LncRNA FGD5-AS1靶向miR-106a-5p减轻ox-LDL诱导的HUVECs损伤

doi:10.16352/j.issn.1001-6325.2022.05.026

摘要/Abstract

摘要： 目的探讨lncRNA FGD5-AS1对氧化型低密度脂蛋白(ox-LDL)诱导的血管内皮细胞损伤的影响及其对miR-106a-5p的调控作用。方法 Ox-LDL诱导人脐静脉血管内皮细胞(HUVECs)建立细胞损伤模型,pcDNA-FGD5-AS1、anti-miR-106a-5p及其阴性对照分别转染至HUVECs后加入ox-LDL处理细胞,pcDNA-FGD5-AS1分别与miR-NC、miR-106a-5p mimics共转染至HUVECs后加入ox-LDL处理细胞;RT-qPCR检测FGD5-AS1、miR-106a-5p的表达量;试剂盒检测丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)的水平;流式细胞测量术检测细胞凋亡率;双荧光素酶报告基因实验检测FGD5-AS1与miR-106a-5p的靶向关系;Western blot检测cleaved caspase-3、cleaved caspase-9蛋白表达量。结果 Ox-LDL诱导的HUVECs中FGD5-AS1 mRNA的表达量降低(P＜0.05),而miR-106a-5p mRNA的表达量升高(P＜0.05);转染pcDNA-FGD5-AS1可降低ox-LDL诱导的HUVECs中MDA的水平、凋亡率与cleaved caspase-3、cleaved caspase-9蛋白水平(P＜0.05),而增强SOD、CAT的活性(P＜0.05);FGD5-AS1可靶向调控miR-106a-5p;转染anti-miR-106a-5p可降低ox-LDL诱导的HUVECs中MDA的水平、凋亡率与cleaved caspase-3、cleaved caspase-9蛋白水平(P＜0.05),而增强SOD、CAT的活性(P＜0.05);pcDNA-FGD5-AS1与miR-106a-5p mimics共转染后可逆转pcDNA-FGD5-AS1对ox-LDL诱导的HUVECs凋亡及氧化应激的作用。结论 FGD5-AS1过表达可通过靶向调控miR-106a-5p抑制氧化应激及凋亡,进而减轻ox-LDL诱导的血管内皮细胞损伤。

关键词: lncRNAFGD5-AS1, miR-106a-5p, 人脐静脉血管内皮细胞, 氧化应激, 凋亡

Abstract: Objective To explore the effect of lncRNA FGD5-AS1 on oxidized low-density lipoprotein (ox-LDL)-induced vascular endothelial cell damage and its regulatory effect on miR-106a-5p. Methods Ox-LDL was used to induce human umbilical vein endothelial cells(HUVECs) to establish a cellular injury model. pcDNA-FGD5-AS1, anti-miR-106a-5p and its negative control were transfected into HUVECs and then added to ox-LDL-treated cells. pcDNA-FGD5-AS1 was co-transfected with miR-NC and miR-106a-5p mimics to HUVECs, then added ox-LDL to treat cells. RT-qPCR was used to detect the expression of FGD5-AS1 and miR-106a-5p. The level of MDA, SOD, and CAT was tested with commercially available kit. Flow cytometry was used to detect the apoptosis rate. The dual luciferase reporter gene experiment was used to detect the targeting relationship between FGD5-AS1 and miR-106a-5p. Western blot was used to detect the protein expression of cleaved caspase-3 and cleaved caspase-9. Results The expression of FGD5-AS1 mRNA in HUVECs induced by ox-LDL decreased (P＜0.05) while the expression of miR-106a-5p mRNA increased (P＜0.05). Both transfection of pcDNA-FGD5-AS1 and transfection of anti-miR-106a-5p reduced the level of MDA, apoptosis rate and protein level of cleaved caspase-3, cleaved caspase-9 in HUVECs induced by ox-LDL(P＜0.05),enhanced the activity of SOD and CAT (P＜0.05). FGD5-AS1 could target at miR-106a-5p. Co-transfection of pcDNA-FGD5-AS1 and miR-106a-5p mimics could reverse the effect of pcDNA-FGD5-AS1 on ox-LDL-induced HUVECs apoptosis and oxidative stress. Conclusions Over-expression of FGD5-AS1 may inhibit oxidative stress and apoptosis through targeted regulation of miR-106a-5p, thereby reducing ox-LDL-induced vascular endothelial cell damage.

Key words: lncRNA FGD5-AS1, miR-106a-5p, human umbilical vein endothelial cells, oxidative stress, apoptosis

中图分类号:

R394

郭婧, 李亚洁, 牟寒霜. LncRNA FGD5-AS1靶向miR-106a-5p减轻ox-LDL诱导的HUVECs损伤[J]. 基础医学与临床, 2022, 42(5): 768-775.

GUO Jing, LI Ya-jie, MOU Han-shuang. LncRNA FGD5-AS1 reduces ox-LDL-induced damage of HUVECs by targeting miR-106a-5p[J]. Basic & Clinical Medicine, 2022, 42(5): 768-775.

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