CAT的活性升高(P＜0.05);circSEC24A靶向调控miR-152-3p的表达;共转染si-circSEC24A与anti-miR-152-3p后MDA的水平升高(P＜0.05),细胞凋亡率和cleaved-caspase3、cleaved-caspase9蛋白水平升高(P＜0.05),而SOD、CAT的活性降低(P＜0.05)。结论干扰circSEC24A表达可靶向调控miR-152-3p表达而抑制细胞氧化应激及凋亡从而减轻ox-LDL诱导的人脑微血管内皮细胞损伤。

关键词: 人脑微血管内皮细胞, 氧化型低密度脂蛋白, circSEC24A, miR-152-3p, 氧化应激, 凋亡

Abstract: Objective To explore the effect of circSEC24A on human brain microvascular endothelial cell injury induced by ox-LDL and its possible mechanism. Methods Human brain microvascular endothelial cells HBMEC were induced with ox-LDL to establish a cellular oxidative damage model,and si-NC,si-circSEC24A,miR-NC,miR-152-3p mimics,si-circSEC24A and anti-miR-NC,si-circSEC24A and anti-miR-152-3p were transfected into HBMEC and treated with ox-LDL for 24 h.qRT-PCR method was used to detect the expression levels of circSEC24A and miR-152-3p.The level of MDA and the activity of SOD and CAT were tested using kits.Flow cytometry was used to detect the rate of apoptosis.The targeted regulation of circSEC24A and miR-152-3p was verified by dual luciferase report assay.Western blot was used to detect the protein expression of cleaved-caspase3 and cleaved-caspase9. Results The expression of circSEC24A in ox-LDL-induced HBMEC was increased (P＜0.05),the level of MDA,the apoptosis rate and the protein levels of cleaved-caspase3,cleaved-caspase9 were increased (P＜0.05),while the expression of miR-152-3p was decreased (P＜0.05),and the activities of SOD,CAT were decreased (P＜0.05)..Transfection of si-circSEC24A or miR-152-3p mimics could reduce the level of MDA in HBMEC induced by ox-LDL (P＜0.05),and could reduce the apoptosis rate and the protein levels of cleaved-caspase3,cleaved-caspase9 (P＜0.05),while the activities of SOD and CAT were increased (P＜0.05).circSEC24A could target and regulate the expression of miR-152-3p.After co-transfection of si-circSEC24A and anti-miR-152-3p,the level of MDA was increased (P＜0.05),the apoptosis rate and the protein levels of cleaved-caspase3,cleaved-caspase9 were increased (P＜0.05),while the activities of SOD and CAT were decreased (P＜0.05). Conclusion Interfering with circSEC24A could inhibit cell oxidative stress and apoptosis by targeting miR-152-3p,thereby reducing ox-LDL-induced human brain microvascular endothelial cells damage.

Key words: Human brain microvascular endothelial cells, Xxidized low-density lipoprotein, CircSEC24A, MiR-152-3p, Oxidative stress, Apoptosis

中图分类号:

R743 R363

黄财城, 陈媚丽, 黄国堡, 陈菲. circSEC24A靶向miR-152-3p调控ox-LDL诱导的人脑微血管内皮细胞损伤[J]. 立体定向和功能性神经外科杂志, 2022, 35(2): 100-106.

Huang Caicheng, Chen Meili, Huang Guobao, et al. circSEC24A regulats ox-LDL-induced human brain microvascular endothelial cell damage by targeting miR-152-3p[J]. Chinese Journal of Stereotactic and Functional Neurosurgery, 2022, 35(2): 100-106.

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circSEC24A靶向miR-152-3p调控ox-LDL诱导的人脑微血管内皮细胞损伤

circSEC24A regulats ox-LDL-induced human brain microvascular endothelial cell damage by targeting miR-152-3p

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