MiR-23b-3p regulates lipoprotein (a) to protect human coronary vascular endothelial cells against homocysteine-induced injury

Xiaofeng MA; Huan LIU; Zhaobing LI; Jiankai TAN

doi:10.16016/j.1000-5404.201805205

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MiR-23b-3p regulates lipoprotein (a) to protect human coronary vascular endothelial cells against homocysteine-induced injury

Basic Medicine | 更新时间：2026-01-09

- MiR-23b-3p regulates lipoprotein (a) to protect human coronary vascular endothelial cells against homocysteine-induced injury
- Journal of Third Military Medical University Vol. 40, Issue 22, Pages: 2074-2080(2018)
- 作者机构：
  
  421000 湖南衡阳，南华大学附属南华医院心内科
- 作者简介：
- 基金信息：
  
  the Scientific Research Fund of Health and Family Planning Committee of Hunan Province(B2016139)
- DOI：10.16016/j.1000-5404.201805205
  CLC： R322.12;R394-33;R394.3
- Received：11 May 2018，
  
  Revised：2018-08-01，
  
  Published：30 November 2018
- 稿件说明：
移动端阅览
Xiaofeng MA, Huan LIU, Zhaobing LI, et al. MiR-23b-3p regulates lipoprotein (a) to protect human coronary vascular endothelial cells against homocysteine-induced injury[J]. Journal of Third Military Medical University, 2018, 40(22): 2074-2080.
DOI：

Xiaofeng MA, Huan LIU, Zhaobing LI, et al. MiR-23b-3p regulates lipoprotein (a) to protect human coronary vascular endothelial cells against homocysteine-induced injury[J]. Journal of Third Military Medical University, 2018, 40(22): 2074-2080. DOI： 10.16016/j.1000-5404.201805205.

摘要

目的

探讨miR-23b-3p作为一种高效降低载脂蛋白(a)[apolipoprotein(a)，Apo(a)]的内源性miR，对高同型半胱氨酸引起的冠状动脉内皮细胞损伤抑制效果及作用机制。

方法

通过生物信息学、miR芯片及基础实验筛选ETS1为靶基因，设计miR-23b-3p mimic/inhibitor。构建pmirGLO- ETS1 3′-UTR质粒分6组转染人冠状动脉内皮细胞(human coronary artery endothelial cells，HCAECs)，分别为A组(pmirGLO-ETS1 3′-UTR转染)、B组(miR-23b-3p inhibitor+pmirGLO-ETS1 3′-UTR转染)、C组(miR-23b-3p minics+pmirGLO-ETS13′-UTR转染)、D组(pmirGLO-ETS13′-UTR mut转染)、E组(miR-23b-3p minics+pmirGLO-ETS13′-UTR mut转染)和F组(miR-23b-3p inhibitor +pmirGLO-ETS13′-UTR mut转染)，经双荧光素酶报告基因实验验证。HCAECs分为：对照组(无干预)、阴性对照组(miR-23b-3p inhibitor转染)和实验组(miR-23b-3p mimic转染)。3组细胞均置于含1.0 mmol/L同型半胱氨酸(homocysteine

hcy)培养基中培养7 d。检测各组HCAECs存活率、miR-23b-3p和Apo(a) mRNA表达、akt、p-akt和eNOS蛋白表达。

结果

双荧光素酶报告基因实验验证miR-23b-3p能够和ETS1 3′-UTR直接结合，可得ETS1是miR-23b-3p直接靶基因。实验组HCAECs存活率(66.22±5.17)%、阴性对照组(42.12±2.35)%和对照组(38.36±2.21)%均显著低于正常HCAECs(

0.01)，实验组HCAECs存活率显著高于阴性对照组和对照组(

0.01)。转染后，实验组miR-23b-3p mRNA表达显著高于阴性对照组和对照组(

0.01)，实验组Apo(a) mRNA表达显著低于阴性对照组和对照组(

0.01)。转染后，实验组p-akt表达和eNOS蛋白相对表达显著高于阴性对照组和对照组(

0.01)。

结论

miR-23b-3p能够以ETS1为靶基因，可能通过akt/ eNOS信号通路下调hcy诱导损伤的内皮细胞中Apo(a)表达，从而保护冠状动脉血管内皮细胞。

Abstract

Objective

To explore the role of miR-23b-3p

an endogenous microRNA that efficiently lowers apolipoprotein (a) [Apo (a)] level

in protecting human coronary artery endothelial cells (HCAECs) against homocysteine-induced injury.

Methods

Based on the results of bioinformatics analysis

miR chip data and preliminary experimental findings

ETS1 was selected as the target gene for designing miR-23b-3p mimic and inhibitor. The pmirGLO-ETS1 3′-UTR and pmirGLO-ETS1 3′-UTR mut plasmids were constructed and transfected separately into cultured HCAECs along with the miR-23b-3p inhibitor or miR-23b-3p mimics

followed by testing of the double luciferase reporter gene. HCAECs transfected with the miR-23b-3p mimic

miR-23b-3p inhibitor or without transfection were cultured in the presence of 1.0 mmol/L homocysteine for 7 d

and the cell survival rate

expression of miR-23b-3p and Apo (a) mRNA

and expression of akt

p-akt and eNOS proteins were measured.

Results

Double luciferase reporter gene assay showed a direct binding of miR-23b-3p to ETS1 3′-UTR

suggesting that ETS1 was the direct target gene of miR-23b-3p. After exposure to homocysteine for 7 d

the survival rate of HCAECs cells transfected with the miR-23b-3p mimic was (66.22±5.17)%

significantly higher than the rate of (42.12±2.35)% in cells transfected with the miR-23b-3p inhibitor and the rate of (38.36±2.21)% in cells without transfection (

0.01); the cell survival rates in the 3 groups were all significantly lower than that of normal HCAECs (

0.01). Transfection of HCAECs with the miR-23b-3p mimic significantly up-regulated the expression level of miR-23b-3p mRNA (

0.01)

lowered the expression level of Apo (a) mRNA (

0.01)

and increased the expression levels of p-Akt and eNOS proteins (

0.01).

Conclusion

With ETS1 as the target gene

miR-23b-3p decreases the expression level of Apo (a) through the akt/eNOS signaling pathway to protect HCAECs against homocysteine-induced injury.

关键词

Keywords

references

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