ApoE-/-小鼠动脉粥样硬化斑块中NHE1与Netrin-1表达上调

基础医学与临床 ›› 2020, Vol. 40 ›› Issue (6): 777-783.

ApoE^-/-小鼠动脉粥样硬化斑块中NHE1与Netrin-1表达上调

杨卉¹, 莫显刚^2*, 王兰², 张诗悦¹

1.贵州医科大学临床医学系;
2.贵州医科大学附属医院综合病房, 贵州贵阳 550004

收稿日期:2018-11-04 修回日期:2019-09-09 出版日期:2020-06-05 发布日期:2020-05-29
通讯作者: *moxiangang123@126.com
基金资助:
国家自然科学基金(31660288)

Up-regulated expression of NHE1 and Netrin-1 in atherosclerotic plaque of ApoE^-/-mice

YANG Hui¹, MO Xian-gang^2*, WANG Lan², ZHANG Shi-yue¹

1. Department of Clinical Medicine, Guizhou Medical University;
2. Comprehensive Ward,the Affiliated Hospital of Guizhou Medical University,Guizhou 550004,China

Received:2018-11-04 Revised:2019-09-09 Online:2020-06-05 Published:2020-05-29
Contact: *moxiangang123@126.com

摘要/Abstract

摘要： 目的探讨ApoE^-/-小鼠动脉粥样硬化斑块组织及小鼠巨噬细胞系(RAW 264.7)是否同时表达钠氢交换体1(NHE1)及神经轴突导向因子-1(netrin-1),以及两者之间共定位、相互影响情况。方法制作小鼠动脉粥样硬化斑块模型,HE染色检测主动脉粥样斑块形成;免疫荧光及显微共聚焦检测动脉粥样斑块中NHE1、netrin-1表达及共定位;用氧化型低密度脂蛋白(Ox-LDL)干预RAW264.7细胞,Western blot检测不同浓度及时间干预下, NHE1和netrin-1蛋白表达;设计NHE1基因小干扰RNA(siRNA),通过脂质体转染导入RAW264.7细胞,RT-qPCR及Western blot分别检测靶向作用于NHE1基因的小干扰RNA(siRNA)的抑制效率。结果小鼠动脉硬化斑块中NHE1表达上调(P＜0.05),且NHE1与netrin-1存在部分共定位情况;Ox-LDL可上调RAW264.7细胞NHE1及netrin-1蛋白表达,且呈一定时间-浓度依赖性(P＜0.05);构建的NHE1 siRNA能有效抑制RAW264.7细胞NHE1表达(P＜0.05);有效敲降RAW264.7细胞NHE1表达能使Ox-LDL干预下升高的NHE1、netrin-1蛋白表达下调(P＜0.05)。结论小鼠动脉粥样硬化斑块及Ox-LDL干预的小鼠巨噬细胞系(RAW264.7)中NHE1、netrin-1表达上调且存在共定位规律,netrin-1表达受NHE1表达的影响。

关键词: 动脉粥样硬化, 钠氢交换体1, 神经轴突导向因子1

Abstract: Objective To investigate whether ApoE^-/- mouse atherosclerotic plaque tissue and RAW 264.7 simultaneously express sodium hydrogen exchanger 1 (NHE1) and neurite guidance factor-1 (netrin-1), and their co-localization and interaction. Methods The atherosclerotic plaque model of mice was established, and the formation of atherosclerotic plaque was detected by HE staining; the expression and co-localization of NHE1 and netrin-1 in atherosclerotic plaque were detected by immunofluorescence and confocal microscopy; Oxidized low density lipoprotein (Ox-LDL) was used to interfere with RAW264.7 cells in mice, then Western blot assay was used to detect the expression of NHE 1 and netrin-1 proteins in samples with different concentrations and time intervention. Small interfering RNA (siRNA) targeting NHE1 gene was designed and transfected into RAW264.7 cells by liposome. The inhibition efficiency of small interfering RNA (siRNA) targeting NHE1 gene was detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and Western blot. Results The expression of NHE1 was up-regulated in atherosclerotic plaque of mice, and partly co-localized with netrin-1. Ox-LDL up-regulated the expression of NHE1 and netrin-1 in RAW264.7 cells in a time-concentration dependent manner (P＜0.05); the constructed NHE1 siRNA effectively inhibited the expression of NHE1 in RAW264.7 cells (P＜0.05); effective knockdown of NHE1 expression in RAW264.7 cells down-regulated the expression of increased NHE1 and netrin-1 proteins as a results of intervention by Ox-LDL(P＜0.05). Conclusions The expression of NHE1 and netrin-1 in atherosclerotic plaque and Ox-LDL-treated mouse macrophage line (RAW264.7) is up-regulated and co-localized. The expression of netrin-1 is affected by the expression of NHE1.

Key words: atherosclerosis, sodium hydrogen exchangers 1, netrin-1

中图分类号:

R332

杨卉, 莫显刚, 王兰, 张诗悦. ApoE^-/-小鼠动脉粥样硬化斑块中NHE1与Netrin-1表达上调[J]. 基础医学与临床, 2020, 40(6): 777-783.

YANG Hui, MO Xian-gang, WANG Lan, ZHANG Shi-yue. Up-regulated expression of NHE1 and Netrin-1 in atherosclerotic plaque of ApoE^-/-mice[J]. Basic & Clinical Medicine, 2020, 40(6): 777-783.

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